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See detailMethod for the production of recombinant proteins by green microalgae
Versali, Marie-France ULg; Clerisse, Fabienne ULg; Dommes, Jacques ULg et al

Patent (2011)

The present invention relates to a method for the production of recombinant proteins by algal cells, which are fresh water unicellular green microalgae belonging to the order of Sphaeropleales. The ... [more ▼]

The present invention relates to a method for the production of recombinant proteins by algal cells, which are fresh water unicellular green microalgae belonging to the order of Sphaeropleales. The present invention further provides recombinant algal cells, wherein said algal cells are fresh water unicellular green microalgae belonging to the order of Sphaeropleales, and wherein said algal cells are capable of producing a recombinant protein. The invention is also directed to a method for selecting recombinant algal cells. The invention also relates to the use of recombinant algal cells as described herein for producing recombinant proteins. [less ▲]

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See detailDo polyamines increase the antioxidant capacity of hyperhydric shoots ?
Franck, Thierry ULg; Tabart, Jessica ULg; Kevers, Claire ULg et al

in 4th international workshop - cost action FA0605 - Book of abstracts - Limassol 17-19 November (2011)

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See detailAscorbic acid, Phenolic acid, Flavonoid and Carotenoid profiles of selected extracts from Ribes nigrum
Tabart, Jessica ULg; Kevers, Claire ULg; Evers, Danièle et al

in Journal of Agricultural and Food Chemistry (2011), 59

Small fruits such as berries have low energy contents, but high content of vitamins, micronutrients, and dietary fibres and constitute a good source of natural antioxidant compounds that are important ... [more ▼]

Small fruits such as berries have low energy contents, but high content of vitamins, micronutrients, and dietary fibres and constitute a good source of natural antioxidant compounds that are important constituents of the human diet. This study allowed to identify a large number of compounds in an extract of blackcurrant showing high antioxidant activity and to compare their profile in various parts of the plants (leaves, buds and berries). If it was known that berries contained very high levels of natural phenolic compounds, here we showed that leaves and buds could also be considered as a good sources of natural antioxidants. Indeed, they contained high amount of ascorbic acid, phenolic acids, flavonoids and carotenoids. An acetone mixture can extract several classes of phenolic compounds with a good yield of ascorbic acid, flavonols, flavan-3-ols and anthocyanins. For phenolic acids, it extracted fewer compounds than a specific extraction solution. [less ▲]

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See detailInfluence of cultivar, harvest time, storage conditions, and peeling on the antioxidant capacity and phenolic and ascorbic acid contents of apples and pears."
Kevers, Claire ULg; PINCEMAIL, Joël ULg; Tabart, Jessica ULg et al

in Journal of Agricultural and Food Chemistry (2011), 59

Apple and pear fruits are important sources of secondary plant metabolites and one of the major sources of dietary phenolics consumed all year round. The aim of this work was to identify the main ... [more ▼]

Apple and pear fruits are important sources of secondary plant metabolites and one of the major sources of dietary phenolics consumed all year round. The aim of this work was to identify the main variables influencing phenolic content and antioxidant capacity in apples. Higher phenolic and antioxidant contents were observed in some varieties (such as the Delbar Estival apple and Durondeau pear). Storage conditions were important. Our results also showed that fruits should be consumed rapidly after purchase and with their peel. After one week of domestic storage, the ascorbic acid content was found to decrease by 75%. Peeling led to a more than 25% decrease in total phenolics and ascorbic acid. The harvest time (at normal ripeness) had only a limited impact, but significant year-to-year variations were observed. In conclusion, well-chosen and well-stored apples and pears may contribute to an antioxidant-rich diet if consumed rapidly and with their peel. [less ▲]

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See detailThe elicitation of a systemic resistance by Pseudomonas putida BTP1 in tomato involves the stimulation of two lipoxygenase isoforms
Mariutto, Martin ULg; Duby, Franceline ULg; Adam, Akram et al

in BMC Plant Biology (2011), 11

Background Some non-pathogenic rhizobacteria called Plant Growth Promoting Rhizobacteria (PGPR) possess the capacity to induce in plant defense mechanisms effective against pathogens. Precedent studies ... [more ▼]

Background Some non-pathogenic rhizobacteria called Plant Growth Promoting Rhizobacteria (PGPR) possess the capacity to induce in plant defense mechanisms effective against pathogens. Precedent studies showed the ability of Pseudomonas putida BTP1 to induce PGPR-mediated resistance, termed ISR (Induced Systemic Resistance), in different plant species. Despite extensive works, molecular defense mechanisms involved in ISR are less well understood that in the case of pathogen induced systemic acquired resistance. Results We analyzed the activities of phenylalanine ammonia-lyase (PAL) and lipoxygenase (LOX), key enzymes of the phenylpropanoid and oxylipin pathways respectively, in tomato treated or not with P. putida BTP1. The bacterial treatment did not stimulate PAL activity and linoleate-consuming LOX activities. Linolenate-consuming LOX activity, on the contrary, was significantly stimulated in P. putida BTP1-inoculated plants before and two days after infection by B. cinerea. This stimulation is due to the increase of transcription level of two isoforms of LOX: TomLoxD and TomLoxF, a newly identified LOX gene. We showed that recombinant TomLOXF preferentially consumes linolenic acid and produces 13-derivative of fatty acids. After challenging with B. cinerea, the increase of transcription of these two LOX genes and higher linolenic acid-consuming LOX activity were associated with a more rapid accumulation of free 13-hydroperoxy-octadecatrienoic and 13-hydroxy-octadecatrienoic acids, two antifungal oxylipins, in bacterized plants. Conclusion In addition to the discovery of a new LOX gene in tomato, this work is the first to show differential induction of LOX isozymes and a more rapid accumulation of 13-hydroperoxy-octadecatrienoic and 13-hydroxy-octadecatrienoic acids in rhizobacteria mediated-induced systemic resistance. [less ▲]

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See detailIdentification, characterization and expression profiling of the tomato gene TomLoxF
Mariutto, Martin ULg; Duby, Franceline ULg; Adam, Akram et al

Poster (2010, January 26)

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See detailTuber formation and growth of Dioscorea cayenensis-D. rotundata complex. Interactions between exogenous and endogenous jasmonic acid and polyamines
Ondo Ovono, Paul ULg; Kevers, Claire ULg; Dommes, Jacques ULg

in Plant Growth Regulation (2010)

Tubers can be initiated and developed in vitro from nodal cuttings of yam (Dioscorea cayenensis - D. rotundata complex). The effect of exogenous jasmonic acid, alone or in combination with putrescine, on ... [more ▼]

Tubers can be initiated and developed in vitro from nodal cuttings of yam (Dioscorea cayenensis - D. rotundata complex). The effect of exogenous jasmonic acid, alone or in combination with putrescine, on these processes was investigated in relationship to endogenous jasmonic acid and polyamine levels. Application of exogenous jasmonic acid at various concentrations positively affected microtuber formation and growth from yam nodal cuttings. In control conditions, three weeks were needed to obtain 100% of tuberisation. Jasmonic acid at low level (0.1 µM) accelerated tuber formation (46% after one week) as did putrescine (10 µM). But endogenous levels of jasmonic acid were not significantly affected by its exogenous presence in the medium. Jasmonic acid also interacted with other growth regulators as polyamines, but the decrease in time necessary to observe tuber formation could not be correlated with endogenous modifications of PUT content. The presence of jasmonic acid (0.1 to 1 µM) as PUT (1 µM) induced also an increase of tuber length and weight. The combination of jasmonic acid (0.1 µM) and putrescine (1 µM) had no positive effect on tuber formation (precocity) but had an additive effect on further growth (length and weight). In the future, these results could help the optimising in vitro conditions for mass production of larger yam microtubers. [less ▲]

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See detailIn Vitro Preservation of Yam ( Dioscorea cayenensis-D.rotundata complex) for a Better Use of Genetic Resources
Ondo Ovono, Paul ULg; Kevers, Claire ULg; Dommes, Jacques ULg

in Notulae Botanicae Horti Agrobotanici Cluj-Napoca (2010), 38

Among the food crops, yam takes up quantitatively the first place in the gabonese diet. Unfortunately, it can stay available only 6 to 7 months in the year because of difficulties of harvest and post ... [more ▼]

Among the food crops, yam takes up quantitatively the first place in the gabonese diet. Unfortunately, it can stay available only 6 to 7 months in the year because of difficulties of harvest and post- harvest. This problem is little studied in the case of Dioscorea cayenensis-D. rotundata complex. In order to optimize the use of micro tubers for the growing in green house or field, it is important to control the duration of storage before the germination. The present study concerns microtubers obtained by in vitro culture. When microtubers were harvested (after 9 months of culture) and directly transferred on a new medium without hormone, the tubers rapidly sprouted in in vitro conditions. Harvested microtubers were also stored dry in jars in sterile conditions during 2 to 18 weeks before in vitro sprouting. In this case, microtubers stored during 18 weeks sprouted more rapidly than those stored 8 weeks. The size of the tubers used for the storage had great influence on further sprouting. The upper microtubers in 25 mm can be kept to the darkness, under 50% of relative humidity, in 25°C during at least 18 weeks. Sprouting is 100% whatever the substrate of culture. The plant tissue culture technique constitutes a serious alternative for the preservation of plant kinds and for the production of planting material. These techniques allow multiplying in a short time of thousands of copies of new varieties of newly created plants. These in vitro plants can be used on one hand, for the production planting material, and on the other hand for ex vitro storage of breeding grounds with decelerated growth, to struggle against genetic erosion. These results should allow improving in practice the multiplication of yam, while guaranteeing phytosanitary qualities. [less ▲]

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See detailEvaluation of spectrophotometric methods for antioxidant compound measurement in relation to total antioxidant capacity in beverages.
Tabart, Jessica ULg; Kevers, Claire ULg; Pincemail, Joël ULg et al

in Food Chemistry (2010), 120

The validity of different colorimetric methods used to quantify various families of antioxidant compounds was evaluated with standard compounds. The colorimetric tests for global evaluation of flavonoids ... [more ▼]

The validity of different colorimetric methods used to quantify various families of antioxidant compounds was evaluated with standard compounds. The colorimetric tests for global evaluation of flavonoids, anthocyans, and flavanols were found generally unreliable, as reactions could be different for individual compounds within a family (anthocyanins or flavonols or flavan-3-ols) and not specific to one family. In the flavonoid test, for example, flavonols reacted very well, anthocyanins did not react, and flavanons reacted only slightly. The same methods were applied also to beverages known for their antioxidant content (apple, orange, grape, and vegetable juices, ice tea, and red wine) and the data were compared with the results of HPLC analysis of specific compounds. The values obtained in a colorimetric test were generally higher than the sum of the values obtained for the corresponding individual compounds by HPLC analysis, mainly because other compounds can interfere with the colorimetric tests. For example, in wine, anthocyanin concentrations obtained by colorimetric test were 9068  1407 µ mol /100ml (mean  SEM), higher than the sum of the six main anthocyanidins detected by HPLC, only 41 µmol/100 ml. The relative antioxidant capacity values determined for beverages on the basis of colorimetric tests could exceed by far the values previously measured in radical-scavenging tests (for instance, the antioxidant capacity attributable to anthocyans in wine on the basis of the colorimetric test was 50 times higher than the total antioxidant capacity measured by the ORAC assay). In conclusion, colorimetric tests for flavonoids, anthocyans, and flavanols appeared generally unreliable for estimating their content and thus the antioxidant capacity reliable to these compounds. [less ▲]

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See detailEffects of storage conditions on sprouting of microtubers of yam (Dioscorea cayenensis-D. rotundata complex).
Ondo Ovono, Paul ULg; Kevers, Claire ULg; Dommes, Jacques ULg

in Comptes Rendus des Séances de l'Académie des Sciences. Série III, Sciences de la Vie (2010)

The control of field tuber dormancy in yam (Dioscorea cayenensis- D.rotundata complex) is poorly understood. Although studies have examined single environmental factors and chemical treatments that might ... [more ▼]

The control of field tuber dormancy in yam (Dioscorea cayenensis- D.rotundata complex) is poorly understood. Although studies have examined single environmental factors and chemical treatments that might prolong tuber dormancy and storage, only a few were focused on further tuber sprouting. The present study concerns microtubers obtained by in vitro culture. When microtubers were harvested (after 9 months of culture) and directly transferred on a new medium without hormone, the tubers rapidly sprouted in in vitro conditions. No dormancy was observed in this case. Harvested microtubers were also stored dry in jars in sterile conditions during 2 to 18 weeks before in vitro sprouting. In this case, microtubers stored during 18 weeks sprouted more rapidly than those stored 8 weeks. A constant “dormancy-like period” (storage duration + sprouting delay) was observed, between 20 and 28 weeks respectively for the more rapid and the slower microtubers. The size of the tubers used for the storage had great influence on further sprouting. The larger they were, the better they sprouted. Light during storage had no effect on the sprouting delay while a temperature of 25 °C permit a quicker sprouting than 18 °C. The medium used to obtain microtubers could also have an effect on sprouting rate. Ex vitro sprouting was not a problem. There was a delay in sprouting in contrast to in vitro conditions but the rate of 100% was kept. This fact was very important for an agronomical application of this technique to the production of “seeds” directly usable in the field or after culture in greenhouse. [less ▲]

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See detailANTIOXIDANT CAPACITY OF THREE CUBAN SPECIES OF THE GENUS Pluchea Cass. (Asteraceae)
Perera, Wilmer; Tabart, Jessica ULg; Gomez, Abel et al

in Journal of Food Biochemistry (2010), 34

Leaves of three Cuban species of the genus Pluchea: P. carolinensis, P. odorata and P. rosea were extracted with various solvents and analysed. Highest values of phenolic compounds were detected after ... [more ▼]

Leaves of three Cuban species of the genus Pluchea: P. carolinensis, P. odorata and P. rosea were extracted with various solvents and analysed. Highest values of phenolic compounds were detected after ethyl acetate (EtOAc) and n-butyl alcohol (n-BuOH) extractions. Flavonoids were detected after chloroform, EtOAc and n-BuOH extractions of the species Pluchea and three aglycone flavonol forms (quercetin, kaempferol and myricetin) were also quantified after EtOAc and n-BuOH extractions, using HPLC. The total antioxidant capacity (TAC), expressed as Trolox equivalents per gram of leaf dry weight (TE/g dw) was analyzed using 2,2-diphenyl-1-picrylhydrazyl (DPPH•), 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS∙+), and the oxygen radical absorbance capacity (ORAC). The highest values of TAC were detected after EtOAc and n-BuOH extractions of the three species. P. carolinensis was the promising species; being the n-BuOH extraction with the most bioactive compounds : 15.3 mg TE/g dw using DPPH∙ [less ▲]

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See detailTuber formation and development of Dioscorea cayenensis-D. rotundata complex in vitro. Effect of polyamines.
Ondo Ovono, Paul ULg; Kevers, Claire ULg; Dommes, Jacques ULg

in In Vitro Cellular & Developmental Biology-Plant (2009)

Tuberisation was obtained in vitro on yam (Dioscorea cayenensis-D.rotundata complex). The effect of exogenous polyamines on tuber formation and development (length and weight of microtubers) were ... [more ▼]

Tuberisation was obtained in vitro on yam (Dioscorea cayenensis-D.rotundata complex). The effect of exogenous polyamines on tuber formation and development (length and weight of microtubers) were investigated and discussed in relation with changes in endogenous polyamines. Application of exogenous polyamines, inhibitors of their metabolism and polyamines precursors in various concentrations positively affected microtuber formation by yam nodal cuttings and their further development. In control conditions, three weeks are needed to obtain 100% of tuberisation. With low concentrations of putrescine (10 5 or 10 6 M), tuber formation occurred earlier. Polyamine endogenous level and metabolism can be significantly affected by exogenous polyamines but modifications of endogenous free polyamines could not be directly correlated to the tuber formation process. Increases in endogenous putrescine and auxins were observed in tubers showing a better development in the presence of putrescine. These results can be used for optimising in vitro conditions for mass production of larger microtubers of the Dioscorea cayenensis-D. rotundata complex. [less ▲]

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See detailEffects of reducing sugar concentration on in vitro tuber formation and sprouting in yam (Dioscorea cayenensis-D. rotundata complex).
Ondo Ovono, Paul ULg; Kevers, Claire ULg; Dommes, Jacques ULg

in Plant Cell, Tissue & Organ Culture (2009), 99(1), 55-59

The effects of reducing sucrose level on tuber formation (% of cultures with microtubers), development (length and fresh weight of microtubers) and sprouting in yam Dioscorea cayenensis-D. rotundata ... [more ▼]

The effects of reducing sucrose level on tuber formation (% of cultures with microtubers), development (length and fresh weight of microtubers) and sprouting in yam Dioscorea cayenensis-D. rotundata complex in vitro were investigated. Only 29% of the explants showed tuber formation after 3 weeks in the presence of 1% sucrose in contrast to 100% with 3%. After 120 days of culture, the length and the weight of the tubers obtained in the presence of 1% sucrose were less than with 3% sucrose. Addition of sorbitol to keep osmolarity at the same level did not restore normal rate of tuber formation. Similar results were obtained with the use of reduced fructose or glucose level. Microtuber sprouting was also affected by sucrose level incorporated into the tuberisation medium. Tubers obtained on reduced sucrose level sprouted later and the increase of osmolarity with sorbitol did not restore normal sprouting. The bigger tubers obtained on high sucrose media could contain more carbohydrate reserves that could partially explain a higher sprouting rate. These results can be used for optimising in vitro conditions for mass production of microtubers in yam and especially in Dioscorea cayenensis-D. rotundata complex, a very important species in West Africa. They specially showed the importance of tuberisation conditions on precocity of tuberisation, on tuber length and weight and on their further sprouting. [less ▲]

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See detailInsights into the Defense-Related Events Occuring in Plant Cells Following Perception of Surfactin-Type Lipopeptide from Bacillus subtilis
Jourdan, Emmanuel ULg; Henry, Guillaume; Duby, F. et al

in Molecular Plant-Microbe Interactions [=MPMI] (2009), 22(4), 456-468

Multiple strains of Bacillus subtilis were demonstrated to stimulate plant defense responses, and cyclic lipopeptides may be involved in the elicitation of this induced systemic resistance phenomenon ... [more ▼]

Multiple strains of Bacillus subtilis were demonstrated to stimulate plant defense responses, and cyclic lipopeptides may be involved in the elicitation of this induced systemic resistance phenomenon. Here, we further investigated molecular events underlying the interaction between wuch lipopeptides and plant cells. Addition of surfactin but not fengycin or iturin in the micromolar range to tobacco cell suspensions induced defense-related early events such as extracellular medium alkalinization coupled with ion fluxes and reactive oxygen species production. Surfactin also stimulated the defense enzymes phenylalanine ammonia lyase and lipoxygenase and modified the pattern of phenolics produced by the elicited cells. The occurence of these surfactin-elicited early events is closely related to Ca2+ influx and dynamic changes in protein phosphorylation but is not associed with any marked phytotoxicity or adverse effect on the integrity and growth potential of the treated tobacco cells. [less ▲]

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See detailCombining proteomics and metabolite analyses to unravel cadmium stress-response in poplar leaves
Kieffer, Pol ULg; Planchon, Sébastien; Oufir, Mouhssin et al

in Journal of Proteome Research (2009), 8

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See detailWhat we have learned about the physiology of in vitro adventitious rooting of woody plants and how it relates to improvements in the practice
Kevers, Claire ULg; Hausman, Jean-françois; Faivre-Rampant, Odile et al

in Niemi, Karoliina (Ed.) Adventitious root formation of forest trees and horticultural plants - from genes to applications (2009)

Natural auxins and synthetic analogs are the most powerful exogenous stimulators of adventitious rooting of ligneous as well as for herbaceous cuttings. For several years we have investigated the ... [more ▼]

Natural auxins and synthetic analogs are the most powerful exogenous stimulators of adventitious rooting of ligneous as well as for herbaceous cuttings. For several years we have investigated the physiology of rooting, including the metabolism of endogenous auxins and their molecular mechanisms of action. Our work has resulted in discoveries leading to practical strategies allowing faster and improved rooting of cuttings from shoots raised in vitro, and subsequently resulting in roots better adapted for acclimatization and to overcome so-called (apparent) recalcitrance-to-rooting of some species. In this chapter we describe: (i) how variation in auxin treatments during rooting can be used to regulate the endogenous levels of auxins; (ii) the importance of not maintaining auxin treatment throughout the whole rooting process, thereby the endogenous auxin level itself not always being high; (iii) the division of the organogenetic process into at least three main rooting interdependent phases, i.e. induction, initiation, and expression, with different durations depending on the species and with different requirements for each phase, and (iv) how polyamines, even if less effective from outside, and their metabolism are as important as auxins in the rooting process. We will also describe how cross-talk between hormones, manipulations with related genes and mutants, and some similarities between the induction of rooting and evocation of flowering, offer new tools for further progress in the field. [less ▲]

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See detailInvolvement of polyamines in the adventitious rooting of micropropagated shoots of the apple rootstock MM106
Naija, Sélima; Elloumi, Nadhra; Ammar, Saida et al

in In Vitro Cellular & Developmental Biology-Plant (2009), 45

Apple rootstock MM106 shoots, raised in vitro, rooted at 96.7% after culture on a medium supplemented with an auxin for 5 d in darkness followed by culture on a second medium without growth regulators for ... [more ▼]

Apple rootstock MM106 shoots, raised in vitro, rooted at 96.7% after culture on a medium supplemented with an auxin for 5 d in darkness followed by culture on a second medium without growth regulators for 25 d in light. In control conditions (in absence of auxin in the first medium), these shoots did not root. Putrescine (PUT), spermidine (SPD), cyclohexylamine (CHA), and aminoguanidine (AG) enhanced rooting when applied during the first d of culture in the absence of IBA; on the contrary, α-difluoromethylornithine (DFMO) added to the first medium with IBA inhibited rooting. The endogenous levels of indole 3-acetic acid (IAA) and indole 3-acetylaspartic acid (IAAsp) increased up to a maximum concentration at days 2 and 3, respectively, in initial rooting conditions. PUT, when added with IBA, did not affect the typical IAA and IAAsp increase; when applied alone, it provoked an increase of their levels. Similar results were recorded with CHA. SPD, AG, and DFMO did not induce an increase of IAA and IAAsp in nonrooting conditions. The levels of endogenous PUT increased to a maximum at day 2 in rooting conditions; it was slightly affected by exogenous PUT and CHA application but reduced by SPD, AG, and DFMO. In rooting conditions, if the first medium was supplemented with SPD or AG, a small increase in peroxidase activity was observed, similar to that obtained with PUT treatment. The present work indicates an involvement of polyamines in the control of rooting and an interaction with auxins during the physiological phase of rooting. The consequence of this relationship was a different rooting expression, according especially to the content of these regulators in the culture medium. [less ▲]

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See detailComparative antioxidant capacities of phenolic compounds measured by various tests
Tabart, Jessica ULg; Kevers, Claire ULg; Pincemail, Joël ULg et al

in Food Chemistry (2009), 113

The purpose of this study was to compare the antioxidant capacities of standard compounds (phenolic compounds, ascorbic acid, and glutathione) as measured by various assays. Five methods were selected so ... [more ▼]

The purpose of this study was to compare the antioxidant capacities of standard compounds (phenolic compounds, ascorbic acid, and glutathione) as measured by various assays. Five methods were selected so as to span a diversity of technical approaches: TEAC (radical 2,2′-azino-bis(3-ethylbenzothiazoline)-6 sulphonic acid), DPPH (radical 2,2-diphenyl-1-picryhydrazyl used to measure reducing capacity), ORAC (oxygen radical scavenging capacity), red blood cell haemolysis (protection of biological sample), and ESR (electron spin resonance for direct free radical evaluation). Most compounds showed significant differences in free radical scavenging activity according to the method used. Of the 25 tested compounds, only a few, such as myricetin and gallocatechin, gave comparable activities in the various tests. To standardise reporting on antioxidant capacity, it is proposed to use a weighted mean of the values obtained using the DPPH, ORAC, resistance to haemolysis, and ESR assays. This strategy was used to test the antioxidant capacity of several beverages. The highest antioxidant capacity was observed for red wine, followed by green tea, orange juice, grape juice, vegetable juice, and apple juice. [less ▲]

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