Protective effect of Bougainvillea glabra Choisy bract in toxicity induced by Paraquat in Drosophila melanogaster.

Paraquat (PQ) is a herbicide widely used in agriculture to control weeds. The damage caused to health through intoxication requires studies to combating its damage to health. Bougainvillea glabra Choisy is a plant native to South America and its bracts contain a variety of compounds, including betalains and phenolic compounds, which have been underexplored about their potential applications and benefits for biological studies to neutralize toxicity. In this study, we evaluated the antioxidant and protective potential of the B. glabra bracts (BBGCE) hydroalcoholic extract against Paraquat-induced toxicity in Drosophila melanogaster. BBGCE demonstrated high antioxidant capacity in vitro through the assays of ferric-reducing antioxidant power (FRAP), radical 2,2-diphenyl-1-picrylhydrazyl (DPPH), free radical ABTS and quantification of phenolic compounds, confirmed through identifying the main compounds. Wild males of D. melanogaster were exposed to Paraquat (1.75 mM) and B. glabra Choisy (1, 10, 50 and 100 µg/mL) in agar medium for 4 days. Flies exposed to Paraquat showed a reduction in survival rate and a significant decrease in climbing capacity and balance test when compared to the control group. Exposure of the flies to Paraquat caused a reduction in acetylcholinesterase activity, an increase in lipid peroxidation and production of reactive species, and a change in the activity of the antioxidant enzymes. Co-exposure with BBGCE was able to block toxicity induced by PQ exposure. Our results demonstrate that bract extract has a protective effect against PQ on the head and body of flies, attenuating behavioral deficit, exerting antioxidant effects and blocking oxidative damage in D. melanogaster.

Gut Microbiota Modulation by Bioactive Compounds from Ilex paraguariensis: an In Vivo Study.

Yerba-mate (Ilex paraguariensis) is recognized for its biocompounds and bioactive properties. This study aimed to assess the potential of yerba-mate extract to modulate the intestinal microbiota in rats. After the ethical committee approval (CEUA - UPF, number 025/2018), the Wistar rats were given a daily dose of 3.29 mg of phenolic compounds per animal for 45 days. The antioxidant activity of the extract was assessed by ABTS and FRAP assays and the total phenolic compounds was measured at different pH levels. Identification and quantification of chlorogenic acid isomers were carried out using high-performance liquid chromatography (HPLC). Intestinal microbiota modulation was evaluated by administering the yerba-mate extract or water (control) to Wistar rats via intragastric gavage and its efficiency was measured through PCR. The antioxidant capacity of the yerba-mate extract was 64.53 ± 0.26 µmol Trolox/mL (ABTS) and 52.96 ± 0.86 µmol Trolox/mL (FRAP). The total phenolic compounds showed higher levels at pH 7.5 compared to pH 2.0. Chlorogenic acid isomers were found in greater abundance, with a concentration of 14.22 g/100 g. The administration of the extract resulted in positive modulation of the intestinal microbiota, specifically for the genera Lactobacillus sp. and Prevotella sp. The increase of these genera is related to the promotion of homeostasis of the gut microbiota. Therefore, these findings indicate that yerba-mate extract possesses significant antioxidant activity and can effectively modulate the intestinal microbiota in rats. These results support the potential use of yerba-mate as an alternative for controlling and preventing diseases associated with intestinal dysbiosis.

Influence of sprouting on the bioaccessibility and bioactivity of benzoxazinoids, phenolic acids, and flavonoids of soft and hard wheat cultivars.

Grain germination increases the contents of benzoxazinoids and the antioxidant capacity of wheat and differentially affects the phytochemical composition of hard and soft wheat cultivars. It was investigated whether wheat cultivars (sprouted or not) with distinct grain texture characteristics (BRS Guaraim, soft vs. BRS Marcante, hard texture) would have different behavior in relation to changes in phytochemical compounds, bioaccessibility and antioxidant capacity during simulated gastrointestinal digestion of a tabbouleh preparation. Sprouting increased the nominal amount of phytochemicals in tabbouleh resulting in increased release of phenolic acids (up to 7.5-fold) and benzoxazinoids (up to 12.5-fold) during all digestion phases besides higher bioaccessibility (up to 2.8-fold). Sprouting caused greater increase in the bioaccessibility of phenolic acids for the soft wheat cultivar (4.5-fold) than for the hard cultivar (1.9-fold) and it increased the colon available index of phenolic acids only for the soft cultivar (1.8-fold). Flavonoids, mainly represented by apigenin glycosides, were marginally increased after sprouting but underwent relative increase along digestion being the major phytochemicals found in the bioaccessible fraction obtained after intestinal digestion (73-94% of total phytochemicals). The increase in apigenin glycosides was associated to the increase of in vitro and intracellular antioxidant capacity of tabbouleh along digestion. Sprouting increased the peroxyl radical removal capacity of tabbouleh in the gastric phase and in the non-bioaccessible fraction regardless of the cultivar. The highest hydroxyl radical removal capacities were found in non-sprouted cultivars, especially in the soft texture cultivar in the undigested and bioaccessible fractions. The bioaccessible fraction obtained after wheat digestion was more efficient to scavenge intracellular ROS than undigested samples, the highest scavenging potency being observed for the hard texture cultivar with no effect of sprouting. These findings confirm the hypothesis that the phytochemicals of hard and soft wheat cultivars (sprouted or not) have different behavior during digestion in terms of biotransformation, bioaccessibility and ability to remove reactive species and indicate that tabbouleh produced from sprouted wheat results in increased release of bioactive phytochemicals during digestion.

Vitamin E reduces the reactive oxygen species production in dominant follicle during the negative energy balance in cattle.

In the postpartum period, there is an increase in non-esterified fatty acids (NEFA) in both serum and follicular fluid (FF) of cattle. The increase in fatty acid concentration results in increased production of reactive oxygen species (ROS) that can compromise bovine fertility. The objectives of this study were to characterize the lipid profile found in the FF of cows experiencing induced negative energy balance (NEB) and to evaluate the effect of α-tocopherol in the prevention of oxidative stress in the serum and FF of cows. Twenty-nine beef cows were divided into groups: (1) control; (2) Fasting for 24 days; and (3) Fasting + VitE. Between D0 and D4 blood samples were taken to assess concentrations of NEFA, ROS production, total antioxidant capacity (FRAP), lipid peroxidation, and α-tocopherol (vitamin E). On D4, follicular aspiration was performed for analysis of FF from the dominant follicle. Our results demonstrate that fasting was effective in causing increased fat mobilization in animals. The increase in serum concentration of C18:1c9 was reflected in the FF of fasting cows. Serum α-tocopherol concentration was higher in the control and Fasting + VitE groups compared to the Fasting group. In FF, there was an increase of α-tocopherol in the Fasting + VitE group in comparison to Fasting cows. There was an increase in ROS production in the serum of fasting cows. ROS production in FF was higher in the Fasting compared to the Fasting + VitE group. Vitamin E has beneficial effects in reducing ROS production in the dominant follicle of cows in NEB.

Effect of micronization on olive pomace biotransformation in the static model of colonic fermentation.

The effect of granulometric fractionation and micronization of olive pomace (OP) on the biotransformation of phenolic compounds by intestinal microbiota was investigated in vitro. Three types of powdered OP samples were incubated with human feces to simulate colonic fermentation, after a sequential static digestion: non-fractionated OP (NF), granulometrically fractionated OP (GF) and granulometrically fractionated and micronized OP (GFM). GF and GFM favored the release of hydroxytyrosol, oleuropein aglycone, apigenin and phenolic acid metabolites in the first hours of colonic fermentation compared to NF (up to 41-fold higher). GFM caused higher release of hydroxytyrosol than GF. GFM was the only sample to release tyrosol and sustained tyrosol levels up to 24 h of fermentation. Micronization associated with granulometric fractionation was more efficient than granulometric fractionation alone to increase the release of phenolic compounds from the OP matrix during simulated colonic fermentation and can be further studied for nutraceutical purposes.

Grain Germination Changes the Profile of Phenolic Compounds and Benzoxazinoids in Wheat: A Study on Hard and Soft Cultivars.

Pre-harvest sprouting is a frequent problem for wheat culture that can be simulated by laboratory-based germination. Despite reducing baking properties, wheat sprouting has been shown to increase the bioavailability of some nutrients. It was investigated whether wheat cultivars bearing distinct grain texture characteristics (BRS Guaraim, soft vs. BRS Marcante, hard texture) would have different behavior in terms of the changes in phytochemical compounds during germination. Using LC-Q-TOF-MS, higher contents of benzoxazinoids and flavonoids were found in the hard cultivar than in the soft one. Free phytochemicals, mainly benzoxazinoids, increased during germination in both cultivars. Before germination, soft and hard cultivars had a similar profile of matrix-bound phytochemicals, but during germination, these compounds have been shown to decrease only in the hard-texture cultivar, due to decreased levels of phenolic acids (trans-ferulic acid) and flavonoids (apigenin) that were bound to the cell wall through ester-type bonds. These findings confirm the hypothesis that hard and soft wheat cultivars have distinct behavior during germination concerning the changes in phytochemical compounds, namely the matrix-bound compounds. In addition, germination has been shown to remarkably increase the content of benzoxazinoids and the antioxidant capacity, which could bring a health-beneficial appeal for pre-harvested sprouted grains.

Effect of Adding Matricaria recutita L., Cymbopogon citratus, or Mentha piperita L. Extracts to Fermented Orange Beverage: Sensory Evaluation, Physicochemical Characterization, and Prediction of Toxic Risks and Biological Activity In Silico.

Fermentation is an important tool in producing functional beverages through agro-industrial wastes, and medicinal and aromatic plants due to the specific content of bioactive molecules. Therefore, this study evaluated the contribution of Matricaria recutita (chamomile), Cymbopogon citratus (lemongrass), or Mentha piperita (peppermint) extracts to the phytochemical profile and potential biological effects of a functional fermented orange beverage in vitro and in silico. The concentrations of aromatic herbal extracts that yielded the best sensory performance for fermented beverages were selected for analyses that involved characterizing the fermented beverages. The beverages that received the extracts (2%) had the highest phenolic and flavonoid content and antioxidant potential compared to the control. Hesperidin (124-130 mg L-1), narirutin (66-70 mg L-1), chlorogenic (11-16 mg L-1), caffeic (5.3-5.5 mg L-1), and ferulic (1-1.7 mg L-1) acids were found in the different formulations. The in silico analysis suggested that the evaluated compounds do not present a toxicity risk (mutagenicity, carcinogenicity, hepatotoxicity, and ability to penetrate the blood-brain barrier). Additionally, they can contribute to the biological effects of therapeutic importance, such as antioxidant, gastroprotective, and anti-ulcerative properties, and the Mentha piperita L. extract presented the greatest potential among the evaluated herbs for use in functional fermented beverages.

Collagen extracted from rabbit: Meat and by-products: Isolation and physicochemical assessment.

Obtaining collagen from rabbit meat, skin and ears is a great way to add value to these by-products. The collagen extracts from meat, skin, and ear showed high levels of protein 80.7, 95.5, and 94.5% and yields of 9.0, 24.4, and 23.8% on a dry basis, respectively. SDS-PAGE analysis showed that the collagens mainly consist of type I collagen, and the FTIR spectra displayed the characteristic peaks of amide A, B, I, II, and III; in addition, the collagens showed greater solubility in acidic pH. The foam production capacity of the collagens was low compared with other collagen sources. However, foam rabbit-collagen stability was high. The emulsifying activity index for the meat, skin, and ears was 44.7, 46.6, and 48.2 m2/g, respectively. Based on the results, the meat, skin, and ears of the rabbit proved to be a viable source for collagen extraction and a possible alternative to add value to the by-products (skin and ears) of these raw materials.

Norbixin, a natural dye that improves serum lipid profile in rabbits and prevents LDL oxidation.

We hypothesized that norbixin, which is a carotenoid used as an orange/red natural food coloring additive, has anti-atherogenic properties. An in vitro oxidation assay with human LDL and a rabbit model of atherosclerosis were used to test this hypothesis. Norbixin inhibited the oxidation of isolated human LDL in a concentration-dependent manner. In the in vivo assay, rabbits were fed with a regular chow (control) or an atherogenic diet (0.5% cholesterol) alone or supplemented with norbixin (10, 30 or 100 mg/kg b.w.) for 60 days. Norbixin supplementation (30 and 100 mg/kg b.w.) increased HDL levels and reduced triglyceride levels and the atherogenic index of rabbits. This effect was associated with the decrease of serum levels of oxidized LDL, oxidized LDL antibodies and aortic tissue levels of lipid and protein oxidation in the atherogenic rabbits supplemented with norbixin. Atherogenic diet increased enzymatic (superoxide dismutase, catalase, glutathione reductase, and thioredoxin reductase-1) and non-enzymatic (non-protein thiol groups content) antioxidant defense systems in the aortic tissue but reduced the activity of paraoxonase-1 in the serum. All these changes were prevented by norbixin supplementation (10, 30 and 100 mg/kg b.w.). These results suggest that norbixin has atheroprotective potential by improving serum lipid profile and preventing oxidative modifications of circulating LDL and aortic tissue. Norbixin may, therefore, be beneficial in the control of atherosclerosis risk factors and can be further investigated as a candidate to be used not only as a functional food ingredient but also for therapeutic applications and in the nutraceutical industry.

Acute effect of coffee on arterial stiffness and endothelial function in overweight and obese individuals: A randomized clinical trial.

INTRODUCTION: Coffee is one of the most consumed foodstuffs worldwide. Studies of coffee intake in healthy subjects have shown controversial effects on vascular function. However, little is known of coffee intake effects on the endothelium of overweight and obese individuals. OBJECTIVE: To investigate the acute effects of caffeinated and decaffeinated coffee intake on the endothelial function and arterial stiffness in overweight and obese individuals. METHODS: A randomized, double-blind, crossover clinical trial was designed to investigate the effects of regular caffeinated coffee and decaffeinated coffee on the endothelium. Each subject had both caffeinated coffee and decaffeinated coffee, separated by a washout period of seven days. The endothelial function was measured by flow-mediated dilation (FMD) assessed by ultrasound. Arterial stiffness was measured by an automatic oscillometric device. Blood samples were collected to assess the lipid and nitric oxide profiles. RESULTS: There were 18 subjects included in the study, aged 37.4 ± 10.0 years, with an average BMI of 28.96 ± 2.42, with the majority being female (61.1%). The caffeinated coffee increased central systolic blood pressure (P < 0.001), central diastolic blood pressure (P < 0.001) and pulse wave velocity (P < 0.001), but the decaffeinated coffee did not affect these variables. However, there was a better effect on FMD in the caffeinated coffee intake group (P = 0.014). CONCLUSION: In overweight and obese individuals, caffeinated coffee increased central blood pressure and pulse wave velocity but not the decaffeinated coffee. While caffeinated coffee showed an improvement on hyperemia-induced endothelial function. REGISTRATION NUMBER OF CLINICAL TRIAL: Platform of the Brazilian Registry of Clinical Trials under number RBR-65cxtr.

Grape UV-C irradiation in the postharvest period as a tool to improve sensorial quality and anthocyanin profile in 'Cabernet Sauvignon' wine.

Anthocyanins are important compounds in grapes and wine and significantly influence their characteristics. Ultraviolet light-C (UV-C) can be used as a tool to induce secondary metabolism, and in this study, it was used in 'Cabernet Sauvignon' grapes in the postharvest period to improve the anthocyanin profile and sensory attributes in wine produced with irradiated grapes. Grapes in the postharvest period were exposed to 0, 2, or 3 kJ m-2 UV-C radiation. After winemaking and storage time (6 months), physicochemical and color analyses, anthocyanin quantification and identification (HPLC-PDA-MS/MS), and sensory analyses were carried out. 'Cabernet Sauvignon' wine produced with grapes irradiated with 3 kJ m-2 UV-C showed increased spectrophotometric color, which is likely due to a 22.5% increase in total anthocyanin monomers, 59.3% of pyranoanthocyanins, 92.3% of direct condensation products, and 62.8% of acetaldehyde-mediated condensation products. In addition, this irradiated dose presented higher perception scores for visual color, aroma, taste, and was preferred by the tasters over the wine produced with non-irradiated grapes. This study is the first of its kind to show that the UV-C radiation treatment of grapes in the postharvest period can be a promising tool to improve the anthocyanin profile and sensorial quality of 'Cabernet Sauvignon' wine. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05191-5.

Strategy to increase the lipid stability of the microbial oil produced by Umbelopsis isabellina for food purposes: Use of microencapsulation by external ionic gelation.

Oleaginous microorganisms, including the fungus Umbelopsis isabellina, have emerged as a biotechnological alternative to obtain polyunsaturated fatty acid-rich oils, which are strongly linked to energy purposes (biofuel) than the food industry. Considering the composition of microbial oil and its use by the food industry, it is necessary to investigate strategies that increase its lipid stability. Ergo, this pioneering study aimed to microencapsulate the oil produced by Umbelopsis isabellina and evaluate its oxidative stability throughout the storage period against factors such as temperature and luminosity. The microbial oil was microencapsulated through the external ionic gelation technique, producing an encapsulation efficiency of 80% and proving to be a suitable method because it maintained oil composition. Combining microencapsulation and refrigerated storage led to the best effects on storage time, increasing the evaluated lipid stability through the peroxide values and conjugated diene formation. Moreover, saturated and monounsaturated fatty acid content increased, and polyunsaturated fatty acid content decreased during storage for both the free and microencapsulated oil, regardless of storage temperature, although microencapsulation reduced the changes. The results primarily demonstrate how microencapsulation prolongs the oxidative stability and unsaturated fatty acid content of the microbial oil by reducing its reactions to external environmental factors, thus facilitating its use in the food industry.

Antimycobacterial activity of Achyrocline flaccida (Asteraceae) aqueous extract from Southern Brazil.

Achyrocline flaccida aqueous extract was obtained by macerating wildflowers. The phytochemical profile present in the A. flaccida aqueous extract was elucidated by HPLC-ESI-MS/MS. Toxicity was evaluated in vitro by comet assay in peripheral blood mononuclear cells (PBMCs) and in vivo using Caenorhabditis elegans as a model. The antioxidant activity was also evaluated, and antimycobacterial activity was assessed by the broth microdilution method. The compounds present in the aqueous extract mainly belonged to the flavonoid class (89%). The concentrations that showed protective effects in C. elegans against oxidative stress and antimycobacterial activity had no toxic effects. The antimycobacterial activity test demonstrated that the concentration of 1,560 µg mL-1 inhibited the growth and eradication of the mycobacterial tested strains. Based on our findings, the A. flaccida aqueous extract presents a viable potential in developing new phytotherapeutic drugs against mycobacteria of clinical relevance.

Tucumã (Astrocaryum aculeatum) extract: phytochemical characterization, acute and subacute oral toxicity studies in Wistar rats.

Natural products are often used by the population to treat and/or prevent several disorders. Tucumã is an Amazonian fruit widely consumed by local population and no in vivo toxicity studies regarding its safety are available in the literature to date. Therefore, the phytochemical characterization, acute and repeated dose 28-day oral toxicities of crude extract of tucumã's pulp (CETP) in Wistar rats were evaluated. For the CETP preparation, tucumã pulp was crushed and placed into sealed amber glass jars containing absolute ethanol solution for extraction. CETP phytochemical analyses evidenced the presence of carotenoids, flavonoids, unsaturated and satured fatty acids, and triterpenes. In the acute toxicity, female rats from the test group were treated with CETP at single dose of 2000 mg/kg. For the repeated dose toxicity, CETP was administered to male and female rats at doses of 200, 400 and 600 mg/kg, for 28 days. Body weight was recorded during the experiment and blood, liver and kidney were collected for further analysis. No mortality or toxicity signs were observed during the studies. CETP was classified as safe (category 5, OECD guide), in acute toxicity. In repeated dose study was observed alterations in some biochemical parameters, as well as in oxidative damage and enzymatic activity. Histopathological findings showed renal damage in male rats at higher dose. The data obtained suggest that CETP did not induced toxicity after exposure to a single or repeated doses in female rats. However, in males may be considered safe when given repeatedly in low doses.

Enhancement of the Functional Properties of Mead Aged with Oak (Quercus) Chips at Different Toasting Levels.

Consumers increasingly prefer and seek functional beverages, which, given their characteristics, provide important bioactive compounds that help prevent and treat chronic diseases. Mead is a traditional fermented alcoholic beverage made from honey solution. The aging process of mead with oak chips is innovative and bestows functional characteristics to this beverage. Thus, in this study, we sought to develop and characterize a novel functional beverage by combining the health benefits of honey with the traditional aging process of alcoholic beverages in wood. Phenolic compounds, flavonoids, and antioxidant capacity were analyzed in mead using oak chips at different toasting levels and aged for 360 days. LC-ESI-QTOF-MS/MS was used to analyze the chemical profile of different meads. Over time, the aging process with oak chips showed a higher total phenolic and flavonoid content and antioxidant capacity. Eighteen compounds belonging to the classes of organic acids, phenolic acids, flavonoids, and tannins were identified in meads after 360 days. Our findings revealed that the addition of oak chips during aging contributed to p-coumaric, ellagic, abscisic, and chlorogenic acids, and naringenin, vanillin, and tiliroside significantly impacted the functional quality of mead.

Natural deep eutectic solvent (NADES): A strategy to improve the bioavailability of blueberry phenolic compounds in a ready-to-use extract.

This study investigated whether a ready-to-use extract obtained using a natural deep eutectic solvent (NADES) affects the pharmacokinetic profile of blueberry phenolic compounds compared to organic solvent (SORG)-extracted compounds. SORG extract was administered as an aqueous solution after solvent removal. Wistar rats received a single dose of crude extract of blueberry obtained using NADES (CE-NADES) or SORG (CE-SORG), followed by LC-DAD-MS/MS analysis of blood and cecal feces. Non-compartmental pharmacokinetic analysis revealed that CE-NADES increased the bioavailability of anthocyanins by 140% compared to CE-SORG. CE-NADES increased the stability of phenolic compounds during in vitro digestion by delaying gastric chyme neutralization. These results suggest that besides being an eco-friendly solvent for the extraction of phytochemicals, choline chloride:glycerol:citric acid-based NADES can be used as a ready-to-use vehicle for increasing oral absorption of bioactive compounds such as anthocyanins.

Goji berry (Lycium barbarum L.) juice reduces lifespan and premature aging of Caenorhabditis elegans: Is it safe to consume it?

Goji berry fruit is considered a healthy food. However, studies on its effects on aging and safety are rare. This study is the first to evaluate the effects of goji berry juice (GBJ) on oxidative stress, metabolic markers, and lifespan of Caenorhabditis elegans. GBJ caused toxicity, reduced the lifespan of C. elegans by 50%, and increased the reactive oxygen species (ROS) production by 45-50% at all tested concentrations (1-20 mg/µL) of GBJ. Moreover, the highest concentration of GBJ increased lipid peroxidation by 80% and altered the antioxidant enzymes. These effects could be attributed to a pro-oxidant effect induced by GBJ polyphenols and carotenoids. Moreover, GBJ increased lipofuscin, glucose levels, number of apoptotic bodies, and lipase activity. The use of mutant strains demonstrated that these effects observed in the worms treated with GBJ were not associated with the Daf-16/FOXO or SKN-1 pathways. Our findings revealed that GBJ (mainly the highest concentration) exerted toxic effects and promoted premature aging in C. elegans. Therefore, its consumption should be carefully considered until further studies in mammals are conducted.

Achyrocline flaccida essential oil from Brazil: phytochemical composition, genotoxicity, protective effects on Caenorhabditis elegans, and antimycobacterial activity.

Achyrocline flaccida (Weinm) DC. essential oil was obtained by hydrodistillation of wild flowers from southern Brazil. We explored, for the first time, the phytochemical composition, toxicity, resistance to oxidative stress in Caenorhabditis elegans, and antimycobacterial activities of A. flaccida essential oil. Twenty-four compounds were identified by GC-FID-MS and the major constituents identified were α-pinene (41.10%) and caryophyllene (30.52%). The essential oil showed no signs of genotoxicity in the comet assay and presented relevant antioxidant capacity since it prevented ROS production in the C. elegans model. Furthermore, the minimal inhibitory concentrations (MIC) results showed that M. abscessus, M. massiliense, and M. fortuitum had their growth inhibited by A. flaccida essential oil. Therefore, the essential oil of this plant is a promising alternative in the search for new compounds capable of decreasing oxidative stress and treating mycobacteriosis.

Natural deep eutectic solvent (NADES)-based blueberry extracts protect against ethanol-induced gastric ulcer in rats.

Blueberry is a polyphenol-rich fruit bearing great bioactive potential. Natural deep eutectic solvents (NADES) emerged as putatively biocompatible solvents that could substitute for toxic organic solvents in the extraction of fruit phenolic compounds for developing nutraceuticals or functional foods. Therefore, the aim of this study was to investigate the gastroprotective effects and the biocompatibility of a blueberry crude extract (CE) obtained using NADES and of the extract fractions (anthocyanin-rich fraction - ARF; non-anthocyanin phenolic fraction - NAPF) in a model of ethanol-induced gastric ulcer in rats. CE was the NADES-containing, ready-to-use extract that was obtained using choline chloride:glycerol:citric acid NADES (0.5:2:0.5 M ratio). ARF and NAPF were the NADES-free fractions obtained by solid phase purification of CE and were investigated to identify the bioactive fraction responsible for the effects of CE. Animals were treated for 14 days with water, NADES vehicle, CE, ARF, NAPF or lansoprazole (intragastric) and then received ethanol to induce gastric ulcer. CE decreased ulcer index and preserved the integrity of gastric mucosa. The pretreatment with CE or ARF reduced glutathione depletion and the inflammatory response. All treatments, including NADES vehicle reduced protein oxidation and nitric oxide overproduction in ethanol-treated rats. Additionally, ARF increased short-chain fatty acids in feces. These findings suggest that NADES can be used to obtain biocompatible extracts of blueberry that exhibit gastroprotective effects with no need of solvent removal. The gastroprotective effects were mainly associated to ARF but NAPF and even NADES vehicle also contributed to some protective effects.

Bioactive compounds, antioxidant capacity and antitumoral activity of ethanolic extracts from fruits and seeds of Eugenia involucrata DC.

Eugenia involucrata DC. (Myrtaceae) is a native tree species from Brazil that has been scarcely studied. We investigated the phenolic composition, the antioxidant capacity and the antitumoral activity of ethanolic extracts from fruits (FE) and seeds (SE) of E. involucrata. Six anthocyanins were identified by UPLC-PDA/MS/MS in FE, being four derived from cyanidin, and the other ones derived from delphinidin and pelargonidin. Using HPLC-PDA, FE presented a larger number of phenolic compounds (epicatechin, catechin, rutin, ellagic acid, myricetin and quercetin) than SE, which did not show myricetin and quercetin. However, SE showed higher total phenolic content and generally stronger in vitro antioxidant capacity than FE, except that only FE exhibited superoxide radical scavenging activity, which may be attributed to the anthocyanins present in fruits. Additionally, only SE exhibited antitumoral activity in a pancreatic cancer cell line (PANC-1). The antitumoral mechanisms involved imbalance of antioxidant status, alteration of mitochondrial membrane potential, cytoskeleton disassembly and induction of cell death by apoptosis and necrosis. Compared to the standard antitumoral drug gemcitabine, SE exhibited higher antitumoral efficacy and selectivity index. The highest concentration of total phenolics and of specific phenolic compounds bearing antitumoral properties may be related to the antitumoral activity of SE. Our results corroborate previous data of E. involucrata as an important source of bioactive compounds and provide, for the first time, evidences of in vitro antitumoral potential of its seeds on pancreatic cancer cell line.