<Emphasis Type="Italic">In vitro</Emphasis> and <Emphasis Type="Italic">In vivo</Emphasis> Antioxidant Properties of Extracts from <Emphasis Type="Italic">Coptis chinensis</Emphasis> Inflorescence

The inflorescence of cultivated Coptis chinensis has been valued for tea production for many years in China. The antioxidant activities of C. chinensis inflorescence extracts prepared by various solvents were investigated by using several established in vitro systems: 2,2′-azinobis(3-ethylbenzthiazoline-6-sulphonic acid (ABTS), α,α-diphenyl-β-picrylhydrazyl (DPPH) and superoxide radical scavenging assays, reducing power assay, and ferrothiocyanate (FTC) and thiobarbituric acid (TBA) assays. The results showed that the 70% ethanol extract (EE) had the strongest antioxidant activity in vitro among the various extracts. Based on the in vitro results, EE was used to evaluate the antioxidant activity of C. chinensis inflorescence in vivo. The liver and kidney of intoxicated animals showed a significant decrease in superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) levels, while the malondialdehyde (MDA) level showed a significant increase. These changes were significantly reversed after treatment with EE and the standard vitamin E. Thus, the C. chinensis inflorescence may be a valuable natural source that can be applicable to food industries.     

The Low Potential of Teff (<Emphasis Type="Italic">Eragrostis tef</Emphasis>) as an Inoculum Source for <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Teff (Eragrostis tef) is a fine stemmed annual grass and gluten free small grain that is of interest as a forage, cover, or a rotation crop. Little is known about the susceptibility of teff to many diseases. Teff could be grown in rotation with potato in the northwestern United States provided teff cultivation is economical and does not increase soil populations for pathogens affecting rotation crops such as Verticillium dahliae. Verticillium dahliae infects a wide range of dicotyledonous plants, making it one of the most important fungal pathogens of crop plants in North America, including potato. The objective of this study was to quantify the susceptibility of teff to eight V. dahliae isolates and compare the susceptibility of teff to eggplant. Teff was confirmed as a host for V. dahliae, as indicated by the presence of microsclerotia in teff stems and roots after artificial inoculation in two years of greenhouse studies. The number of microsclerotia produced in teff did not differ between mint and potato pathotypes of V. dahliae. No V. dahliae isolate produced significantly greater numbers of microsclerotia than any of the seven other isolates tested in a two-year study. Microsclerotia production of V. dahliae in teff was consistently less than in susceptible eggplant cv. Night shadow in both greenhouse experiments (P?<?0.02). It is unlikely that teff infected by V. dahliae will proliferate microsclerotia of mint or potato-aggressive pathotypes, especially when compared to susceptible eggplant cultivars.     

Bioactivity Improvement of <Emphasis Type="Italic">Olea europaea</Emphasis> Leaf Extract Biotransformed by <Emphasis Type="Italic">Wickerhamomyces anomalus</Emphasis> Enzymes

Olive leaves represent a quantitatively significant by-product of agroindustry. They are rich in phenols, mainly oleuropein, which can be hydrolyzed into several bioactive compounds, including hydroxytyrosol. In this study, water extract from olive leaves ‘Biancolilla’ was analyzed for polyphenol profile, DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity and protective effect on differentiated Caco-2 cells. The efficacy of two enzymatic treatments in promoting the release of bioactive phenols was investigated: a) enzymatic extract from Wickerhamomyces anomalus, characterized by β-glucosidase and esterase activities; b) commercial β-glucosidase. Composition and bioactivity of the resulting extracts were compared. The results showed that the yeast-treated extract presented hydroxytyrosol content and DPPH radical scavenging activity comparable to those obtained using commercial β-glucosidase; however, it was showed the additional presence of hydroxycinnamic acids. In experiments on Caco-2 cells, the leaf extracts promoted the recovery of cell membrane barrier at different minimum effective concentrations. The high specificity of W. anomalus enzymatic extract may represent an effective tool for the release of bioactive phenols from olive by-products.     

<Emphasis Type="Italic">In vitro</Emphasis> Antioxidant Activity of Aged Extracts of some Italian <Emphasis Type="Italic">Allium</Emphasis> Species

Antioxidant activity of fresh Allium sativum L. (garlic) is well known and is mainly due to unstable and irritating organosulphur compounds. Fresh garlic extracted over a prolonged period (up to 20 months) produces odourless aged garlic extract (AGE) containing stable and water soluble organosulphur compounds that prevent oxidative damage by scavenging free radicals. The aim of this study was to investigate the in vitro antioxidant activity of aged (up to 20 months) 15% hydroethanolic extracts of different parts (bulbs, bulblets, flower bulblets, flowers, and leaves) of three Allium spontaneous species which are endemic for Italian flora: Allium neapolitanum Cyr., Allium subhirsutum L., Allium roseum L. and to compare it with the in vitro antioxidant activity of aged 15% hydroethanolic extracts of bulbs and leaves of garlic. The antioxidant potential of aged extracts of all species has been evaluated using two different spectrophotometric assays: 2,2-diphenylpicrylhydrazyl (DPPH) test and the ferric reducing/antioxidant power (FRAP) assay. Furthermore the polyphenol content was determined. The aged extracts obtained from the leaves showed the best antioxidant activity, followed by flowers and then by bulbs in both used tests, while flower bulblets and bulblets exhibited lower results or no activity. The polyphenol content was generally directly correlated with antioxidant/antiradical activity. This study confirms the data obtained in previous researches, the wild-type species of Allium and in particular organs other than bulbs are more active and efficacious than garlic bulb. Surely leaves of these Allium spp. deserve special attention.     

Phenolic Composition,Antioxidant Capacity and <Emphasis Type="Italic">In Vitro</Emphasis> Cancer Cell Cytotoxicity of Nine Prickly Pear (<Emphasis Type="Italic">Opuntia</Emphasis> spp.) Juices

Juices of nine prickly pears (Opuntia spp.) were characterized in terms of color, acidity, sugar content, phenolics, flavonoids, betalains and antioxidant activity and tested in vitro against four cancer cell lines. The juices had pH′s, acidities and sugar ranging from 4.27 to 5.46, 0.03 to 0.27% and 8 to 14.7°Brix, respectively. Juices also varied in color from white to purple and contained total phenolics, flavonoids, betaxanthins, betacyanins and antioxidant capacity ranging from 22 to 226 μg gallic acid eq/g, 95 to 374 μg quercetin eq/g, 3 to 189 μg/g, 1.6 to 300 μg/g and 17 to 25 micromoles Trolox eq./mL, respectively. Among the cancer lines tested, viability of prostate and colon cells were the most affected. Moradillo contained the highest flavonoids and diminished both prostate and colon cancer cell viability without affecting mammary or hepatic cancer cells. Rastrero reduced the growth of the four cancer cell lines without affecting normal fibroblast viability. The research shows intervarietal differences among prickly pears in terms of juice properties and phytochemicals that could prevent oxidative stress and cancer.     

Polyphenolic Composition and Antioxidant Characteristics of Kumquat (<Emphasis Type="Italic">Fortunella margarita</Emphasis>) Peel Fractions

The polyphenolic composition of two Fortunella margarita (Nagami kumquat) specimens from Greece and Egypt was investigated employing fractionation by solvent partition and liquid chromatography-mass spectrometry. The main groups of phenolics identified in the different fractions generated were C-glycosylated flavones, O-glycosylated flavones, C-glycosylated flavanones, O-glycosylated flavanones, flavonols, chalcones, phenolic acids and derivatives thereof. The antioxidant potency of the fractions was assessed using two representative in vitro tests, including antiradical activity and hydroxyl free radical scavenging activity. It was revealed that the ethyl acetate fractions from both specimens contained the higher polyphenol content and exhibited the better antioxidant characteristics. The results indicated that F. margarita peels may be regarded as a rich source of potentially bioactive polyphenols.     

<Emphasis Type="Italic">Prunus persica</Emphasis> var. <Emphasis Type="Italic">platycarpa</Emphasis> (Tabacchiera Peach): Bioactive Compounds and Antioxidant Activity of Pulp,Peel and Seed Ethanolic Extracts

A comparative analysis of ethanol extracts from peel, pulp and seed of Prunus persica var. platycarpa (Tabacchiera peach) was done. The total phenol, flavonoid and carotenoid content as well as the antioxidant properties by using different in vitro assays (DPPH, ABTS, FRAP, Fe-chelating, β-carotene bleaching test) were evaluated. Pulp extract was subjected to liquid chromatography-electrospray-tandem mass spectrometry (HPLC-ESI-MS/MS). Gallic acid, protocatechuic acid, protocatechualdehyde, chlorogenic acid, p-coumaric acid, and ferulic acid were identified as main constituents. Pulp extract was characterized by the highest total phytonutrients content and exhibited the highest antioxidant activity in all in vitro assays (IC₅₀ values of 2.2 μg/mL after 60 min of incubation by using β-carotene bleaching test and 2.9 μg/mL by using Fe-chelating assay). Overall, the obtained results suggest that P. persica var. platycarpa displays a good antioxidant activity and its consumption could be promoted.     

The Antioxidant and Anti-proliferative Activity of the Lebanese <Emphasis Type="Italic">Olea europaea</Emphasis> Extract

It is becoming increasingly evident that certain phytochemicals possess cancer chemopreventive properties. In this study, the anti-proliferative activity of plant extracts from olive (Olea europaea L.) leaves was tested on human leukemic cell line (Jurkat). Cytotoxicity of various concentrations of plant extracts was examined and the IC₅₀ was determined. Olive leaf extracts showed concentration-dependent anti-proliferative effect as determined by the WST-1 proliferation kit and [³H]-thymidine incorporation method. To study whether cell death was due to apoptosis, cells were stained with Annexin V-FITC and PI and the expression of important regulatory proteins (Bcl-2, Bax, and p53) involved in apoptosis were examined by Western blot. The antioxidant activity of olive leaves (SC₅₀ = 0.1 mg dry weight) was studied using the DPPH scavenging method. Present findings suggest that olive leaves extracts exhibit anti-proliferative effect on leukemic cells by inducing apoptosis.     

Comparative Study on the Chemical Composition,Antioxidant Properties and Hypoglycaemic Activities of Two <Emphasis Type="Italic">Capsicum annuum</Emphasis> L. Cultivars (<Emphasis Type="Italic">Acuminatum</Emphasis> small and <Emphasis Type="Italic">Cerasiferum</Emphasis>)

The present study aimed to evaluate for the first time the phenols, flavonoids, carotenoids, capsaicin and dihydrocapsaicin content and the antioxidant and hypoglycemic properties of Capsicum annuum var. acuminatum small and C. annuum var. cerasiferum air-dried fruits. The ethanol extract of C. annuum var. acuminatum small, characterized by the major content of total poliphenols, flavonoids, carotenoids and capsaicinoids, showed the highest radical scavenging activity (IC₅₀ of 152.9 μg/ml). On the contrary, C. annuum var. cerasiferum showed a significant antioxidant activity evaluated by the β-carotene bleaching test (IC₅₀ of 3.1 μg/ml). The lipophilic fraction of both C. annuum var. acuminatum and C. annuum var. cerasiferum exhibited an interesting and selective inhibitory activity against α-amylase (IC₅₀ of 6.9 and 20.1 μg/ml, respectively).     

Comparative Study of Lipophilic and Hydrophilic Antioxidants from <Emphasis Type="Italic">In vivo</Emphasis> and <Emphasis Type="Italic">In vitro</Emphasis> Grown <Emphasis Type="Italic">Coriandrum sativum</Emphasis>

Coriander is commonly used for medicinal purposes, food applications, cosmetics and perfumes. Herein, the production of antioxidants in vegetative parts (leaves and stems) of in vivo and in vitro grown samples was compared. In vitro samples were clone A- with notorious purple pigmentation in stems and leaves and clone B- green. Seeds were also studied as they are used to obtain in vivo and in vitro vegetative parts. Lipophilic (tocopherols, carotenoids and chlorophylls) and hydrophilic (sugars, ascorbic acid, phenolics, flavonols and anthocyanins) compounds were quantified. The antioxidant activity was evaluated by radical scavenging activity, reducing power and lipid peroxidation inhibition. The in vivo sample showed the highest antioxidant activity mainly due to its highest levels of hydrophilic compounds. Otherwise, in vitro samples, mainly clone A, gave the highest concentration in lipophilic compounds but a different profile when compared to the in vivo sample. Clones A and B revealed a lack of β-carotene, β- and δ-tocopherols, a decrease in α-tocopherol, and an increase in γ-tocopherol and clorophylls in comparison to the in vivo sample. In vitro culture might be useful to explore the plants potentialities for industrial applications, controlling environmental conditions to produce higher amounts of some bioactive products.     

Rooting Characteristics of <Emphasis Type="Italic">Solanum chacoense</Emphasis> and <Emphasis Type="Italic">Solanum tuberosum in Vitro</Emphasis>

The precise level of environmental control in vitro may aid in identifying genetically superior plant germplasm for rooting characteristics (RC) linked to increased foraging for plant nitrogen (N). The objectives of this research were to determine the phenotypic variation in root morphological responses of 49 Solanum chacoense (chc), 30 Solanum tuberosum Group Phureja – Solanum tuberosum Group Stenotomum (phu-stn), and three Solanum tuberosum (tbr) genotypes to 1.0 and 0.5 N rate in vitro for 28 d, and identify genotypes with superior RC. The 0.5 N significantly increased density of root length, surface area, and tips. All RC were significantly greater in chc than in either phu-stn or tbr. Based upon clustering on root length, surface area, and volume, the cluster with the greatest rooting values consisted of eight chc genotypes that may be utilized to initiate a breeding program to improve RC in potato.     

Cupuassu (<Emphasis Type="Italic">Theobroma grandiflorum</Emphasis>) Peel as Potential Source of Dietary Fiber and Phytochemicals in Whole-Bread Preparations

Cupuassu (Theobroma grandiflorum) is a fruit tree native to the Brazilian Amazon. Cupuassu beans are extensively used in the Brazilian food industry. Fat from cupuassu beans, which are a rich source of triacylglycerols and fatty acids, is used extensively in the production of candies and confectionery in the northern and northeastern regions of Brazil. The potential use of the agro-industrial by-products of cupuassu has only slightly been addressed by the scientific community. Often, such by-products are sources of bioactive compounds with functional properties. Thus, the aims of this study were to characterize the use of cupuassu peel flour (CPF) and to examine the potential of CPF as a partial replacement in the preparation of breads through various means: chemical analyses, determination of protein digestibility, tannins, phytic acid and phenolic contents, pH, color, volume, and acceptance tests. The results show that CPF is a potential source of dietary fiber (79.81%), mainly insoluble fiber (78.29%), and breads made with added CPF present high dietary fiber content (5.40 and 6.15 g/100 g for inclusions with 6 and 9% CPF, respectively) and phytochemical values. The use of this by-product did not produce substantial changes in the physical, chemical or rheological characteristics of breads. Therefore, breads enhanced with CPF may be a convenient functional food, offering a good source of dietary fiber and phytochemicals. Breads prepared with 6% added CPF presented an acceptable overall quality to consumers.     

Antioxidant and Antimutagenic Activities of <Emphasis Type="Italic">Randia echinocarpa</Emphasis> Fruit

We report for the first time the antioxidant and antimutagenic activities of fractions from Randia echinocarpa fruit, which is a Rubiaceae plant native to Sinaloa, Mexico. This fruit has been traditionally used in the prevention or treatment of cancer, among other diseases. The pulp of the fruit was sequentially extracted with solvents of different polarity (i.e. hexane, chloroform, methanol and water). A high extraction yield was obtained with methanol (72.17% d.w.). The aqueous extract showed the highest content of phenolics (2.27 mg/g as ferulic acid equivalents) and the highest antioxidant activity based on the β-carotene bleaching method (486.15). The commercial antioxidant BHT was used as control (835.05). Antimutagenic activity of the aqueous extract (0–500 μg/tube) was evaluated using the Salmonella microsuspension assay (YG1024 strain) and 1-NP as the mutagen (50 and 100 ng/tube). The aqueous extract was neither toxic nor mutagenic and the percentage of inhibition on 1-NP mutagenicity was 32 and 53% at doses of 50 and 100 ng/tube, respectively. The results of the double incubation assay suggest that the extract inhibited the mutagenicity of 1-NP by a combination of desmutagenic and bioantimutagenic effects.     

Antioxidant and Antiproliferative Activity of <Emphasis Type="Italic">Diospyros lotus</Emphasis> L. Extract and Isolated Compounds

The object of the study was to determine the chemical composition of Diospyros lotus L. extract and their antioxidant and antiproliferative properties. Eight compounds were isolated from D. lotus and identified as gallic acid, methylgallate, ellagic acid, kaempferol, quercetin, myricetin, myricetin 3-O-β-glucuronide, and myricetin-3-O-α-rhamnoside. D. lotus extract tested in different in vitro systems (DPPH, ABTS, FRAP, and Fe²⁺ chelating activity assay) showed significant antioxidant activity. The potential antiproliferative properties of D. lotus extract and isolated compounds against nine human cancer cell lines such as COR-L23, CaCo-2, C32, ACHN, A375, A549, Huh-7D12, MCF-7, and LNCaP were investigated in vitro by SRB assay. D. lotus extract demonstrated the highest inhibitory activity against COR-L23 with an IC₅₀ value of 12.2 μg/ml. Among identified hydrolysable tannins, ellagic acid evidenced strong antiproliferative activity against both C32 and A375 cells with IC₅₀ values of 0.8 and 4.1 μg/ml, respectively. Interesting results were observed, also, with gallic acid that showed the highest cytotoxic activity against CaCo-2 (IC₅₀ 2.6 μg/ml). Overall, the results of this study suggest that D. lotus displays a good antioxidant activity and has antiproliferative effects. Both activities are related to identified phenolic compounds.     

Divergent evolution of rice blast resistance <Emphasis Type="Italic">Pi54</Emphasis> locus in the genus <Emphasis Type="Italic">Oryza</Emphasis>

Background

The rice blast resistance gene Pi54 was cloned from Oryza sativa ssp. indica cv. Tetep, which conferred broad-spectrum resistance against Magnaporthe oryzae. Pi54 allelic variants have been identified in not only domesticates but also wild rice species, but the majority of japonica and some indica cultivars lost the function.

Results

We here found that Pi54 (Os11g0639100) and its homolog Os11g0640600 (named as #11) were closely located on a 25 kbp region in japonica cv. Sasanishiki compared to a 99 kbp region in japonica cv. Nipponbare. Sasanishiki lost at least six genes containing one other R-gene cluster (Os11g0639600, Os11g0640000, and Os11g0640300). Eight AA-genome species including five wild rice species were classified into either Nipponbare or Sasanishiki type. The BB-genome wild rice species O. punctata was Sasanishiki type. The FF-genome wild rice species O. brachyantha (the basal lineage of Oryza) was neither, because Pi54 was absent and the orientation of the R-gene cluster was reversed in comparison with Nipponbare-type species. The phylogenetic analysis showed that #11gene of O. brachyantha was on the root of both Pi54 and #11 alleles. All Nipponbare-type Pi54 alleles were specifically disrupted by 143 and 37/44?bp insertions compared to Tetep and Sasanishiki type. In addition, Pi54 of japonica cv. Sasanishiki lost nucleotide-binding site and leucine-rich repeat (NBS–LRR) domains owing to additional mutations.

Conclusions

These results suggest that Pi54 might be derived from a tandem duplication of the ancestor #11 gene in progenitor FF-genome species. Two divergent structures of Pi54 locus caused by a mobile unit containing the nearby R-gene cluster could be developed before domestication. This study provides a potential genetic resource of rice breeding for blast resistance in modern cultivars sustainability.

     

Antioxidant in Mango (<Emphasis Type="Italic">Mangifera indica</Emphasis> L.) Pulp

This work was carried out to investigate the pulp composition of four mango cultivars (Haden, Tommy Atkins and Ubá) at the ripening stage in relation to three components with antioxidant potential (total phenolics, carotenoids and ascorbic acid). Total phenolic compound content was estimated by the Folin-Ciocalteu reagent and total carotenoid content by spectrophotometry at 450 nm. The contents of β-carotene and total vitamin C (ascorbic acid and dehydroascorbic acid) were quantified by high performance liquid chromatography. Differences were found among the four mango cultivars in all the components analyzed. The content of phenolic compounds ranged from 48.40 (Haden) to 208.70 mg/100 g (Ubá); total carotenoid from 1.91 (Haden) to 2.63 mg/100 g (Palmer); β-carotene from 661.27 (Palmer) to 2,220 μg/100 g (Ubá) and total ascorbic acid ranged from 9.79 (Tommy Atkins) to 77.71 mg/100 g (Ubá). These results corroborated previous information that mangoes are a good source of antioxidants in human diet.     

Potentials of Gum From <Emphasis Type="Italic">Detarium microcarpum</Emphasis> (DM) and <Emphasis Type="Italic">Mucuna flagellipes</Emphasis> (MF) Seeds as Raw Beef Burger Stabilizers

Raw beef burgers containing graded levels (0.25, 0.5, 0.75, and 1.0%) of polysaccharide gums extracted from Detarium microcarpum (DM) and Mucuna flagellipes (MF) were produced. Unstabilized beef burgers and beef burgers containing gum tragacanth (TR) were also produced simultaneously to serve as control. The raw beef burgers were evaluated for selected physicochemical and sensory properties so as to assess the stabilization potentials of DM and MF polysaccharide gums relative to the conventional TR. Beef burgers containing the polysaccharide gums had significantly (P<0.05) lower shrinkage, higher water holding capacity (WHC), and better stability under ambient conditions (27±1°C and RH 90.6%) then the unstabilized burgers. Among the stabilized burgers, DM gum burgers had lower (P0.05) shrinkage and higher WHC than MF and TR gums burgers. All the gum-stabilized beef burgers required 9–26% higher compression force indicating a less friable product than the unstabilized burgers. At 0.25 and 0.5% levels of gum incorporation DM gum burgers had comparable compression force with TR gum burgers and this was lower (P0.05) than the compression force of MF gum burgers. Sensory panel results indicated a lower score for finger feel in gum-stablized burgers than the unstabilized burgers. DM and TR gum burgers had comparable scores with TR gum burgers in all the sensory attributes tested. Overall acceptability score showed that all the gum-stabilized beef burgers were acceptable. It was evident from the study that polysaccharide gums from DM and MF seeds can serve as effective stabilizers in beef burgers without adverse effect on the quality of the product.     

Effect of Anthocyanin-Rich Extract from Black Rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L. <Emphasis Type="Italic">indica</Emphasis>) on Hyperlipidemia and Insulin Resistance in Fructose-Fed Rats

This study was designed to evaluate the effect of an anthocyanin-rich extract from black rice on hyperlipidemia and insulin resistance in fructose-fed rats. Rats fed fructose diet for 4 weeks exhibited significantly higher plasma insulin levels and lower insulin sensitivity than the control rats fed AIN-93G diet. Dietary supplementation with the anthocyanin-rich extract (5 g/kg of high-fructose diet) prevented the development of fructose-induced insulin resistance. After fructose-induced insulin resistance had been established, 4-week treatment with the anthocyanin-rich extract (5 g/kg of high-fructose diet) or pioglitazone (270 mg/kg of high-fructose diet) ameliorated the glucose intolerance and hyperlipidemia, but the extract failed to reverse the fructose-induced hyperinsulinemia as pioglitazone did. In addition, rats supplemented by the extract exhibited lower oxidative stress than the fructose-fed controls, as indicated by the lower concentrations of plasma thiobarbituric acid reactive substances and blood oxidized glutathione. Overall, these results suggest that the anthocyanin-rich extract from black rice improves certain metabolic abnormalities associated with diets high in fructose.