Lactic Acid Fermentation of Cactus Cladodes (Opuntia ficus-indica L.) Generates Flavonoid Derivatives with Antioxidant and Anti-Inflammatory Properties

Cactus pear (Opuntia ficus-indica L.) is widely distributed in the arid and semi-arid regions throughout the world. In the last decades, the interest towards vegetative crop increased, and cladodes are exploited for nutraceutical and health-promoting properties. This study aimed at investigating the capacity of selected lactic acid bacteria to increase the antioxidant and anti-inflammatory properties of cactus cladodes pulp, with the perspective of producing a functional ingredient, dietary supplement or pharmaceutical preparation. Preliminarily, the antioxidant activity was determined through in vitro assays. Further, it was confirmed through ex vivo analysis on intestinal Caco-2/TC7 cells, and the profile of flavonoids was characterized. Cactus cladode pulp was fermented with lactic acid bacteria, which were previously selected from plant materials. Chemically acidified suspension, without bacterial inoculum and incubated under the same conditions, was used as the control. Lactobacillus plantarum CIL6, POM1 and 1MR20, Lactobacillus brevis POM2 and POM4, Lactobacillus rossiae 2LC8 and Pediococcus pentosaceus CILSWE5 were the best growing strains. Fermentation of cladode pulp with L. brevis POM2 and POM4 allowed the highest concentration of γ-amino butyric acid. Lactic acid fermentation had preservative effects (P<0.05) on the levels of vitamin C and carotenoids. Two flavonoid derivatives (kaemferol and isorhamnetin) were identified in the ethyl acetate extracts, which were considered to be the major compounds responsible for the increased radical scavenging activity. After inducing oxidative stress by IL-1β, the increased antioxidant activity (P<0.05) of fermented cladode pulp was confirmed using Caco-2/TC7 cells. Fermented cladode pulp had also immune-modulatory effects towards Caco-2 cells. Compared to the control, fermented cladode pulp exhibited a significantly (P<0.05) higher inhibition of IL-8, TNFα and prostaglandins PGE2 synthesis. The highest functional effect was found using ethyl acetate extracts. In conclusion, fermentation, especially with L. plantarum strains and L. brevis POM4, enhanced the antioxidant and immune-modulation features of cladode pulp.     

Vegetative Parthenocarpy in the Cactus Pear Opuntia ficus-indica (L.) Mill.

The dependence of fruit development on fertilization was studiedin two clones of Opuntia ficus-indica, Ofer and BSI. Fruitsof the clone Ofer bear fully developed seeds, whereas fruitsof the clone BSI contain only degenerated seeds and it was suspectedthat BSI is parthenocarpic. The two clones differed in theirpattern of fruit development. The increase in fruit fresh weightin Ofer was a result of both peel and pulp growth, whereas inBSI fruit growth was mainly due to pulp growth and the peelhad reached its final weight almost at anthesis. Pulp growthin BSI commenced earlier and was faster than in Ofer, but thefinal pulp weight in BSI was only 64% of that in Ofer. Seedgrowth in BSI was limited to the development of semilignifiedand lignified seed coats, whereas in Ofer 43·5% of theseeds were completely developed, the remainder being similarto those found in BSI. Fruits bearing well-developed seeds werealso found in a 'mutant' of BSI that, like Ofer, contained smallerovules at anthesis than flower of the regular BSI genotype.Germination in vivo and in vitro were similar in BSI and Ofer,but pollen tubes failed to reach the ovules in the regular BSIgenotype, while penetrating the ovules in Ofer and the small-ovuledgenotype of BSI. Good-quality fruits similar to Ofer fruitsin weight but with a higher peel to pulp ratio developed inBSI after flower-sterilization in the spring and in the autumn.In Ofer, sterilized flowers failed to develop fruits in springand partially set fruit in the autumn; the fruits consistedalmost exclusively of peel tissue. It was concluded that BSIis a vegetative parthenocarpic clone, i.e., that pollinationis not required for fruit set and development.Copyright 1993,1999 Academic Press Opuntia ficus-indica, cactus pear, fruit development, parthenocarpy, pollen-tube-growth, fluorescence microscopy     

Radial Hydraulic Conductivity of Individual Root Tissues of Opuntia ficus-indica (L.) Miller as Soil Moisture Varies

The constraints on water uptake imposed by individual root tissueswere examined forOpuntia ficus-indicaunder wet, drying, andrewetted soil conditions. Root hydraulic conductivity (L_P) andaxial conductance (K_h) were measured for intact root segmentsfrom the distal region with an endodermis and from midroot witha periderm;L_Pwas then measured for each segment with successivetissues removed by dissection. Radial conductivity (L_R) wascalculated fromL_PandK_hfor the intact segment and for the individualtissues by considering the tissue conductivities in series.Under wet conditions,L_Rfor intact distal root segments was lowestfor the cortex; at midroot, where cortical cells are dead,L_Rforthe cortex was higher and no single tissue was the predominantlimiter ofL_R.L_Rfor the endodermis and the periderm were similarunder wet conditions. During 30d of soil drying,L_Rfor the distalcortex increased almost three-fold due to the death of corticalcells, whereasL_Rfor the midroot cortex was unaffected;L_Rforthe endodermis and the periderm decreased by 40 and 90%, respectively,during drying. For both root regions under wet conditions, thevascular cylinder had the highestL_R, which decreased by 50–70%during 30d of soil drying. After 3d of rewetting, new lateralroots emerged, increasingL_Rfor the tissues outside the vascularcylinder as well as increasing uptake of an apoplastic tracerinto the xylem of both the roots and the shoot. The averageL_Rforintact root segments was similar under wet and rewetted conditions,but the conductivity of the tissues outside the vascular cylinderincreased after rewetting, as did the contribution of the apoplasticpathway to water uptake. Opuntia ficus-indica; prickly pear; root hydraulic conductivity; endodermis; periderm; apoplast; lateral root emergence     

Carbon and water relations for developing fruits of Opuntia ficus-indica (L.) Miller, including effects of drought and gibberellic acid

Growth, gas exchange rates, and carbohydrate content were studied for developing fruits of the cultivated cactus Opuntia ficus-indica (L.) Miller, including effects of drought and exogenous gibberellic acid (GA3). Fruit development required 110 d from the time of bud differentiation to ripening at 80 d after anthesis, when the fruit mass averaged 67 g. Stomatal conductance and net CO2 uptake rates for fruits were higher during the night; they were maximal at 7 d before anthesis and decreased as development progressed. Fruits undergoing drought, imposed by detaching terminal stems bearing fruits, were 50% smaller than the control at 80 d after anthesis and did not ripen. Fruits injected with 2 ml of 500 ppm GA3 were 30% smaller than the control at 80 d after anthesis; they contained a large proportion of aborted seeds that produced a weak sink signal for dry mass accumulation. Gas exchange was higher at 21 d after anthesis for fruits treated with GA3. Total soluble sugars represented 40% of the fruit's dry mass until 45 d after anthesis, when the sugar content rapidly increased, reaching 90% at 73 d after anthesis. Such an increase was not observed for fruits treated with GA3, and the sugar content for fruits undergoing drought remained low throughout development. Starch content increased for developing fruits of O. ficus-indica until 14 d after anthesis and, except for the fruits undergoing drought, decreased thereafter. Fruit development for O. ficus-indica is apparently regulated by water availability as well as hormonal signals originating both within and outside the fruit.     

Past and present role of the Indian-fig prickly-pear (Opuntia ficus-indica (L.) Miller,Cactaceae) in the agriculture of sicily

Of prickly-pear cacti occurring in Sicily, the most widespread and economically important isOpuntia ficus-indica (L.) Miller. In Sicily it has, since its introduction, played an important role in the exploitation of marginal areas. The Sicilian experience is described with reference to the historical outlines and the present intensive production of late fruit. Information on historical and actual uses of the plant and its products (flowers, cladodes, fruits) is given.     

Effect of polysaccharides from Opuntia ficus-indica (L.) cladodes on the healing of dermal wounds in the rat.

In traditional medicine extracts of polysaccharide-containing plants are widely employed for the treatment of skin and epithelium wounds and of mucous membrane irritation. The extracts of Opuntia ficus-indica cladodes are used in folk medicine for their antiulcer and wound-healing activities. The present study describes the wound-healing potential of two lyophilized polysaccharide extracts obtained from O. ficus-indica (L.) cladodes applied on large full-thickness wounds in the rat. When topically applied for 6 days, polysaccharides with a molecular weight (MW)>10(4)Da from O. ficus-indica cladodes induce a beneficial effect on cutaneous repair in this experimental model; in particular the topical application of O. ficus-indica extracts on skin lesions accelerates the reepithelization and remodelling phases, also by affecting cell-matrix interactions and by modulating laminin deposition. Furthermore, the wound-healing effect is more marked for polysaccharides with a MW ranging 10(4)-10(6)Da than for those with MW>10(6)Da, leading us to suppose that the fine structure of these polysaccharides and thus their particular hygroscopic, rheologic and viscoelastic properties may be essential for the wound-healing promoter activity observed.     

A Stereological Analysis of the Succulent Tissue 5Opuntia ficus-indica (L.) Mill.: I. DEVELOPMENT OF MUCILAGE CELLS

A correlated stereological and developmental analysis of thesucculent tissue of Opuntia ficus-indica was carried out. Thestereological parameters related to mucilage cells were describedas a function of their distance from the apical meristem. Polynomesof the second and third degree were found to describe the developmentalprocess. Three zones of development could be distinguished.The relation of the structure of the tissue to its physiologicalfunction and mucilage secretion is discussed.     

A Stereological Analysis of the Succulent Tissue 5Opuntia ficus-indica (L.) Mill.: II. ULTRASTRUCTURAL DEVELOPMENT OF THE MUCILAGE CELLS

Five stages of the development of the cytoplasm of mucilagecells were followed by stereological analysis and analysed togive numerical (N_v), volumetric (V_v) and surface (S_v) densitiesof organelles and membranes. A quantitative evaluation of membrane areas, membrane sites,and mucilage secretion was obtained based on morphological andhistochemical data. The developmental process from early differentiationto complete degeneration of the cytoplasm is discussed in quantitativeterms.     

Diversity of Unavailable Polysaccharides and Dietary Fiber in Domesticated Nopalito and Cactus Pear Fruit (Opuntia spp.)

The aim of this study was to quantify mucilages, pectins, hemicelluloses, and cellulose of nopalitos (edible, as vegetable, young cladodes of flat-stemmed spiny cacti) of most consumed Mexican cultivars, and sweet and acid cactus pear fruits of Opuntia spp. The hypothesis is that, regardless of their unavailable polysaccharides diversity, nopalitos and cactus pear fruits are rich sources of soluble and insoluble dietary fiber. Twelve cultivars of Opuntia spp. were used. Nopalitos had a significant variation in structural polysaccharides among the cultivars: mucilages (from 3.8 to 8.6% dry matter (DM)) averaged near a half of pectins content (from 6.1 to 14.2% DM) and tightly bound hemicelluloses (from 2.2 to 4.7% DM), which were the less abundant polysaccharides, amounted 50% of the loosely bound hemicelluloses (from 4.3 to 10.7% DM). Acid fruits (or 'xoconostle') had significantly higher unavailable polysaccharides content than sweet fruit, and contain similar proportions than nopalitos. Unavailable polysaccharides represent a high proportion of dry tissues of nopalitos and cactus pear fruits, composition of both of these soluble and insoluble polysaccharides (total dietary fiber) widely vary among cultivars without an evident pattern. Nopalitos and cactus pear fruit can be considered an excellent source of dietary fiber.     

Anti-Inflammatory and Antinociceptive Properties of Flavonoids from the Fruits of Black Mulberry (Morus nigra L.)

We analyzed the anti-inflammatory and antinociceptive activities of total flavonoids (TF) found in black mulberry fruits. The TF content was 20.9 mg/g (dry weight). Two anthocyanins, cyanidin-3-O-glucoside (8.3 mg/g) and cyanidin-3-O-rutinoside (2.9 mg/g), were identified in the fruits by UPLC. The TF of black mulberry fruits had significant reducing power and radical (OH^-,O2.−, DPPH and ABTS) scavenging activities that was demonstrated in a dose-response curve. The TF had inhibitory activities on xylene-induced ear edema and carrageenan-induced paw edema in mice. In addition, TF had antinociceptive activities in the two nociceptive phases of formalin test. We used ELISA to detect the pro-inflammatory cytokines IL-1β, TNF-α, IFN-γ, and NO in the serum of mice. These cytokines were significantly inhibited or scavenged by TF (50 and 100 mg/kg). The results demonstrated that TF of black mulberry possess anti-inflammatory and analgesic effects that might correlate to its antioxidant activities and inhibition of pro-inflammatory cytokines.     

Correction to: The Influence of Light Wavelength on Growth and Antioxidant Capacity in Pachyrhizus erosus (L.) Urban

Journal of Plant Growth Regulation - The original version of this article unfortunately contained an error in Acknowledgement. The authors would like to correct the error with this erratum. The...     

Flavonoid content in leaf extracts of the fig (Ficus carica L.), carob (Ceratonia siliqua L.) and pistachio (Pistacia lentiscus L.)

The total flavonoid content of leaf extracts (70% ethanol) from fig (Ficus carica L.), carob (Ceratonia siliqua L.) and pistachio (Pistacia lentiscus L.) plants were determined by using reverse phase high-performance liquid chromatography (HPLC)-and analyzed by UV/VIS array and electrospray ionization (ESI)-mass spectrometry (MS) detectors. As a base for comparison, flavonoid type and level were also determined in extracts from soybeans and grape seeds. It was found that the major flavonoids in Ficus are quercetin and luteolin, with a total of 631 and 681 mg/kg extract, respectively. In Ceratonia leaves, nine different flavonoids were detected. The major one was myricetin (1486 mg/kg extract), with a similar level in Pistacia (1331 mg/kg extract, myricetin). The present study is the first to report the presence of the isoflavone genistein in the Pistacia leaf, which was discovered to consist of about a third of the genistein level detected in soybean.     

Antioxidant Properties and Flavonoid Profile in Leaves of Calabrian Lavandula multifida L., an Autochthon Plant of Mediterranean Southern Regions

Lavandula multifida is a rare short‐lived plant characteristic of Mediterranean basin able to survive in hot and arid climatic conditions on poorly evolved limestone soils. In this work, we characterize the enzymatic antioxidant system and phenolic composition, as well as the antioxidant properties of L. multifida fresh leaves. Enzymatic patterns show high level of peroxidases, ascorbate peroxidase, and dehydroascorbate reductase activities, when compared with L. angustifolia. The same trend is evident in total carotenoids, ascorbic acid, and reduced glutathione, and in the total antioxidant capacity assay. Moreover, RP‐DAD‐HPLC analyses of EtOH extract, obtained from fresh leaves, reveal main components, carvacrol, vitexin, and 7‐ or 8‐glucoside derivatives of hypolaetin, scutellarein, luteolin, isoscutellarein, apigenin, and chrysoeriol. The analysis of this autochthon plant depicted a series of strategies adopted by L. multifida to survive in its stressful natural habitat and richness in health‐promoting compounds that can be a resource for the preservation of this variety in dangerous of extinction.     

Antioxidant properties of prunes (Prunus domestica L.) and their constituents

Prunes contain large amounts of phenolics and show high antioxidant activity. The aim of this study is to clarify the contents of caffeoylquinic acid (CQA) isomers, and to estimate the contribution of these isomers to the antioxidant activity of prunes. Furthermore, structural elucidation and evaluation of antioxidant activity of prune components were also performed. CQA isomers in prunes were quantified by HPLC analysis, and it has become apparent that prunes contain relatively high amount of 4-O-caffeoylquinic acid. The contribution of CQA isomers to the antioxidant activity of prunes was revealed to be 28.4% on the basis of oxygen radical absorbance capacity (ORAC); hence, it was indicated that residual ORAC is dependent on unknown antioxidant components. Total 28 compounds were isolated and their structures were elucidated by NMR and MS analyses. Four abscisic acid related compounds, a chromanon, and a bipyrrole were novel. Each CQA isomer in prunes showed high antioxidant activities when measured by the oil stability index (OSI) method, O2- scavenging activity, and ORAC. Other isolated compounds such as hydroxycinnamic acids, benzoic acids, coumarins, lignans, and flavonoid showed high ORAC values. Furthermore, a novel chromanon indicated a remarkable synergistic effect on ORAC of CQA isomers.     

Surface of Lactic Acid Bacteria: Relationships between Chemical Composition and Physicochemical Properties

The surface chemical composition and physicochemical properties (hydrophobicity and zeta potential) of two lactic acid bacteria, Lactococcus lactis subsp. lactis bv. diacetilactis and Lactobacillus helveticus, have been investigated using cells harvested in exponential or stationary growth phase. The surface composition determined by X-ray photoelectron spectroscopy (XPS) was converted into a molecular composition in terms of proteins, polysaccharides, and hydrocarbonlike compounds. The concentration of the last was always below 15% (wt/wt), which is related to the hydrophilic character revealed by water contact angles of less than 30°. The surfaces of L. lactis cells had a polysaccharide concentration about twice that of proteins. The S-layer of L. helveticus was either interrupted or crossed by polysaccharide-rich compounds; the concentration of the latter was higher in the stationary growth phase than in the exponential growth phase. Further progress was made in the interpretation of XPS data in terms of chemical functions by showing that the oxygen component at 531.2 eV contains a contribution of phosphate in addition to the main contribution of the peptide link. The isoelectric points were around 2 and 3, and the electrophoretic mobilities above pH 5 (ionic strength, 1 mM) were about −3.0 × 10⁻⁸ and −0.6 × 10⁻⁸ m² s⁻¹ V⁻¹ for L. lactis and L. helveticus, respectively. The electrokinetic properties of the latter reveal the influence of carboxyl groups, while the difference between the two strains is related to a difference between N/P surface concentration ratios, reflecting the relative exposure of proteins and phosphate groups at the surface.     

Lactic acid bacteria associated with the digestive tract of Atlantic salmon (Salmo salar L.)

The present study reports the effect of excessive handling stress and starvation on the lactic acid bacteria associated with the digestive tract of Atlantic salmon (Salmo salar L.). A relatively low population level (approximately 2 x 103 bacteria per gram wet tissue) of viable adherent heterotrophic bacteria was associated with the digestive tract (foregut, midgut and hindgut). Of the 752 bacterial isolates isolated from diet, water and the digestive tract, 201 isolates belonged to the carnobacteria. Of these isolates, one from the diet, one from the rearing water and 80 from the gastrointestinal tract, were further identified on the basis of 16S rDNA sequence analysis. All these isolates were identified as being Carnobacterium piscicola-like. Daily repeated stress and starvation of the fish over 11 d had no influence on the total culturable bacterial numbers or population level of C. piscicola associated with the digestive tract. C. piscicola-like isolates colonizing the various intestinal regions (foregut, midgut and hindgut) were also screened for their ability to produce growth inhibitory compounds active against the fish pathogen Aeromonas salmonicida. Of the 199 C. piscicola isolates tested, 139 inhibited growth of the pathogen.     

Liposomal Drug with Carnosine and Lipoic Acid: Preparation,Antioxidant and Antiplatelet Properties

The conditions for producing phosphatidylcholine liposomes containing lipoic acid and carnosine together were determined. The obtained liposomes are 180–250-nm spherical particles with an efficiency of lipoic acid inclusion of 50–70% (for carnosine, 17–33%). Based on the model of the oxidation of phosphatidylcholine by hydrogen peroxide, an antioxidant effect of carnosine, lipoic acid or lipoic acid with carnosine together was demonstrated; it consisted in inhibition of lipid peroxidation process, which was manifested in a decrease in the formation of lipid peroxidation products that react with thiobarbituric acid. It was established that lipoic acid (5 mM) and carnosine (0.1–10 mM) in liposomes exhibit an antioxidant effect. At the same time, it was demonstrated that the content of the appropriate lipid peroxidation products in liposomes with antioxidants (lipoic acid + carnosine) was 15 times lower than in control liposomes (without antioxidants). The effect of the obtained liposomal drugs on the platelet aggregation induced by arachidonic acid was evaluated. It was found that the liposomal drug containing lipoic acid (1.5 mM) and carnosine (2.1 mM) inhibited platelet aggregation by 50–55% relative to the control (platelets and arachidonic acid), while liposomes without antioxidants and water-soluble forms of carnosine and lipoic acid had almost no effect on platelet aggregation caused by arachidonic acid.

     

Thermotolerance and Related Antioxidant Enzyme Activities Induced by Heat Acclimation and Salicylic Acid in Grape (Vitis vinifera L.) Leaves

Thermotolerance and related antioxidant enzyme activities induced by both heat acclimation and exogenous salicylic acid (SA) application were studied in grapevine (Vitis vinifera L. cv. Jingxiu). Heat acclimation and exogenous SA application induced comparable changes in thermotolerance, ascorbic acid (AsA), glutathione (GSH), and hydrogen peroxide (H₂O₂) concentrations, and in activities of the antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD), glutathione reductase (GR), ascorbic peroxidase (APX) and catalase (CAT) in grape leaves. Within 1 h at 38 °C, free SA concentration in leaves rose from 3.1 μg g⁻¹ FW to 19.1 μg g⁻¹ FW, then sharply declined. SA application and heat acclimation induced thermotolerance were related to changes of antioxidant enzyme activities and antioxidant concentration, indicating a role for endogenous SA in heat acclimation in grape leaves.     

Properties and Characteristics of Multi-Layered Films from Tilapia Skin Gelatin and Poly(Lactic Acid)

Multi-layered films based on tilapia skin gelatin and poly(lactic acid) (PLA) were characterized, in comparison with the control gelatin and PLA films. Three different layers of multi-layered films (PLA/Gelatin/PLA) were visualized by scanning electron microscopic (SEM) analysis. The synergetic effect of lamination was evidenced by the increased mechanical properties (P < 0.05). Multi-layered films had higher water vapor barrier property and water resistance, compared to control gelatin film (P < 0.05). Gelatin films showed increased lightness (L*) with coincidental decrease in total color difference (?E*) in the presence of PLA layers (P < 0.05). Transparency and solubility of films decreased with increasing ratio of PLA (P < 0.05). In addition, multi-layered films showed the enhanced hydrophobicity and thermal stability as evidenced by increased water contact angle and degradation temperature, respectively. Thus, PLA/Gelatin/PLA multi-layered film with improved water vapour barrier property could serve as bio-degradable packaging material for wider applications.