<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.     

Volatile Compounds and Antioxidative Activity of <Emphasis Type="Italic">Porophyllum tagetoides</Emphasis> Extracts

Porophyllum tagetoides is an annual warm-weather herb that has an intense typical smell. Its leaves are commonly used in soup preparation and traditional medicine for treatment of inflammatory diseases. Its volatile compounds and antioxidant properties were evaluated in crude, aqueous and ethanol leaf extract and an oil emulsion using different antioxidant assays in vitro, such as: DPPH radical scavenging activity, redox potential, polyphenol content, reducing power and optical density. A high antioxidative activity was found when comparing leaves with stems. The crude extract from leaves showed a very high reducing power (2.88 ± 0.20 O.D.) and DPPH radical-scavenging activity (54.63 ± 4.80%), in concordance with a major concentration of vitamin C (23.97 ± 0.36 mg/100 g). Instead, the highest polyphenol content (264.54 ± 2.17 mg GAE/g of sample) and redox potential (561.23 ± 0.15 mV) were found by the ethanol and aqueous extract, respectively. Aldehydes and terpenes such as nonanal, decanal, trans-pineno, β-myrcene and D-limonene were the major volatiles found. This study suggests that Porophyllum tagetoides extracts could be used as antioxidants.     

<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.     

Kinetics of the Inhibition of Renin and Angiotensin I Converting Enzyme by Polar and Non-polar Polyphenolic Extracts of <Emphasis Type="Italic">Vernonia Amygdalina</Emphasis> and <Emphasis Type="Italic">Gongronema Latifolium</Emphasis> Leaves

The aim of this study was to determine the in vitro modulation of the renin-angiotensin system by polyphenolic extracts and fractions of two green leafy vegetables, Vernonia amygdalina (VA) and Gongronema latifolium (GL), that are used for food and medicinal purposes. An 80% acetone extract of each leaf was fractionated on silicic acid-packed column to give two main fractions: acetone eluate (flow-through) and ethanol eluate (column-bound), that consist mostly of chlorophyllic and non-chlorophyllic fractions, respectively. Column fractionation resulted in polyphenolic fractions that displayed higher potency against angiotensin converting enzyme (ACE) and renin than the crude acetone extracts; generally, the chlorophyllic fraction was more active than the non-chlorophyllic fraction. ACE-inhibitory activity was significantly higher (p < 0.05) for the chlorophyllic fraction of VA than GL, with IC₅₀ values of 0.207 and 0.413 mg/ml, respectively. Similarly, the chlorophyllic fraction of VA had significantly higher (p < 0.05) renin inhibition than GL, with IC₅₀ values of 0.172 and 0.513 mg/ml, respectively. Kinetics studies showed that the chlorophyllic fractions of VA and GL exhibited mostly mixed-type ACE and renin inhibitions. We concluded that the hydrophobic nature of the chlorophyllic fraction may have contributed to the increased interaction with enzyme protein and inhibition of activities of ACE and renin.     

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.     

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.     

<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.     

Hypolipidemic Effect of Avocado (<Emphasis Type="Italic">Persea americana</Emphasis> Mill<Emphasis Type="Italic">)</Emphasis> Seed in a Hypercholesterolemic Mouse Model

Avocado seed contains elevated levels of phenolic compounds and exhibits antioxidant properties. We investigated the effect of Avocado Seed Flour (ASF) on the lipid levels in mice on a hyperlipidemic diet. The concentration of phenols was determined by high-performance liquid chromatography, antioxidant activity was evaluated using the Trolox equivalent antioxidant capacity method, and dietary fiber was measured using the Association of Official Analytical Chemists (AOAC) method. The LD₅₀ of ASF was determined using Lorke’s method and hypolipidemic activity was evaluated in a hypercholesterolemic model in mice. Protocatechuic acid was the main phenolic compound found in ASF, followed by kaempferide and vanillic acid. The total phenolic content in the methanolic extract of ASF was 292.00 ± 9.81 mg gallic acid equivalents/g seed dry weight and the antioxidant activity resulted in 173.3 μmol Trolox equivalents/g DW. In addition, a high content of dietary fiber was found (34.8%). The oral LD₅₀ for ASF was 1767 mg/kg body weight, and treatment with ASF significantly reduced the levels of total cholesterol, LDL-C, and prediction of the atherogenic index. Therefore, the antioxidant activity of phenolic compounds and dietary fiber in ASF may be responsible for the hypocholesterolemic activity of ASF in a hyperlipidemic model of mice.     

<Emphasis Type="Italic">In vitro</Emphasis> Starch Digestibility and Predicted Glycemic Index of Microwaved and Conventionally Baked Pound Cake

The present study compares the effect of baking process (microwave vs conventional oven) on starch bioavailability in fresh pound cake crumbs and in crumbs from pound cake stored for 8 days. Proximal chemical analysis, resistant starch (RS), retrograded starch (RS3) and starch hydrolysis index (HI) were evaluated. The empirical formula suggested by Granfeldt was used to determine the predicted glycemic index (pGI). Pound cake, one of Mexico’s major bread products, was selected for analysis because the quality defects often associated with microwave baking might be reduced with the use of high-fat, high-moisture, batted dough. Differences in product moisture, RS and RS3 were observed in fresh microwave-baked and conventionally baked pound cake. RS3 increased significantly in conventionally baked products stored for 8 days at room temperature, whereas no significantly changes in RS3 were observed in the microwaved product. HI values for freshly baked and stored microwaved product were 59 and 62%, respectively (P > 0.05), whereas the HI value for the conventionally baked product decreased significantly after 8 days of storage. A pound cake with the desired HI and GI characteristics might be obtained by adjusting the microwave baking process.     

Evaluation of the Biochemical Components and Chromatic Properties of the Juice of <Emphasis Type="Italic">Vaccinium macrocarpon</Emphasis> Aiton and <Emphasis Type="Italic">Vaccinium oxycoccos</Emphasis> L.

Benzoic acid, total anthocyanins, soluble solids, titratable acidity, and colour properties in juice of the American cranberry Vaccinium macrocarpon and the European cranberry Vaccinium oxycoccos were investigated. Berry juices of V. macrocarpon cultivars were distinguished by their higher total anthocyanin and benzoic acid amounts. These cultivars accumulated on average 43.11 mg/l of benzoic acid and 92.45 mg/l of total anthocyanins. The levels of benzoic acid and total anthocyanins in V. oxycoccos cultivars were 17.52 mg/l and 42.54 mg/l, respectively. The V. macrocarpon cultivars ‘Franklin’, ‘Le Munyon’, ‘Searles’, and ‘Early Richard’ were selected as the best according to the enhanced total anthocyanins and benzoic acid amounts. The separation of anthocyanins by HPLC-UV-VIS revealed the presence of six anthocyanins, with peonidin-3-galactoside being the most prevalent. Galactoside together with glucoside conjugates comprised the largest percentage of total anthocyanins in the juices of V. macrocarpon and V. oxycoccos cultivars.     

Superoxide Dismutase Activity in Mesocarp Tissue from Divergent <Emphasis Type="Italic">Cucumis melo</Emphasis> L. Genotypes

Muskmelons (Cucumis melo L.) are well-known as excellent sources of several vitamins, minerals and non-enzymatic antioxidant phytochemicals such as vitamin C and pro-vitamin A. Less well-studied is their potential role as sources of enzymatic antioxidants such as superoxide dismutase (SOD), which have been associated with enhanced reactive oxygen species scavenging capacity in some muskmelon fruits. In this study, we investigated the variability in SOD activities among diverse advanced breeding lines and commercial muskmelon cultivars grown in two different soil types—clay or sandy loam. Specific and total SOD activities varied significantly among the genotypes (P ≤ 0.001), compared to soil type (P ≤ 0.055). Netted (cantaloupe) genotypes generally had the lowest SOD activities compared to the green- and orange-fleshed honey dew types. Casaba type fruit had average SOD activities that were ∼ 1.6-fold greater than those of honey dew types, and ∼ 9.0-fold greater than those of cantaloupe types. These data indicate there is useful genetic diversity among commercial melon varieties and in exotic genotypes that could be used to develop C. melo as a functional food with enhanced SOD content.     

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).     

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.     

Acute Effects of <Emphasis Type="Italic">Ginkgo Biloba</Emphasis> Extract on Vascular Function and Blood Pressure

We investigated whether a single dose of standardized Ginkgo biloba extract (GBE) can improve vascular function. A randomised controlled crossover trial was conducted on 14 young healthy men, who received GBE or placebo. The digital volume pulse was monitored to measure reflection index (DVP-RI) and stiffness index (DVP-SI) and peripheral augmentation index (pAIx) was assessed using radial pulse wave analysis at baseline and 2, 4 and 6 h after treatment. DVP-SI was slightly higher 2 h following GBE compared to placebo (P < 0.05); other outcome variables were unaffected by treatment.     

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).     

<Emphasis Type="Italic">Carica papaya</Emphasis> Lipase: A Naturally Immobilized Enzyme with Interesting Biochemical Properties

Triacylglycerol (TAG) lipases have been thoroughly characterized in mammals and microorganisms, whereas very little is known about plant TAG lipases. The lipolytic activity occurring in all the laticies is known to be associated with sedimentable particles, and all attempts to solubilize the lipolytic activity of Carica papaya latex have been unsuccessful so far. However, some of the biochemical properties of the lipase from Carica papaya latex (CPL) were determined from the insoluble fraction of the latex. The activity was optimum at a temperature of 37°C and a pH of 9.0, and the specific activities of CPL were found to be 2,000 ± 185 and 256 ± 8 U/g when tributyrin and olive oil were used as substrates, respectively. CPL was found to be active in the absence of any detergent, whereas many lipases require detergent to prevent the occurrence of interfacial denaturation. CPL was inactive in the presence of micellar concentrations of Triton X-100, sodium dodecyl sulfate (SDS) and tetradecyl trimethylammonium bromide (TTAB), and still showed high levels of activity in the presence of sodium taurodeoxycholate (NaTDC) and the zwitterionic Chaps detergent. The effects of various proteases on the lipolytic activity of CPL were studied, and CPL was found to be resistant to treatment with various enzymes, except in the presence of trypsin. All these properties suggest that CPL may be a good candidate for various biotechnological applications.     

Absorption,Distribution and Elimination of Carnosic Acid,A Natural Antioxidant from <Emphasis Type="Italic">Rosmarinus officinalis</Emphasis>, in Rats

The absorption, distribution and elimination of carnosic acid, the main antioxidant found in rosemary was studied, in vivo, in rats. Therefore, carnosic acid was administrated in a single dose, intravenously (20.5 ± 4.2 mg/kg) and orally (64.3 ± 5.8 mg/kg), to four and nine rats, respectively. Blood samples were collected at different time points, and plasma concentrations of carnosic acid were determined using LC-MS. Furthermore, total collection of urine and feces was done during 4 h and 24 h for the intravenous and oral administrations, respectively. After euthanizing the rats, intestinal content, liver and muscle tissue were sampled to determine carnosic acid concentrations. The bioavailability of carnosic acid, after 360 min, was 40.1%. Traces of carnosic acid were found in the rats intestinal content, liver and muscle tissue of abdomen and legs. The recovery of carnosic acid in the feces, 24 h after oral administration, was 15.6 ± 8.2%. Carnosic acid is absorbed into the bloodstream after oral administration in rats and is therefore bioavailable. It was found that carnosic acid in vivo is present in its free form and that its main elimination route is the fecal route.     

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.     

Nutrient Content of Carob Pod (<Emphasis Type="Italic">Ceratonia siliqua</Emphasis> L.) Flour Prepared Commercially and Domestically

Although the fruit of the carob tree (Ceratonia siliqua L. Fabaceae) is nutritious and widely available in Turkey, especially in West and South Anatolia, much remains to be learned about its nutrient composition. The main goal of our study was to determine if there are differences in the content of certain nutrients in commercially-prepared carob flour (CPCP) and domestic or home-prepared carob powder (HPCP). Sucrose was the main sugar in CPCP and HPCP. Total protein was 40% lower in CPCP than HPCP due mainly to decreases in the content of several essential amino acids. However, except for lysine in CPCP, HPCP and CPCP compared favourably to a WHO protein standard. There were large differences in terms of their content of the two essential fatty acids, linoleic and α-linolenic acid, and the linoleic acid/α-linolenic acid ratio was 3.6 for CPCP, and 6.1 for HPCP. Manganese and iron were 2.5-fold higher in HPCP than CPCP. This study demonstrates that carob flour prepared in either the household or industrially is a good source of many, but not all essential nutrients, and that commercial processing of carob fruit into flour seems to affect its content of several important nutrients.