The antihyperuricemia activity of Astragali Radix through regulating the expression of uric acid transporters via PI3K/Akt signalling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Astragali Radix (AR) is the dry root of the leguminous plants Astragalus membranaceus (Fisch) Beg. var. mongholicus (Beg) Hsiao, and Astragalus membranaceus (Fisch) Bge., being used as a medicinal and edible resource. AR is used in traditional Chinese medicine prescriptions to treat hyperuricemia, but this particular effect is rarely reported, and the associated mechanism of action is still need to be elucidated. AIM OF THE STUDY: To research the uric acid (UA)-lowering activity and mechanism of AR and the representative compounds through the constructed hyperuricemia mouse and cellular models. MATERIALS AND METHODS: In our study, the chemical profile of AR was analysed by UHPLC-QE-MS, as well as the mechanism of action of AR and the representative compounds on hyperuricemia was studied through the constructed hyperuricemia mouse and cellular models. RESULTS: The main compounds in AR were terpenoids, flavonoids and alkaloids. Mice group treated with the highest AR dosage showed significantly lower (p < 0.0001) serum uric acid (208 ± 9 µmol/L) than the control group (317 ± 11 µmol/L). Furthermore, UA increased in a dose-dependence manner in urine and faeces. Serum creatinine and blood urea nitrogen standards, as well as xanthine oxidase in mice liver, decreased (p < 0.05) in all cases, indicating that AR could relieve acute hyperuricemia. UA reabsorption protein (URAT1 and GLUT9) was down-regulated in AR administration groups, while the secretory protein (ABCG2) was up-regulated, indicating that AR could promote the excretion of UA by regulating UA transporters via PI3K/Akt signalling pathway. CONCLUSION: This study validated the activity, and revealed the mechanism of AR in reducing UA, which provided experimental and clinical basis for the treatment of hyperuricemia with it.

Water-in-Oil-in-Water Double Emulsions as Protective Carriers for Sambucus nigra L. Coloring Systems.

The use of natural colorants is needed to overcome consumer concerns regarding synthetic food colorants' safety. However, natural pigments have, in general, poor stability against environmental stresses such as temperature, ionic strength, moisture, light, and pH, among others. In this work, water-in-oil-in-water (W1/O/W2) emulsions were used as protective carriers to improve color stability of a hydrophilic Sambucus nigra L. extract against pH changes. The chemical system comprised water and corn oil as the aqueous and oil phases, respectively, and polyglycerol polyricinoleate (PGPR), Tween 80, and gum Arabic as stabilizers. The primary emulsion was prepared using a W1/O ratio of 40/60 (v/v). For the secondary emulsion, W1/O/W2, different (W1/O)/W2 ratios were tested with the 50/50 (v/v) formulation presenting the best stability, being selected as the coloring system to test in food matrices of different pH: natural yogurt (pH 4.65), rice drink (pH 6.01), cow milk (pH 6.47), and soy drink (pH 7.92). Compared to the direct use of the extract, the double emulsion solution gave rise to higher color stability with pH change and storage time, as corroborated by visual and statistical analysis.

Valorization of Bio-Residues from the Processing of Main Portuguese Fruit Crops: From Discarded Waste to Health Promoting Compounds.

Food processing generates a large amount of bio-residues, which have become the focus of different studies aimed at valorizing this low-cost source of bioactive compounds. High fruit consumption is associated with beneficial health effects and, therefore, bio-waste and its constituents arouse therapeutic interest. The present work focuses on the main Portuguese fruit crops and revises (i) the chemical constituents of apple, orange, and pear pomace as potential sources of functional/bioactive compounds; (ii) the bioactive evidence and potential therapeutic use of bio-waste generated in the processing of the main Portuguese fruit crops; and (iii) potential applications in the food, nutraceutical, pharmaceutical, and cosmetics industries. The current evidence of the effect of these bio-residues as antioxidant, anti-inflammatory, and antimicrobial agents is also summarized. Conclusions of the revised data are that these bio-wastes hold great potential to be employed in specific nutritional and pharmaceutical applications.

Potato biodiversity: A linear discriminant analysis on the nutritional and physicochemical composition of fifty genotypes.

Fifty potato genotypes from twenty-four different countries of origin, four different flesh colours (yellow, purple, red and marble) and different cultivation types (Andean accessions, landraces, breeder lines and cultivated varieties) were studied in terms of their nutritional and physicochemical characteristics. In general, cultivated varieties and breeder lines showed the highest similarity (slight differences only in some particular fatty acids distributions: C10:0, C12:0 and C22:0) concerning the physicochemical parameters assayed in this work, independently of the geographical origin or tuber flesh colour of these genotypes. Nonetheless, some of the studied landraces and Andean accessions proved to be similar enough to be considered as genotypes with good potential for commercial cultivation. These results can contribute to the supply of new potato genotypes into sustainable farming systems, supporting the protection of potato biodiversity, particularly Andean accessions, landraces and coloured genotypes (red or purple flesh) which are not widely cultivated so far.

Ficus carica L. and Prunus spinosa L. extracts as new anthocyanin-based food colorants: A thorough study in confectionery products.

The present work describes the evaluation of fig peels and blackthorn fruit extracts as natural purple colorants in doughnuts (icing) and in a typical Brazilian pastry called "beijinho". The extracts were screened for their antioxidant activity as well as their antibacterial capacity. Nutritionally, the employed extracts did not induce significant changes, contrarily to the observed for the rheological features, mainly the darker purple tone observed when blackthorn extract was used in the icing solution. After 24 h, both prepared formulations showed a decrease in color intensity, with no significant differences between fig and blackthorn extracts. In turn, the firmness and consistency of the doughnuts benefited from using natural colorants in the icing solution, while "beijinhos" became softer and chewier, which are valued attributes. A significant increase in the antioxidant and antimicrobial activities was also observed for both natural extracts. Accordingly, the evaluated extracts are promising candidates as natural food colorants.

Phenolic Profile of Croton urucurana Baill. Leaves, Stems and Bark: Pairwise Influence of Drying Temperature and Extraction Solvent.

Cerrado biome represents an area with great biodiversity. Some of its plants have significant ethnopharmacological uses, with specific purposes. Croton urucurana Baill., for instance, was previously acknowledged for its anti-hemorrhagic, anti-inflammatory, antiseptic, healing, and potentially antifungal and entomopathogenic actions. Nevertheless, the compounds supporting these empirical applications are still unknown. Accordingly, this work was designed to achieve a complete characterization of the phenolic profile of different botanical tissues obtained from C. urucurana, and also to verify how different operational conditions (different drying temperatures and extraction conditions) affect that profile. All samples were further characterized by HPLC-DAD-ESI/MSn, and results were compared by advanced chemometric tools. In general, the drying temperatures that maximize the extraction yield of specific individual phenolic compounds were established. Likewise, it was possible to verify that samples extracted with the hydroethanolic solution allowed higher phenolic yields, either in individual compounds (except (epi)catechin-di-O-gallate) or total phenolics. The identification of the best operational conditions and phenolic profiles associated with each C. urucurana botanical part contributes to enabling their use in food or pharmaceutical-related applications.

Characterization and Application of Pomegranate Epicarp Extracts as Functional Ingredients in a Typical Brazilian Pastry Product.

Currently, there is a clear tendency to incorporate natural ingredients into food and pharmaceutical formulations. Besides being well-accepted by consumers, these ingredients have less adverse side effects than their artificial counterparts. The pomegranate processing industry produces large quantities of by-products that are discarded as bio-residues, despite containing bioactive compounds. Accordingly, the epicarp of two pomegranate varieties (Mollar de Elche and Purple Queen) was tested as a potential source of bioactive compounds with food application. The phenolic profile was identified by HPLC-DAD-ESI/MS, revealing fourteen phenolic compounds in both varieties (Purple Queen showed also three anthocyanins), with punicalagin isomers as the major compounds. Nonetheless, Mollar de Elche presented greater antioxidant and antibacterial activities. Despite this result, Purple Queen was selected to be tested as a new natural colouring and functionalizing ingredient in a Brazilian pastry product. The incorporation of the selected extract maintained the nutritional profile and provided a higher antioxidant activity compared to the traditional product. In this way, this work confirmed the possible use of pomegranate epicarp as a natural ingredient in the food industry, conferring dyeing and functionalizing effects, and anticipating a possible valorisation of this bio-residue.

Extracts from Vaccinium myrtillus L. fruits as a source of natural colorants: chemical characterization and incorporation in yogurts.

The food industry is always seeking innovative approaches to maintain consumers' interest and increase their awareness towards the healthiness of diets. Therefore, much interest has been given to natural food additives, namely colourants. In this work, a bilberry extract was primarily characterized in terms of anthocyanin compounds by UPLC-DAD-ESI/MSn, and its colouring capacity was further compared with a synthetic anthocyanin colourant (E163) in different yogurt formulations. The prepared samples were evaluated in different periods (0 and 7 days) for nutritional profile, individual fatty acids, soluble sugars and external colour to determine the effects of each additive. Overall, the major anthocyanin compounds in bilberry were malvidin glycoside and delphinidin glycoside derivatives. With regard to the prepared yogurts, all samples maintained the nutritional profile, individual fatty acids and soluble sugars, independent of storage time. Nonetheless, bilberry extracts showed lower colouring capacity when compared to that of E163, despite the higher stability of bilberry extract throughout the storage time, when compared to that of E163. Furthermore, another advantage of this natural extract is that it has potential bioactive properties that can be conferred to foods, due to their high content of bioactive compounds, such as anthocyanins.

Castanea sativa male flower extracts as an alternative additive in the Portuguese pastry delicacy "pastel de nata".

Replacing artificial additives by natural compounds is a current trend in the food industry. In addition to their preserving effect, naturally obtained ingredients often exhibit important levels of bioactivities. Generally, plant species represent better sources of natural ingredients, since their compounds are less prone to causing unpleasant taste or odour. Chestnut male flower (CMF) was reported to have high antioxidant and antimicrobial activities. Hence, it was tested as an alternative to potassium sorbate in the most treasured Portuguese delicacy: "pastel de nata". Different nutritional, chemical, physical and bioactive parameters were compared in two different periods: baking day and two days after baking. All samples presented similar nutritional and chemical profiles, but those added with CMF revealed higher contents of reducing agents and radical scavengers. Accordingly, the newly obtained formulation is expected to have better effects on consumers' health, maintaining the chemical characteristics, besides rendering a novel, economically profitable, application to CMF.

Incorporation of tocopherol-rich extracts from mushroom mycelia into yogurt.

Consumers are well-informed about food additives and it is likely that they prefer natural additives over their synthetic analogues. Antioxidants represent a major class of food preservatives, among which tocopherols stand out as one of the most important examples. Interestingly, these compounds are present in relevant amounts in the mycelia of in vitro cultured mushrooms. Accordingly, the mycelia from Ganoderma lucidum, Pleurotus ostreatus and Pleurotus eryngii were used as alternative sources of tocopherols. These extracts were incorporated into different yogurt formulations, which were further compared among each other and with yogurts containing commercial α-tocopherol (E307), regarding their nutritional parameters, fatty acid profile and antioxidant activity. The proposed approach was validated as an effective functionalization strategy, particularly in the case of the G. lucidum mycelium, which showed the highest antioxidant potential, most likely as a result of its tocopherol profile. Furthermore, yogurts prepared with each mycelium extract allowed maintaining the nutritional properties observed in the "blank" yogurt formulation.

Suitability of lemon balm (Melissa officinalis L.) extract rich in rosmarinic acid as a potential enhancer of functional properties in cupcakes.

Melissa officinalis (lemon balm) and its extracts have been frequently reported as possessing bioactive properties, offering the potential for use in development/enrichment of food products with additional functional capabilities, providing health benefits to consumers. The antioxidant, antibacterial and antifungal activity of lemon balm extract, as well as its potential hepatotoxicity were thoroughly evaluated. The extracts were then incorporated into cupcakes and their preserving effect, chemical composition, colour parameters and antioxidant activity were compared with those provided by potassium sorbate. In general, the variables with the largest differences among different storage times were energy level, sucrose, glucose, palmitic acid (C6:0) and oleic acid (C18:1n9). On the other hand, L∗ (top), a∗ (top), b∗ (top), pH, capric acid (C10:0) and lauric acid (C12:0) showed the greatest variation according to cupcake formulation. The results observed indicate that the lemon balm extract rich in rosmarinic acid can provide advantageous functional properties to bakery products.

Bioactivity and chemical characterization of Opuntia macrorhiza Engelm. seed oil: potential food and pharmaceutical applications.

In the food industry, there is a continuous search for ingredients that might provide advantageous properties to food products, either considering their nutritional value or bioactivity, as well as flavouring and technological aspects. Crude oils are good examples of this type of ingredient, especially if obtained from nonconventional sources. Accordingly, the Opuntia macrorhiza Engelm. seed oil (OMSO) was chemically characterized and evaluated for different in vitro and in vivo bioactivities. OMSO presented physicochemical characteristics appropriate to be considered as an edible oil, namely low acidity value, stability to oxidation (high peroxide value and low K232 and K270 values), and high contents of unsaturated fatty acids (as shown by the iodine value) and saponifiable matter. Furthermore, this natural oil, owing to its rich phytochemical profile, showed relevant antioxidant activity (especially in lipid peroxidation inhibition assays), α-glucosidase inhibitory activity, cytotoxicity against human tumour cell lines, antibacterial (mainly against Gram positive species) and antifungal properties, as well as anti-inflammatory and analgesic activities. Furthermore, OMSO did not show any sign of acute toxicity on animals, highlighting its possible use in different applications, considering that this natural product is not expected to induce the adverse effects typically associated with synthetic bioactive agents (e.g., ampicillin, amphotericin B, or lysine acetylsalicilate).

Evaluation of the cytotoxicity (HepG2) and chemical composition of polar extracts from the ruderal species Coleostephus myconis (L.) Rchb.f.

Coleostephus myconis (L.) Rchb.f. (Asteraceae) is a highly disseminated plant species with ruderal and persistent growth. Owing to its advantageous agronomic properties, C. myconis might have industrial applications. However, this species needs to be comprehensively characterized before any potential use. In a previous study, the phenolic composition and antioxidant activity of different C. myconis tissues were characterized. This investigation was extended to examine the cytotoxic potential of selected plant tissues (flowers and green parts) using a HepG2 cell line by utilizing the lysosomal neutral red uptake assay or mitochondrial (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay. In addition, the macronutrients content, lipophilic compounds (fatty acids, tocopherols), and amino acids were also determined. C. myconis flowers were used in the senescence stage, which was previously identified as the stage that presented maximal phenolic content and highest antioxidant activity. In contrast, stems and leaves were employed due to their high biomass proportion. Regarding cytotoxicity, mitochondrial and lysosomal damage was only significant when HepG2 cells were exposed to the highest extract concentrations (stems and leaves, 0.9 mg/ml; senescent flowers, 0.3 mg/ml). Chemically, the senescent flowers were mostly characterized by their high levels of fat, amino acids (especially threonine), oleic acid, ß-, and γ-tocopherol, while stems and leaves contained high concentrations of carbohydrates, linolenic acid, and α-tocopherol. In general, these results provide information regarding the threshold concentrations of C. myconis extracts that might be used in different applications without toxicity hazards.

Chemical Profiling and Assessment of Antineurodegenerative and Antioxidant Properties of Veronica teucrium L. and Veronica jacquinii Baumg.

Neuroprotective potential of V. teucrium and V. jacquinii methanol extracts was analyzed. Chemical analysis of investigated extracts showed the presence of phenolic acid derivatives, flavonoids and one secoiridoid. The detected flavonoids derived from flavones (luteolin and isoscutellarein in V. jacquinii; apigenin, isoscutellarein and luteolin in V. teucrium) and flavonol (quercetin in V. jacquinii). Acteoside was the dominant compound in V. jacquinii, while plantamajoside and isoscutellarein 7-O-(6‴-O-acetyl)-ß-allosyl (1‴→2‴)-ß-glucoside were the major phenolics in V. teucrium. Additionally, the antineurodegenerative activity was tested at concentrations of 25, 50, and 100 µg/ml using acetylcholinesterase (AChE) and tyrosinase (TYR) assays. The inhibition of both enzymes was achieved with the investigated extracts, ranging from 22.78 to 35.40% for AChE and from 9.57 to 16.38% for TYR. There was no statistical difference between the activities of the analyzed extracts. Our data indicate that V. teucrium and V. jacquinii may have beneficial effects against Alzheimer's and Parkinson's disease.

Chestnut and lemon balm based ingredients as natural preserving agents of the nutritional profile in matured "Serra da Estrela" cheese.

Chestnut flowers, lemon balm plants and their decoctions were incorporated into "Serra da Estrela" cheese, to assess their potential to preserve its nutritional properties and provide new foodstuffs. The analyses were carried out after the normal ripening period of 1month and after 6months of storage. The most abundant nutrients were proteins and fats. The most abundant minerals were Ca and Na, while C16:0 and C18:1 were the main fatty acids. Saturated fatty acids were the most abundant, followed by the monounsaturated. Moisture seemed to be lower in the samples with the plants incorporated. The dried plants, when incorporated, seemed to be more efficient as preservers then the decoctions, although these better preserved the proteins. These plants can be regarded as promising natural preservers in foodstuffs cheese, given the preservation of key parameters and the slight impact on the nutritional value.

Bioactivity, proximate, mineral and volatile profiles along the flowering stages of Opuntia microdasys (Lehm.): defining potential applications.

Opuntia spp. flowers have been traditionally used for medical purposes, mostly because of their diversity in bioactive molecules with health promoting properties. The proximate, mineral and volatile compound profiles, together with the cytotoxic and antimicrobial properties were characterized in O. microdasys flowers at different maturity stages, revealing several statistically significant differences. O. microdasys stood out mainly for its high contents of dietary fiber, potassium and camphor, and its high activities against HCT15 cells, Staphylococcus aureus, Aspergillus versicolor and Penicillium funiculosum. The vegetative stage showed the highest cytotoxic and antifungal activities, whilst the full flowering stage was particularly active against bacterial species. The complete dataset has been classified by principal component analysis, achieving clearly identifiable groups for each flowering stage, elucidating also the most distinctive features, and comprehensively profiling each of the assayed stages. The results might be useful to define the best flowering stage considering practical application purposes.

Cottage cheeses functionalized with fennel and chamomile extracts: Comparative performance between free and microencapsulated forms.

Globally, there is a trend for healthy food products, preferably incorporating natural bioactive ingredients, replacing synthetic additives. From previous screening studies, extracts of Foeniculum vulgare Mill. (fennel) and Matricaria recutita L. (chamomile) maintained nutritional properties and improved the antioxidant activity of cottage cheese. Nevertheless, this effect was limited to 7 days. Accordingly, aqueous extracts of these plants were microencapsulated in alginate and incorporated into cottage cheese to achieve an extended bioactivity. Plain cottage cheese, and cheese functionalized by direct addition of free decoctions, were prepared and compared. Independently of plant species, "functionalization type" factor did not show a significant effect on the nutritional parameters, as also confirmed in the linear discriminant analysis, where these parameters were not selected as discriminating variables. Furthermore, samples functionalized with microencapsulated extracts showed higher antioxidant activity after the 7th day, thereby demonstrating that the main purpose of this experimental work was achieved.

Medicago spp. as potential sources of bioactive isoflavones: Characterization according to phylogenetic and phenologic factors.

A high variety of plant species are often proposed as potential natural sources of specific bioactive components, with emphasis in phenolic compounds. However, the ability to produce a determined phytochemical might be variable, even among species with close phylogeny. Furthermore, the metabolic dynamics vary greatly according to phenologic factors. Herein, it was verified whether isoflavone production in Medicago spp. is more associated with phylogenetic or phenologic determinants, to define the optimal productive conditions. Isoflavone profiles were characterized in field-grown Medicago species in three phenologic stages. Isoflavones were extracted by matrix solid-phase dispersion method and analyzed using high-performance liquid chromatography coupled with a diode-array detector. The obtained data were evaluated by a generalized linear model (GLM) and linear discriminant analysis (LDA). Formononetin, genistein and irilone were the most abundant isoflavones, reaching values higher than those present in acknowledged plant sources like soy or red clover. Outputs from GLM and LDA indicate that the phylogenetic factors are the most defining criteria. This study promotes Medicago spp. as potential isoflavone sources, particularly because the effects of these compounds are highly dependent on their type and concentration, with potential application as foodstuff, feedstuff, or in the nutraceutical and pharmaceutical industry.

Is honey able to potentiate the antioxidant and cytotoxic properties of medicinal plants consumed as infusions for hepatoprotective effects?

Due to the enormous variety of phytochemicals present in plants, their extracts have been used for centuries in the treatment of innumerable diseases, being perceived as an invaluable source of medicines for humans. Furthermore, the combination of different plants was reported as inducing an improved effect (synergism) in comparison with the additive activity of the plants present in those mixtures. Nevertheless, information regarding the effects of plant infusions added with honey is still rather scarce. Accordingly, the aim of this study was to evaluate the interaction between chestnut honey, a natural product with well-reported beneficial properties, and three medicinal plants (either as a single plant or as combinations of two and three plants), with regard to their antioxidant activity and hepatotoxicity. Antioxidant activity was evaluated by comparing the results from four different assays; hepatotoxicity was assessed in two different cell lines. Results were compared by analysis of variance and linear discriminant analysis. The addition of honey to the infusions had a beneficial result in both cases, producing a synergistic effect in all samples, except ß-carotene bleaching inhibition for artichoke + milk thistle + honey preparation and also preparations with lower hepatotoxicity, except in the case of artichoke + honey. Moreover, from the discriminant linear analysis output, it became obvious that the effect of honey addition overcame that resulting from using single plant or mixed plant based infusions. Also, the enhanced antioxidant activity of infusions containing honey was confirmed by lower hepatotoxicity.

Edible flowers of Viola tricolor L. as a new functional food: antioxidant activity, individual phenolics and effects of gamma and electron-beam irradiation.

Edible flowers are used in food preparations, being also recognized for their beneficial effects on human health. Nevertheless, these species are highly perishable, and irradiation treatment might be applied to ensure food quality and increase their shelf life. Viola tricolor L. is a typical edible flower, with multiple applications and biological properties, mainly provided by the flavonoid content. In the present work, the phenolic compounds were analyzed by HPLC-DAD-ESI/MS, and the antioxidant activity was evaluated using biochemical assays. Linear discriminant analyses (LDA) were performed in order to compare the results obtained with flowers submitted to different irradiation doses and technologies (cobalt-60 and electron-beam). In general, irradiated samples (mostly with 1 kGy) showed the highest phenolic content and antioxidant activity. Furthermore, the significant differences observed in the LDA allow determination of which dose and/or technology is suitable to obtain flowers with higher antioxidant potential.