Effects of simulated gastrointestinal digestion/epithelial transport on phenolics and bioactivities of particles of brewer's spent yeasts loaded with Brazilian red propolis.

Red propolis from northeast Brazil contains mainly isoflavonoids as bioactive compounds, and its consumption may counteract unregulated and exacerbated formation of reactive oxygen species and inflammatory cytokines/chemokines. Moreover, the production of particles using sustainable carriers have been studied to increase the use of propolis as a functional food ingredient. Hence, the objective of this work was to investigate the effects of simulated gastrointestinal digestion followed by a cell-based epithelial transport on phenolic profile, anti-inflammatory and antioxidant activities of particles of brewer's spent yeasts (BSY) loaded with ethanolic extract of Brazilian red propolis (EEP). As a result, the EEP phenolic diversity decreased throughout the simulated gastrointestinal system, and was modulated by the particle production, as detected by high-performance liquid chromatography - electrospray ionization - quadrupole-time-of-flight-mass spectrometry (HPLC-ESI-QTOF-MS). Concomitantly, the antioxidant activity, as assessed by the ability to scavenge peroxyl and superoxide radicals, hydrogen peroxide, and hypochlorous acid, generally decreased at a higher extent for the particles of EEP with BSY (EEP-BSY) throughout the experiments. Nonetheless, after epithelial transport through the Caco-2 cell monolayer, the basolateral fraction of both EEP-BSY and EEP decreased the activation of pro-inflammatory transcription factor NF-κB by 83% and 65%, respectively, as well as the release of TNF-α (up to 51% and 38%, respectively), and CXCL2/MIP-2 (up to 33% and 25%, respectively). Therefore, BSY may be an interesting carrier for EEP bioencapsulation, since it preserves its anti-inflammatory activity. Further studies should be encouraged to investigate the feasibility of adding it in formulations of functional foods, considering its effect on sensory attributes.

Bioactive Compounds and In Vitro Antioxidant Capacity of Cambuci and Uvaia: An Extensive Description of Little-Known Fruits from the Myrtaceae Family with High Consumption Potential.

Cambuci (Campomanesia phaea O. Berg Landrum) and uvaia (Eugenia pyriformis Cambess), both native Atlantic Rainforest fruits, are noteworthy for being rich in bioactive compounds and their significant antioxidant capacity. Despite the numerous known edible fruits in the world, consumption by humans is most often restricted to a few dozen of them. Such behavior occurs, among other reasons, due to the lack of knowledge about fruits not yet commercialized on a large scale. This study quantified the bioactive compound content (total phenolic compounds and ascorbic acid in cambucis and uvaias; proanthocyanidins in cambucis, and total carotenoid profile and individual carotenoids for grapes) and antioxidant capacity of the edible parts (peel and pulp) of cambuci and uvaia accessions, using three methods (ABTS•+, ROO• radical scavenging and HOCl elimination). Cambuci contained higher phenolic compound levels and displayed higher antioxidant capacity determined by the ABTS•+ and ROO• radical scavenging methods than uvaia (139 and 119 mg 100 g-1 of GAE, 10.5 and 7.73 µmol g-1 of TE; 9.17 and 5.92 µmol g-1 of TE, respectively). Vitamin C content and the antioxidant capacity determined by the HOCl elimination method were about 1.5- and 6-fold higher in uvaia compared to cambuci, with the latter being a first-time report for uvaia. Both fruits contained higher levels of bioactive compounds and antioxidant capacity than other commonly consumed fruits.

The phytoactive constituents of Eugenia selloi B.D. Jacks (pitangatuba): Toxicity and elucidation of their anti-inflammatory mechanism(s) of action.

We determined the phytochemical composition, anti-inflammatory mechanism of action, ROS/RNS scavenging capacity and systemic toxicity of a purified subfraction (S8) of Eugenia selloi. The composition of S8 was assessed by LC-ESI-QTOF-MS; the anti-inflammatory activity in RAW264.7 macrophages through NF-κB activation and biomarkers by multiplex in THP-1 cells; neutrophil migration, intravital microscopy and ICAM-1 expression in mice; NETs formation and CD11b expression; S8 scavenging capacity of ROS/RNS; toxicity in Galleria mellonella larvae model. Coumaric acid, quercetrin and vanillic acid were identified. S8 decreased NF-κB activation, IL-1ß, IL-6, IL-10, MDC and MCP-1 levels, reduced neutrophil migration and ICAM-1 expression in mice; S8 did not interfere NET formation and CD11b expression, exhibited high antioxidant and showed negligible toxicity. E. selloi proved to be a promising, yet underexplored source of bioactive compounds, which can be useful employed in agribusiness and in the pharmaceutical and food industry to develop new products or human health supplies.

Lignans as new chemical markers of a certified Brazilian organic propolis.

Commercially certified organic propolis produced in areas of environmental conservation and reforestation forests of Southern Brazil are generally poor in flavonoids, although one of its variants - Brazilian certified organic propolis 1 (OP1) - has shown strong antioxidant activity. The objective was to identify active compounds from OP1 related to its strong antioxidant activity. OP1 ethanolic extracts were subjected to liquid-liquid fractionation, and the fractions presenting the strongest antioxidant activity were combined and purified into subfractions. Compounds isolated from the most active subfractions had their structure elucidated by Nuclear Magnetic Resonance (NMR). As a result, five lignans and two lignan-precursors were isolated, and four of them are herein reported for the very first time in propolis. Hence, these compounds may be used as chemical markers for product standardization and authentication purposes, since OP1 is only produced by honeybees in native forests and its botanical origins remain unknown.

The phytoactive constituents of Eugenia selloi B.D. Jacks (pitangatuba): toxicity and elucidation of their anti-inflammatory mechanism(s) of action

We determined the phytochemical composition, anti-inflammatory mechanism of action, ROS/RNS scavenging capacity and systemic toxicity of a purified subfraction (S8) of Eugenia selloi. The composition of S8 was assessed by LC-ESI-QTOF-MS; the anti-inflammatory activity in RAW264.7 macrophages through NF-κB activation and biomarkers by multiplex in THP-1 cells; neutrophil migration, intravital microscopy and ICAM-1 expression in mice; NETs formation and CD11b expression; S8 scavenging capacity of ROS/RNS; toxicity in Galleria mellonella larvae model. Coumaric acid, quercetrin and vanillic acid were identified. S8 decreased NF-κB activation, IL-1β, IL-6, IL-10, MDC and MCP-1 levels, reduced neutrophil migration and ICAM-1 expression in mice; S8 did not interfere NET formation and CD11b expression, exhibited high antioxidant and showed negligible toxicity. E. selloi proved to be a promising, yet underexplored source of bioactive compounds, which can be useful employed in agribusiness and in the pharmaceutical and food industry to develop new products or human health supplies.

Active Antioxidant Phenolics from Brazilian Red Propolis: An Optimization Study for Their Recovery and Identification by LC-ESI-QTOF-MS/MS.

Brazilian red propolis (BRP) is a natural product widely known for its phenolic composition and strong antioxidant properties. In this study, we used the Box-Behnken Design (BBD) with Surface Response Methodology to optimize the extraction conditions for total phenolic content (TPC) and Trolox equivalent antioxidant capacity(TEAC) of bioactive phenolics from BRP. The extraction time, ethanol/water concentration and temperature, were tested. All variables had significant effects (p ≤ 0.05), with a desirability coefficient of 0.88. Under optimized conditions (90% ethanol at 80 °C for 30 min), the BRP extract showed a TPC of 129.00 ± 2.16 mg GAE/g and a TEAC of 3471.76 ± 53.86 µmol TE/g. Moreover, FRAP and ORAC assays revealed that the optimized BRP extract had 1472.86 ± 72.37 µmol Fe2+/g and 4339.61 ± 114.65 µmol TE/gof dry weight, respectively. Thirty-two phenolic compounds were tentatively identified by LC-QTOF-ESI-MS/MS, of which thirteen were found for the first time in BRP, including four flavones, one flavanol, two flavanones, two chalcones, and four isoflavonoids. Thus, our results highlight the importance of BRP as a source of a wide variety of phenolic compounds with significant antioxidant properties.

Antihyperglycemic activity of crude extract and isolation of phenolic compounds with antioxidant activity from Moringa oleifera Lam. leaves grown in Southern Brazil.

The antihyperglicemic activity of crude extract from Moringa oleifera leaves and isolation of phenolic compounds with antioxidant activity using bioguided assay were employed by the first time in leaves cultivated in Brazil. The hydroalcoholic extract (HE) was produced by using ethanol:water (80:20 v/v) and purified by solid-liquid procedure using solvents in ascending order of polarity. The ethyl acetate fraction (Fr-EtOAc) presented high antioxidant potential and it was purified using chromatographic techniques rendering isolated compounds that were identified from the spectral data. The HE extract (500 mg kg-1) was adimistrated in diabetic rats induced by streptozotocin and chemical markers and lipid peroxidation in liver and kidney were evaluated. The Fr-EtOAc showed high antioxidant potential by FRAP reduction method (1678 µmol Fe2+ g-1), DPPH and ABTS scavenging methods (526.7 and 671.5 µmol TEAC g-1 respectively) and ORAC assay (3560.6 µmol TEAC g-1). Therefore, the Fr-EtOAc was purified and yielded three bioactive subfractions (S-12, S-13 abd S-15) that were rechromatoghaphed in HPLC-SemiPrep. After that, two main bioactive glycosylated flavonoids (isoquercitrin and astragalin) and phenolic acid (3-O-caffeoylquinic acid) were obtained. Additionally, the HE extract provided protection against oxidative damage in liver and kidney of diabetic rats ameliorating endogenous antioxidant defenses by increase catalase (CAT), glutathione S-transferase (GST) and non-protein thiol groups (NPSH) levels as well as decreased the lipid peroxidation in these tissues. Our results indicate that three phenolic compounds with high antioxidant activity were isolated and, the chemical composition of HE crude extract, rich in flavonoids glycosylated could be intimately related to antihyperglycemic action. So, it is possible to suggest that these compounds may be used as chemical biomarkers for this plant in Brazil, ensuring quality and supporting the use of aerial parts in tradicional medicine.

Simulated gastrointestinal digestion of Brazilian açaí seeds affects the content of flavan-3-ol derivatives, and their antioxidant and anti-inflammatory activities.

Açaí seeds (Euterpe oleracea Mart.) are the major residue generated during industrial extraction of açaí fruit pulp - a popular and typical Amazon fruit rich in bioactive compounds and nutrients. In this study, we investigated the bioaccessibility of an açaí seed extract using an in vitro simulated gastrointestinal digestion model. Catechin, epicatechin and procyanidins B1 and B2 were identified and quantified in the açaí seed extract and monitored by HPLC-DAD through the digestion phases. Bioaccessibility of these flavan-3-ols and deactivation of reactive oxygen species decreased after the intestinal phase, except for peroxyl radical (ROO●). RAW 264.7 macrophages treated either with the digested or undigested açaí seed extract showed reduced NF-κB activation and TNF-α levels, even following gastrointestinal digestion. Thus, the ROO● scavenging capacity and anti-inflammatory activity of the extract were found to be still remarkable after digestion, suggesting that açaí seeds could be explored as a source of bioactive compounds for functional foods, cosmetic or pharmaceutical purposes.

Phenolic profile and potential beneficial effects of underutilized Brazilian native fruits on scavenging of ROS and RNS and anti-inflammatory and antimicrobial properties.

Brazilian native fruits (BNF) have aroused interest of researchers and consumers for their great human health benefits. In this study, five BNF (Byrsonima lancifolia, Campomanesia phaea, Jacaratia spinosa, Solanum alternatopinnatum and Acnistus arborescens) were tested for their polyphenolic compounds by LC-ESI-MS/MS, reactive species deactivation (ROO˙, O2˙-, HOCl and NO˙), anti-inflammatory properties in vivo, and in vitro antimicrobial activity - with determination of putative mechanism(s) of action. Eighty-one polyphenols were identified, which exhibited a significant capacity to deactivate both ROS and RNS. C. phaea extract had the highest capacity to scavenge ROO˙ (68.94 µmol TE per g), O2˙- (IC50: 575.36 µg mL-1) and NO˙ (IC50: 16.96 µg mL-1), which may be attributed to the presence of ellagitanins. B. lancifolia decreased neutrophil influx into the peritoneal cavity of mice by 50% as compared to carrageenan and reduced Candida albicans biofilm viability by 3 log10 possibly due to complexation with cell membrane ergosterol. In summary, the BNF presented herein are good sources of bioactive compounds with positive effects on deactivation of biological reactive species, as well as with anti-inflammatory and antimicrobial activities, which can be altogether highly beneficial to human health.

Anti-inflammatory and antioxidant potential, in vivo toxicity, and polyphenolic composition of Eugenia selloi B.D.Jacks. (pitangatuba), a Brazilian native fruit.

Brazilian native fruits are a rich source of polyphenolic compounds that can act as anti-inflammatory and antioxidant agents. Here, we determined the polyphenolic composition, anti-inflammatory mechanism of action, antioxidant activity and systemic toxicity in Galleria mellonella larvae of Eugenia selloi B.D.Jacks. (synonym Eugenia neonitida Sobral) extract (Ese) and its polyphenol-rich fraction (F3) obtained through bioassay-guided fractionation. Phenolic compounds present in Ese and F3 were identified by LC-ESI-QTOF-MS. The anti-inflammatory activity of Ese and F3 was tested in vitro and in vivo through NF-κB activation, cytokine release and neutrophil migration assays. The samples were tested for their effects against reactive species (ROO•, O2•-, HOCl and NO•) and for their toxicity in Galleria mellonella larvae model. The presence of hydroxybenzoic acid, ellagitannins and flavonoids was identified. Ese and F3 reduced NF-κB activation, cytokine release and neutrophil migration, with F3 being three-fold more potent. Overall, F3 exhibited strong antioxidant effects against biologically relevant radicals, and neither Ese nor F3 were toxic to G. mellonella larvae. In conclusion, Ese and F3 revealed the presence of phenolic compounds that decreased the inflammatory parameters evaluated and inhibited reactive oxygen/nitrogen species. E. selloi is a novel source of bioactive compounds that may provide benefits for human health.

Comprehensive characterization of bioactive phenols from new Brazilian superfruits by LC-ESI-QTOF-MS, and their ROS and RNS scavenging effects and anti-inflammatory activity.

Brazilian native fruits (BNF) remain unexplored and underutilized resources with a high potential to improve human health and wellness. In our study, five new BNF (Eugenia stipitata, Sageretia elegans, Byrsonima arthropoda, Spondias mombin andRubus rosaefolius)were evaluated for their phenolic composition by LC-ESI-QTOF-MS; and for their ROS and RNS scavenging effects (ROO, O2-, NO, HOCl); in vivo anti-inflammatory activity (neutrophil migration); and in vivo acute toxicity in Galleria mellonella. Eighty-six phenolic compounds were identified, including hydroxybenzoic acids, hydroxycinnamic acids, flavonoids, anthocyanins and ellagitannins, several of which had never been reported in BNF. The BNF exhibited high antioxidant effects against biologically relevant radicals, and treated animals showed decreased neutrophil influx and NF-kB activation. Thus, these BNF are good sources of antioxidant and anti-inflammatory molecules that can be beneficial for human health as functional foods. Based on their bioactivity, they can be considered as new Brazilian superfruits.

Exploration of avocado by-products as natural sources of bioactive compounds.

This study aimed to evaluate the antioxidant, anti-inflammatory, and cytotoxic properties and phenolic composition of peel and seed of avocado varieties Hass and Fuerte using green solvents. Ethanol soluble compounds were identified in peel and seed of both varieties using HPLC-MS/MS and quantified using HPLC-DAD. Agro-industrial by-products of both varieties exhibited high radical scavenging activity against synthetic free radicals (DPPH and ABTS) and reactive oxygen species (peroxyl, superoxide, and hypochlorous acid) and high ability to reduce Fe3+ to Fe2+. The main compounds with significant contribution to the antioxidant activity determined by online HPLC-ABTS●+ analyses were procyanidin B2 and epicatechin in the peel and trans-5-O-caffeoyl-D-quinic acid, procyanidin B1, catechin, and epicatechin in the seed. Peel of Fuerte significantly suppressed TNF-α and nitric oxide (NO) release (459.3 pg/mL and 8.5 µM, respectively), possibly because of the high phenolic content and antioxidant activity detected. Avocado agro-industrial by-products can be used for food and pharmaceutical purposes due to their antioxidant and anti-inflammatory properties.

Volatile and non-volatile/semi-volatile compounds and in vitro bioactive properties of Chilean Ulmo (Eucryphia cordifolia Cav.) honey.

Ulmo honey originating from Eucryphia cordifolia tree, known locally in the Araucania region as the Ulmo tree is a natural product with valuable nutritional and medicinal qualities. It has been used in the Mapuche culture to treat infections. This study aimed to identify the volatile and non-volatile/semi-volatile compounds of Ulmo honey and elucidate its in vitro biological properties by evaluating its antioxidant, antibacterial, antiproliferative and hemolytic properties and cytotoxicity in Caco-2 cells. Headspace volatiles of Ulmo honey were isolated by solid-phase microextraction (SPME); non-volatiles/semi-volatiles were obtained by removing all saccharides with acidified water and the compounds were identified by GC/MS analysis. Ulmo honey volatiles consisted of 50 compounds predominated by 20 flavor components. Two of the volatile compounds, lyrame and anethol have never been reported before as honey compounds. The non-volatile/semi-volatile components of Ulmo honey comprised 27 compounds including 13 benzene derivatives accounting 75% of the total peak area. Ulmo honey exhibited weak antioxidant activity but strong antibacterial activity particularly against gram-negative bacteria and methicillin-resistant Staphylococcus aureus (MRSA), the main strain involved in wounds and skin infections. At concentrations >0.5%, Ulmo honey reduced Caco-2 cell viability, released lactate dehydrogenase (LDH) and increased reactive oxygen species (ROS) production in a dose dependent manner in the presence of foetal bovine serum (FBS). The wide array of volatile and non-volatile/semi-volatile constituents of Ulmo honey rich in benzene derivatives may partly account for its strong antibacterial and antiproliferative properties important for its therapeutic use. Our results indicate that Ulmo honey can potentially inhibit cancer growth at least partly by modulating oxidative stress.

Anti-Inflammatory, Anti-Osteoclastogenic and Antioxidant Effects of Malva sylvestris Extract and Fractions: In Vitro and In Vivo Studies.

Given their medical importance, natural products represent a tremendous source of drug discovery. The aim of this study was to investigate Malva sylvestris L. extract and fractions and their pharmacological activities followed by chemical identification. The aqueous fraction (AF) was identified as the bioactive fraction in the in vitro and in vivo assays. The AF controlled the neutrophil migration to the peritoneal cavity by 66%, inhibited the antiedematogenic activity by 58.8%, and controlled IL-1ß cytokine expression by 54%. The in vitro viability tests showed a concentration-dependent effect, where the MSE and fractions at concentrations under 10 µg/mL were non-toxic to cells. Transcriptional factors of carbonic anhydrase II (CAII), cathepsin K (Ctsk) and tartrate-resistant acid phosphatase (TRAP) were analyzed by qPCR in RAW 264.7 cell lines. The gene expression analysis showed that the AF was the only treatment that could downregulate all the study genes: CAII, Ctsk and TRAP (p<0.05). TRAP staining was used to evaluate osteoclast formation. AF treatments reduced the number of osteoclastogenesis 2.6-fold compared to the vehicle control group. Matrix metalloproteinase 9 (MMP-9) activity decreased 75% with the AF treatment. Moreover, the bioactive fraction had the ability to regulate the oxidation pathway in the ABTS (2,2-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) assay with an activity equivalent to 1.30 µmol Trolox/g and DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals 1.01 g/L. Positive ion ESI-mass spectrometry for molecular ions at m/z 611 and 633 confirmed rutin as the major compound in the AF. The AF of M. sylvestris presented anti-inflammatory, controlled osteoclastogenic mechanisms and antioxidant abilities in different in vitro and in vivo methods. In addition, we suggest that given its multi-target activity the bioactive fraction may be a good candidate in the therapy of chronic inflammatory diseases.

Extraction yield, antioxidant activity andphenolics from grape, mango and peanut agro-industrial by-products / Rendimento de extração, atividade antioxidante e compostos fenólicos dos subprodutos agro industriais de uva, manga e amendoim

ABSTRACT: The objective of this study was to determine and correlate the extraction yields, antioxidant activity, total phenolics and total flavonoids from grape, mango and peanut agro-industrial by-products. The β-carotene/linoleic acid autoxidation system and scavenging capacity for DPPH and ABTS free radicals assays were used. The results were expressed in terms of lyophilized sample or dry extract. Mango bagasse exhibited the highest extraction yield (37.07%) followed by peanut skin (15.17%) and grape marc (7.92%). In terms of lyophilized sample, total phenolics did not vary significantly among the residues evaluated (average of 60.33mg EAG g-1); however, when they were expressed as dry extract grape marc exhibited the highest total phenolic (768.56±116.35mg GAE g-1), followed by peanut skin (404.40±13.22mg GAE g-1) and mango bagasse (160.25±4.52mg GAE g-1), Peanut skin exhibited the highest content of total flavonoids (2.44mg QE g-1), while grape marc (1.76mg QE g-1) and mango bagasse (1.70 mg QE g-1) showed no significant differences. The extraction yield showed strong negative linear correlation with total phenolic and total flavonoid. This study showed that peanut skin was the sample with the highest antioxidant activity and it was strongly influenced by total flavonoids. All extracts of byproducts showed antioxidant activity comparable to α-tocopherol, and they can be a source of natural compounds with potential to replace synthetic antioxidants such as BHT.

RESUMO: O objetivo deste estudo foi determinar e correlacionar os rendimentos de extração, atividade antioxidante, compostos fenólicos totais e flavonoides de subprodutos agroindustriais de uva, manga e amendoim. A autoxidação do sistema β-caroteno/ácido linoleico e a capacidade de sequestro dos radicais DPPH e ABTS foram os ensaios antioxidantes utilizados. Os resultados foram expressos com base na amostra liofilizada ou no extrato seco. O bagaço de manga apresentou o maior rendimento de extração (37,07%) seguido da película de amendoim (15,17%) e bagaço de uva (7,92%). Em termos de amostra liofilizada, o teor de fenólicos totais não variou significativamente entre os resíduos avaliados (média de 60,33mg EAG g-1), entretanto, quando foi expresso com base no extrato seco, o bagaço de uva exibiu o maior teor de fenólicos totais (768,56±116,35mg EAG g-1), seguido da película de amendoim (404,40±13,22mg EAG g-1) e bagaço de manga (160,25±4,52 mg EAG g-1). Apelícula de amendoim apresentou o maior conteúdo de flavonoides totais (2,44mg EQ g-1), enquanto os bagaços de uva (1,76mg EQ g-1) e de manga (1,70mg EQ g-1) não mostraram diferenças significativas. O rendimento de extração mostrou fortes correlações lineares negativas como teor de fenólicos totais e flavonoides totais. Este estudo mostrou que a películade amendoim foi o material que apresentou a maior atividade antioxidante e isso foi fortemente influenciado por flavonoides totais. Os extratos avaliados mostraram atividade antioxidante comparável à do α-tocoferol, com potencial para substituir antioxidantes sintéticos, tais como o BHT.

Physicochemical, Functional and Antioxidant Properties of Tropical Fruits Co-products.

The aim of this study was to determine the physicochemical, functional and antioxidant properties of mango (MAC), pineapple (PAC) and passion fruit (PFC) co-products in order to evaluate them as ingredients for food application. Proximate composition showed low fat content (0.95-5.64 g/100 g), and high levels of dietary fiber. In pineapple and passion fruit co-products, dietary fiber represented more than 50 % of the sample. Low pH, water activity, along with high acidity indicated that these co-products would not be easily susceptible to deterioration as food ingredients. Pineapple and passion fruit co-products had significant (p < 0.05) water holding capacity (4.96 and 4.31 g water/g sample, respectively), however oil holding capacity was low (1.59-1.85 g oil/g sample) for the three matrices studied. Regarding the phenolic content, values ranged from 3.78 to 4.67 mg gallic acid equivalent/g, with MAC showing the highest content. Through high performance liquid chromatography analysis, six compounds were identified and quantified (gallic acid, p-coumaric acid, ferulic acid, caffeic acid, epicatechin, and mangiferin) in the fruit co-products. As observed for the phenolic content, the highest antioxidant activity (p < 0.05) was found in MAC when measured by both DPPH and ABTS methods. The results indicated that the fruit co-products under evaluation could be used as functional ingredient to provide dietary fiber and natural antioxidants to food products.

Winery by-products: extraction optimization, phenolic composition and cytotoxic evaluation to act as a new source of scavenging of reactive oxygen species.

Nearly 20 million tons of winery by-products, with many biological activities, are discarded each year in the world. The extraction of bioactive compounds from Chenin Blanc, Petit Verdot, and Syrah grape by-products, produced in the semi-arid region in Brazil, was optimized by a Central Composite Rotatable Design. The phenolic compounds profile, antioxidant capacity against synthetic free radicals (DPPH and ABTS), reactive oxygen species (ROS; peroxyl radical, superoxide radical, hypochlorous acid), cytotoxicity assay (MTT) and quantification of TNF-α production in RAW 264.7 cells were conducted. Gallic acid, syringic acid, procyanidins B1 and B2, catechin, epicatechin, epicatechin gallate, quercetin 3-ß-d-glucoside, delfinidin 3-glucoside, peonidin 3-O-glucoside, and malvidin 3-glucoside were the main phenolic compounds identified. In general, rachis showed higher antioxidant capacity than pomace extract, especially for Chenin Blanc. All extracts showed low cytotoxicity against RAW 264.7 cells and Petit Verdot pomace suppressed TNF-α liberation in vitro. Therefore, these winery by-products can be considered good sources of bioactive compounds, with great potential for application in the food and pharmaceutical industries.

Inhibition of DMBA-induced Oral Squamous Cells Carcinoma Growth by Brazilian Red Propolis in Rodent Model.

We investigated the effect of oral administration of hydroalcoholic extract of Brazilian red propolis (HERP) on DMBA-induced oral squamous cell carcinomas (OSCC) in rodents. The chemical components of the HERP were assessed by high-performance liquid chromatography (HPLC). Carcinogenesis was topically induced in the lower lip of 25 rats using 9,10-dimethyl-1,2-benzanthracene (DMBA); the tumour was treated with saline (TUM1) and Tween 80 (TUM2) as well as HERP at 10, 50 and 100 mg/kg (HERP10, HERP50 and HERP100, respectively) for 20 weeks. Topical application of saline and oral administration of 100 mg/kg HERP was used in five rats as a control group (CTR). After 26 weeks, the histological malignancy grading and immunohistochemical expression of Ki-67 and p16(INK4A) were assessed in the tumours/tissue samples. The compounds identified were propyl gallate, daidzein, catechin, epicatechin, formononetin and biochanin A. Formononetin, daidzein and biochanin A showed concentration of 23.29, 0.38 and 0.67 mg/g of HERP, respectively. HERP at doses of 50 and 100 mg/kg inhibited 40% of OSCC growth and promoted a 3-week delay in development of clinically detectable tumours. Epithelial dysplasia was observed in all samples with no clinical tumour, except in CTR. No significant difference in the immunoexpression of Ki-67 and p16(INK4A) was observed between HERP-treated and saline/Tween 80-treated groups (p > 0.05). Our results suggest that HERP exerts chemopreventive activity on the progression of DMBA-induced epithelial dysplasia to OSCC in an experimental model of labial carcinogenesis; however, this effect is not associated with Ki-67 and p16(INK4A) immunoexpression.

Composição fenólica e atividade antioxidante de resíduos agroindustriais / Phenolic composition and antioxidant activity of agroindustrial residues

Atualmente, são produzidas milhões de toneladas de resíduos provenientes do processamento agroindustrial. Muitos deles são ricos em compostos bioativos sendo potenciais fontes naturais dessas substâncias. Assim, este trabalho teve como objetivo avaliar o teor de compostos fenólicos totais, a atividade antioxidante e a composição fenólica de três resíduos gerados por agroindústrias brasileiras: bagaço de uva Isabel (BI) (Vitis labrusca), bagaço de uva Verdejo (BV) (Vitis vinifera) e bagaço de goiaba (BG) (Psidium guajava). Os resultados do teor de compostos fenólicos totais (mg GAE g-1) encontrados nos extratos etanólicos e aquosos dos resíduos foram, respectivamente: BV (20,94±0,46; 8,03±0,43)> BI (16,57±0,19; 4,41±0,01)> BG (3,41±0,09; 1,88±0,06). Alta atividade antioxidante, principalmente em BV e BI, foi verificada nos ensaios realizados (ABTS ●, DPPH ● e auto-oxidação do sistema beta-caroteno/ácido linoléico). Uma forte correlação positiva entre atividade antioxidante e o teor de compostos fenólicos totais foi encontrada. Os compostos fenólicos encontrados, por cromatografia gasosa com espectrometria de massas (CG-EM), foram: ácido gálico, epicatequina, quercetina (BV, BI e BG); ácido isovanílico (BI, BG); ácido p-cumárico (BI); ácido caféico e resveratrol (BV, BI). Esses resultados mostram que os resíduos agroindustriais analisados, particularmente os vinícolas, são ricos em substâncias bioativas e podem ser explorados pela indústria de alimentos e farmacêutica.

Nowadays, the agro-industrial processing produces millions of tons of wastes. Many of them are rich in bioactive compounds, being a potential natural source of these substances. This study aimed to evaluate the content of total phenolics, antioxidant activity and phenolic composition of residues generated by three Brazilian agribusiness: Isabel grape pomace (PI) (Vitis labrusca), Verdejo grape pomace (PV) (Vitis vinifera) and guava pomace (PG) (Psidium guajava). The results of total phenolics content (mg GAE g-1) found in the ethanol and aqueous extracts of residues were: PV (20.94±0.46; 8.03±0.43)>PI (16.57±0.19; 4.41±0.01)>PG (3.41±0.09; 1.88±0.06). High antioxidant activity of these extracts, particularly PV and PI, was found by the methods DPPH ●, ABTS ● and beta-carotene bleaching method. A strong positive correlation between antioxidant activity and content of total phenolic compounds was found. The following phenolic compounds were found by gas chromatography with mass spectrometry (GC-MS): gallic acid, epicatechin, quercetin (PV, PI and PG); isovanilic acid (PI, PG), p-coumaric acid (PI), caffeic acid and resveratrol (PV, PI). The results show that these residues, particularly the wineries, are rich in bioactive substances and should be exploited by the food industry and pharmaceuticals.