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.     

Vitamin C Enrichment of Fruit Juice Based Ready-to-Serve Beverages Through Blending of Indian Gooseberry (<Emphasis Type="Italic">Emblica officinalis</Emphasis> Gaertn.) Juice

Physiconutritional qualities of fruits viz. apple, lime, pome- granate, Perlette grape, and Pusa Navrang grape were analyzed and compared with those of Indian gooseberry (Emblica officinalis Gaertn.). Indian gooseberry juice contained the highest vitamin C (478.56 mg/100 ml). Hence, when gooseberry juice was blended with other fruits juice for the preparation of ready-to-serve (RTS) beverages, it boosted their nutritional quality in terms of vitamin C content. On the basis of overall sensory quality and vitamin C content, RTS beverage prepared by blending gooseberry and Pusa Navrang grape juice in 20:80 ratio was found to be the best.     

Antioxidative and Anticancer Activity of Extracts of Cherry (<Emphasis Type="Italic">Prunus serrulata</Emphasis> var. <Emphasis Type="Italic">spontanea</Emphasis>) Blossoms

Organic solvent (methanol, ethanol, and acetone) extracts and water extracts of cherry (Prunus serrulata var. spontanea) blossoms were prepared, and antioxidant activities of the extracts were evaluated. Methanolic CBE (100 μg/ml) showed the highest total phenol content (104.30 μM), radical scavenging activity (34.2%), and reducing power (0.391). The effect of CBE on DNA damage induced by H₂O₂ in human leukocytes was evaluated by Comet assay. All CBE was a potent dose dependent inhibitor of DNA damage induced by 200 μM of H₂O₂, methanolic CBE showed the most strong inhibition activity. The methanolic CBE of 500 μg/ml showed 38.8% inhibition against growth of human colon cancer cell line HT-29. These results indicated that cherry blossoms could provide valuable bioactive materials.     

Analysis of Flavonoids from Leaves of Cultivated <Emphasis Type="Italic">Lycium barbarum</Emphasis> L.

Leaves of Lycium barbarum are widely used as medicine vegetables and functional tea in China. The main flavonoids present in the leaves were separated and identified by high performance liquid chromatography (HPLC), liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-(APCI) MS) and ultraviolet-visible spectra with shift additives. The predominant flavonoid was identified as rutin. Leaves are the rutin-rich parts (16.03–16.33 mg/g). In the wild and cultivated L. barbarum fruits, contents of rutin were determined very low (0.09–1.38 mg/g). The contents of total flavonoids (21.25 mg/g) of cultivated L. barbarum leaves were much higher than those in the wild L. barbarum leaves (17.86 mg/g), so cultivated barbarum leaves are a suitable source for medicine vegetables and functional tea.     

<Emphasis Type="Italic">Pulque</Emphasis>, An Alcoholic Drink from Rural Mexico,Contains Phytase. Its <Emphasis Type="Italic">in vitro</Emphasis> Effects on Corn Tortilla

Pulque is made by fermenting the agave sap or aguamiel of Agave atrovirens with a whole array of microorganisms present in the environment including several lactic acid bacteria and yeasts such as Saccharomyces cerevisiae. Ascorbic acid was determined in pulque and aguamiel, respectively. Phytase activity in lees, liquid and freeze-dried pulque was assayed by measuring the appearance of phosphate from phytate by a colorimetric method likewise phosphate from phytate present in fresh corn tortilla was measured after in vitro incubation with pulque. Iron, zinc, calcium, magnesium and selenium contents were measured in pulque and corn tortilla as well as in nixtamalized corn flour (NCF), the latter is used to make instant tortilla, since corn provides most of the energy as well as most of the phytate in the Mexican rural diet. Pulque showed phytase activity but much less ascorbic acid and iron than previously reported; additionally, phytase in pulque hydrolyzed most of phytate’s corn tortilla. Lees, which is mostly made of pulque’s microbiota, significantly accumulated iron and zinc but no selenium. NCF was fortified with iron by the manufacturers but poorly blended. There were significant differences on selenium content between tortillas samples, apparently some soils in central Mexico are selenium deficient. Moderate pulque intake appears to increase the bioavailability of iron and zinc bound by phytate in corn.     

Rutin and Total Quercetin Content in Amaranth (<Emphasis Type="Italic">Amaranthus</Emphasis> spp.)

The aim of the study was to confirm the presence of rutin, one of the most common quercetin glycosides, and other quercetin derivatives in plants of genus Amaranthus, to investigate the influence of the species and variety on rutin distribution in the plant and content changes during growing season. The rutin content was determined by micellar electrokinetic capillary chromatography in individual plant parts at the beginning of the growth, at the flowering stage and at the maturity stage of five Amaranthus species. The total quercetin content was determined by micellar electrokinetic capillary chromatography too. The rutin content in amaranth ranged from 0.08 (in seeds) to 24.5 g/kg dry matter (in leaves). Comparison of the determined total quercetin content and the calculated content of quercetin released from rutin did not prove important presence of quercetin or other quercetin derivatives than rutin. Only amaranth leaves sampled at the maturity stage probably contained quercetin or quercetin derivatives. Significant differences in the rutin content were established among species and as well varieties. Amaranthus hybrid and A. cruentus were the best sources of rutin.     

Antioxidant and Antifungal Activity of <Emphasis Type="Italic">Verbena officinalis</Emphasis> L. Leaves

The scavenging activity against DPPH (1,1-diphenil-2-picrylhydrazyl) radical and the antifungal effect against chloroform, ethyl acetate and 50% methanolic extracts of Verbena officinalis leaves were investigated. The activity of different fractions of 50% methanolic extract and some isolated compounds were also investigated. The results suggest that 50% methanolic extract and caffeoyl derivatives could potentially be considered as excellent and readily available sources of natural antifungal and antioxidant compounds.     

Phenolics,Sugars, Antimicrobial and Free-Radical-Scavenging Activities of <Emphasis Type="Italic">Melicoccus bijugatus</Emphasis> Jacq. Fruits from the Dominican Republic and Florida

Edible fruits of the native South American tree Melicoccus bijugatus Jacq. are consumed fresh or in traditional food, drink and medicinal preparations. Some therapeutic effects of these fruits may be due to phenolics and sugars. Aqueous acetone, methanol or ethanol tissue extracts of different cultivars or collections of M. bijugatus fruits from the Dominican Republic and Florida were analyzed for total phenolics and free radical scavenging activity by UV-vis spectroscopy, sugars by gas chromatography, and antimicrobial activity by the disc diffusion assay. Total phenolics and free radical scavenging activities ranked: seed coat > embryo > pulp extracts. Montgomery cultivar fruits had the highest total phenolics. For sugars: pulp > embryo and highest in Punta Cana fruit pulp. In all extracts: sucrose > glucose and fructose. Glucose:fructose ratios were 1:1 (pulp) and 0.2:1 (embryo). Pulp extracts had dose-response antibacterial activity and pulp and embryo extracts had antifungal activity against one yeast species. Phenolics and sugars were confirmed with thin-layer chromatography and nuclear magnetic resonance. Sugar-free pulp fractions containing phenolics had slightly more antimicrobial activity than H₂O-soluble pulp fractions with sugars. Results indicate M. bijugatus fruits contain phenolics, sugars and other H₂O-soluble compounds consistent with therapeutic uses. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Quantification of Zeaxanthin Dipalmitate and Total Carotenoids in <Emphasis Type="Italic">Lycium</Emphasis> Fruits (Fructus Lycii)

An HPLC-DAD method has been developed to quantitatively analyze for the content of zeaxanthin dipalmitate, a major carotenoid in Fructus Lycii, in different species of the genus Lycium. Determination was performed using an Alltima C18 column with the mobile phase consisting of acetonitrile and dichloromethane (42:58). The total contents of carotenoids in these samples were also determined by using UV spectrophotometric assay. Total carotenoid concentrations of different Fructus Lycii are within the range of 0.03–0.5%. Zeaxanthin dipalmitate is a predominant carotenoid, comprising 31–56% of the total carotenoids in Fructus Lycii. This study is the first systematic quantification of the carotenoids in the fruits of different Lycium species. The results demonstrated that these methods are reliable and facile techniques for rapid analysis of carotenoids for crude drug and plant-derived food supplements.     

Antioxidant,Antimicrobial and Genotoxicity Screening of Hydro-alcoholic Extracts of Five Serbian <Emphasis Type="Italic">Equisetum</Emphasis> Species

The hydro-alcoholic extracts of five Equisetum species, E. arvense L., E. sylvaticum L., E. fluviatile L., E. palustre L. and E. telmateia Ehrh., growing-wild in Serbia were evaluated for their genotoxicity, antimicrobial activity, antioxidant capacity and the results related to the total phenol content and HPLC flavonoid profiles. The total phenol content was 92–349 μmol expressed as equivalents of chlorogenic acid per g of dried plant material. Main identified compounds were kaempferol-, quercetin- glycosides and caffeic acid derivatives. E. telmateia extract showed the greatest antioxidant capacity. Almost all tested microorganisms demonstrated some degree of sensitivity to the examined extracts. All tested extracts at 62.5 μg/ml showed higher incidence of micronucleus formation than in the control sample. The obtained data allowed mutual comparison of examined species and their assessment as possible sources of antioxidants, antimicrobials and/or genotoxic substances.     

Endosperm-Specific Expression of Serotonin <Emphasis Type="Italic">N</Emphasis>-Hydroxycinnamoyltransferase in Rice

Serotonin N-hydroxycinnamoyltransferase (SHT) is a key enzyme in the synthesis of feruloylserotonin (FS) and 4-coumaroylserotonin (CS). These serotonin derivatives show strong antioxidant activity, making them valuable for both nutritional and pharmacological use in humans. Ectopic expression of SHT under the control of the endosperm specific-glutelin and prolamin promoters from rice was produced via Agrobacterium-mediated transformation. SHT expression was confirmed by Southern blot analysis, followed by Northern blotting and SHT enzyme activity analyses using total RNA and protein, respectively, extracted from transgenic seeds. The glutelin A3 (GluA3) promoter produced low SHT mRNA expression in rice seeds, whereas the prolamin promoter expressed high levels of SHT mRNA. In spite of the ectopic expression of SHT in rice seeds, both transgenic genotypes accumulated levels of serotonin derivatives similar to those found in wild-type rice. Furthermore, our data suggest that serotonin, rather than phenylpropanid-CoAs, is the rate-limiting substrate in the biosynthesis of serotonin derivatives in SHT-overexpressing transgenic rice seeds.     

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.     

Physicochemical,Nutritional and Antibacterial Characteristics of the Fruit of <Emphasis Type="Italic">Bromelia pinguin</Emphasis> L.

Bromelia pinguin L. is a plant native to Sinaloa, Mexico, where its fruit is used as food or as a phytotherapeutic agent. The fruits of B. pinguin were characterized and they could be considered as a functional food. These fruits show an average weight of 13.7 g and a yellow color of high luminosity (b* = 43.2, L* = 74.5). The values for acidity (4.6%, as citric acid) and pH (3.7) of B. pinguin fruit are similar to those of citrics. The edible portion is characterized by a high content of vitamin C (126 mg/100 g), ash (10.6 g/100 g d.w.), crude fiber (3.4 g/100 g d.w.), calcium (1290 mg/100 g d.w.), magnesium (500 mg/100 g d.w.), manganese (2.95 mg/100 g d.w.) and it is a good source of zinc (2.8 mg/100 g d.w.). Polar fractions extracted from the pulp fruit showed activity against several genera of human pathogenic-bacteria (Staphylococcus, Streptococcus, Enterococcus, Salmonella, Shigella, Escherichia and Pseudomonas).     

Applied Biotechnology to Combat Late Blight in Potato Caused by <Emphasis Type="Italic">Phytophthora Infestans</Emphasis>

Potato is an important crop, grown worldwide. It suffers from many pests and diseases among which late blight, caused by the oomycete Phytophthora infestans, is the worst. The disease is still causing major damage in many potato production areas and control is only possible by applying fungicides frequently. The knowledge on the molecular biology and genetics of the interaction between the plant and the oomycete is developing rapidly. These are relevant fields of study, currently dominated by the discovery of many resistance genes and numerous effector proteins and the analysis of their specific mode of action. These studies may yield essential information needed for the development of durable resistance. The long-term and worldwide effort to breed for resistance so far has had little effect. A novel breeding approach may change this. It is based on cisgenic modification (CM) consisting of marker-free pyramiding of several resistance genes and their spatial and temporal deployment yielding dynamic varieties that contain potato genes only. It is envisioned that this CM approach with potato’s own genes will not only prove societally acceptable but may also result in simplifications in the legislation on use of the CM approach. Various parties in the potato research arena intend to cooperate in this novel approach in a number of developing countries where potato substantially contributes to food security. The use of resources such as land, water and energy improves when the effect of late blight is markedly reduced.     

Composition and Functional Properties of <Emphasis Type="Italic">Lupinus campestris</Emphasis> Protein Isolates

Protein isolates from L. campestris and soybean seeds were prepared using isoelectric precipitation (PI) and micellization (MI) procedures. The amount of protein recovered was considerably higher with the isoelectric precipitation than with the micellization procedure (60% and 30%, respectively). Protein contents were higher than 90% in protein isolates. Antinutritional factors content (alkaloids, lectins, and tannins) were reduced to innocuous levels after protein isolate preparation. Minimum protein solubility for the precipitated lupin protein isolate (LPI) was at pH 4.0, and between pH 4 and 6 for the micellized lupin protein isolate (LMI), increasing at both extremes of the pH scale. Water absorption for the LMI was 1.3 ml/g of protein and its oil absorption 2.2 ml/g of protein. The LPI had 1.7 ml/g of protein in both water and oil absorption. Foaming capacity and stability was pH-dependent. Foaming capacity was higher at pH 2 and lower near the protein isoelectric points. Minimum protein concentration for gelation in LMI was 8% w/v at pH 4, while for LPI was 6% at pH 4 and 6. Amino acid composition in L. campestris flour and protein isolates was high in lysine and low in methionine. Most of the essential amino acids in lupin protein isolates were at acceptable levels compared to a reference pattern for infants and adults. The electrophoretic pattern of both protein isolates showed three bands with different mobilities, suggesting that the protein fractions belong to alpha-conglutin (11S-like protein), beta-conglutin (7S-like protein) and gamma-conglutin. It is proven that some of the functional properties of L. campestris protein isolates are similar to those soybean protein isolates recovered under equal conditions.     

Variation in the Carotenoid Composition of the Lycopene-Rich Brazilian Fruit <Emphasis Type="Italic">Eugenia uniflora</Emphasis> L.

The indigenous pitanga (Eugenia uniflora L.) is now marketed and commercially processed in Brazil. In the present work, the carotenoids of the pitanga fruit from two states and at two stages of ripening, as well as of processed pitanga products (frozen pulp and juice, the brands being designated as A, B and C) commercialized in Campinas, São Paulo, were determined by HPLC. As compared to ripe pitanga from Medianeira, Paraná, those from Campinas had significantly higher (all-E)-lycopene (14.0 vs. 71.1 μg/g), (13Z)-lycopene (1.1 vs. 5.0 μg/g) and (all-E)-γ-carotene (1.6 vs. 3.8 μg/g) levels. Significant increases in most of the carotenoids occurred from the partially ripe to the ripe fruits, with (all-E)-lycopene doubling its concentration in fruits from both states. Pitanga was found to be one of the richest fruit sources of carotenoids, particularly lycopene, but the processed products had much lower lycopene content. The mean (all-E)-lycopene concentration was 16.6 μg/g for frozen pulp brand A, 23.0 μg/g for bottled juice brand B and 25.6 μg/g for bottled juice brand C. Optimization of processing is therefore needed to guarantee better retention of this important carotenoid.     

The Consumption of <Emphasis Type="Italic">Hibiscus sabdariffa</Emphasis> Dried Calyx Ethanolic Extract Reduced Lipid Profile in Rats

The scientific basis for the statement that plants and their active constituents play an important role in the prevention of chronic and degenerative diseases is continously advancing. The object of the present study was to evaluate the effect of Hibiscus sabdariffa L. dried calyx ethanolic extract on the serum lipid profile of Sprague-Dawley rats. The rats were fed during 4 weeks with either a basal diet, containing high cholesterol (1%), cholic acid (0.25%), lard oil (10%), or a supplemental diet with H. sabdariffa extract at 5%, 10%, and 15% levels (SD₅, SD₁₀, SD₁₅). Weight gain and faeces dry weight were both very significantly less (p ≤ 0.01) in SD₁₀ and SD₁₅ groups as compared to the control group, as was food efficiency in SD₁₅. In the case of SD₁₀, food efficiency was only significantly lower (p ≤ 0.05). Faeces dry weight in SD₅ however, was the only one found to be significantly higher (p ≤ 0.05). Triacylglycerols and LDL levels were both significantly less (p ≤ 0.05) in all groups (SD₅, SD₁₀, and SD₁₅) as compared to the control. For total lipids, SD₁₀ and SD₁₅ showed significantly lower levels (p ≤ 0.05), whereas very significant differences (p ≤ 0.01) were observed in the case of SD₅. All groups had lower cholesterol levels compared to controls; however, only the SD₅ group was statistically significant (p ≤ 0.05). No significant results were found in any group in the cases of either phospholipid or HDL levels. A hypothesis of hibiscus acid racemization, (+)-HCA to (−)-HCA, mediated by intestinal flora enzymes possibly explains the significant triacylglycerol decrease in all experimental groups. VLDL, the precursor of LDL, is composed predominantly of triacylglycerols, suggesting that the significant decrease in LDL is related to observed triacylglycerol synthesis inhibition. Five percent H. sabdariffa extract addition showed the best results in the reduction of serum lipids under study conditions.     

Antiproliferation and Apoptosis on RKO Colon Cancer by <Emphasis Type="Italic">Millingtonia hortensis</Emphasis>

Millingtonia hortensis is a medicinal plant widely used in many Asian countries. An aqueous crude extract of this plant has been shown the apoptosis induction on RKO colon cancer cells. However, its mechanism remains unknown. To learn more about this plant extract, we partially purified the crude extract using Sephadex LH-20 and three aqueous fractions were collected. Each fraction was investigated for cytotoxicity using MTT assay. Fraction 1 showed antiproliferative effect on RKO cells with dose-dependent manner, while fraction 2 and 3 had no effect. Induction of apoptosis was determined using flow cytometry and DNA fragmentation method. Apoptotic cell numbers and the appearance of fragmented DNA increased with dose-dependent manner after treatment with fraction 1 for 48 h. We further investigated the expression of apoptotic protein by western blot analysis. Fraction 1 decreased the expression of anti-apoptotic protein, Bcl-x_L and p-Bad, while pro-apoptotic protein Bad, was not changed. Fraction 1 also decreased the expression of p-Akt and slightly increased the level of total Akt. These results indicated that fraction 1 is able to inhibit cell proliferation and induce apoptosis on RKO cells by decreasing the expression of Bcl-x_L, p-Bad and p-Akt which are involving in survival of cancer cells.     

<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.     

Antioxidant Activity and Chemical Characterization of Essential Oil of <Emphasis Type="Italic">Bunium persicum</Emphasis>

The search for natural antioxidants, especially of plant origin, has notably increased in recent years. Bunium persicum Boiss. is an economically important medicinal plant growing wild in the dry temperature regions in Iran. In this study, chemical constituents of the essential oil of the seed from Bunium persicum Boiss. have been studied by GC/MS technique. The major components were caryophyllene (27.81%), gamma-terpinene (15.19%), cuminyl acetate (14.67%). Individual antioxidant assays such as, DPPH* scavenging activity and beta-carotene bleaching have been carried out. In DPPH* system, the EC(50) value of essential oil was determined as 0.88 mg/mL. In beta-carotene bleaching antioxidant activity of essential oil (0.45%) was almost equal to BHT at 0.01%. In addition, the antioxidant activity of the essential oil was evaluated in crude soybean oil by monitoring peroxide and thiobarbituric acid values of the oil substrate. The results showed that the Bunium persicum essential oil (BPEO) was able to reduce the oxidation rate of the soybean oil in the accelerated condition at 60 degrees C (oven test). The essential oil at 0.06% showed the same effect of BHA at 0.02%. Hence, BPEO could be used as an additive in food after screening.