Antiproliferative Effect of Colonic Fermented Phenolic Compounds from Jaboticaba (Myrciaria trunciflora) Fruit Peel in a 3D Cell Model of Colorectal Cancer.

Jaboticaba is a Brazilian native berry described as a rich source of phenolic compounds (PC) with health promoting effects. PC from jaboticaba peel powder (JPP) have low intestinal bio-accessibility and are catabolized by gut microbiota. However, the biological implication of PC-derived metabolites produced during JPP digestion remains unclear. This study aimed to evaluate the antiproliferative effects of colonic fermented JPP (FJPP) in a 3D model of colorectal cancer (CRC) composed by HT29 spheroids. JPP samples fermented with human feces during 0, 2, 8, 24 or 48 h were incubated (10,000 µg mL-1) with spheroids, and cell viability was assessed after 72 h. Chemometric analyses (cluster and principal component analyses) were used to identify the main compounds responsible for the bioactive effect. The antiproliferative effect of FJPP in the CRC 3D model was increased between 8 h and 24 h of incubation, and this effect was associated with HHDP-digalloylglucose isomer and dihydroxyphenyl-γ-valerolactone. At 48 h of fermentation, the antiproliferative effect of FJPP was negligible, indicating that the presence of urolithins did not improve the bioactivity of JPP. These findings provide relevant knowledge on the role of colonic microbiota fermentation to generate active phenolic metabolites from JPP with positive impact on CRC.

Extraction, characterization and microencapsulation of isoflavones from soybean molasses / Extração, caracterização e microencapsulação de isoflavonas do melaço de soja

ABSTRACT: This study aimed to extract isoflavones from soybean molasses with different solvents, furthermore, the selected extract, which showed overall desirable characteristics was selected to evaluate the potentials of different encapsulating agents. The encapsulating agents employed for the study included 18% Maltodextrin DE20 (T1), 18% Hi-maize (T2), and a mixture of equal proportions of 9% Maltodextrin DE20 and 9% of Hi-maize (T3). Solvents such as 80% ethanol and methanol, and grain alcohol in varying different concentrations of 50 and 80% were used for the studies. The best solvent for the extraction of phenolics and total isoflavones was 50% cereal alcohol, this extract also presented higher antioxidant activity. Evaluation of the encapsulating agents revealed that 18% Hi-maize with inlet air of 130 °C was best suited for the encapsulation of isoflavones. The ORAC method showed that microcapsules with the 18% Hi-maize encapsulating agent also had higher antioxidant activity.

RESUMO: Este estudo teve como objetivo extrair isoflavonas do melaço de soja com diferentes solventes, além disso, o extrato selecionado com as características desejáveis ​​em geral foi selecionado para avaliar os potenciais de diferentes agentes encapsulantes. Os agentes encapsulantes foram: 18% de maltodextrina DE20 (T1), 18% Hi-maize (T2) e uma mistura em proporções iguais de 9 % de maltodextrina DE20 e 9% de Hi-maize (T3). Foram testados os solventes, etanol e metanol a 80% e álcool de cereais a 50 e 80%. O melhor solvente para a extração de fenólicos e isoflavonas totais foi o álcool de cereais a 50%, sendo que este extrato também apresentou maior atividade antioxidante. Entre os agentes encapsulantes testados, 18% Hi-maize com ar de entrada de 130 °C mostrou ser o melhor para encapsular as isoflavonas. O método ORAC mostrou que as microcápsulas com o agente encapsulante Hi-maize a 18% também apresentaram maior atividade antioxidante.

Grape peel powder promotes intestinal barrier homeostasis in acute TNBS-colitis: A major role for dietary fiber and fiber-bound polyphenols.

Inflammatory bowel diseases are characterized by impaired intestinal barrier function. This study aimed to evaluate the effects of grape peel powder (GPP) and its bioactive rich-fractions on the barrier function and colonic injury in a model of colitis induced by 2,4,6 trinitrobenzene sulfonic acid (TNBS). Wistar rats received diets supplemented with either GPP (8%), extractable polyphenols (EP), non-extractable polyphenols-rich fraction (NEP-F), or polyphenols-poor, fiber-rich fraction (F) from grapes at amounts equivalent to the GPP group during 15 days before and for 7 days after colitis induction. NEP-F has decreased the extension of colonic lesion but the other grape peel bioactive fractions did not protect against macroscopic or microscopic colonic damage, EP diet increased macroscopic colonic damage. GPP, EP, and NEP-F reduced claudin-2 mRNA expression, whereas GPP and F fraction increased occludin and ZO-1 mRNA expression. All experimental diets reduced the colitis-triggered increase of MMP-9 mRNA expression. Colitis reduced by 30% the production of cecal short-chain fatty acids (SCFA). GPP and NEP-F completely protected against this effect, whereas F fraction was ineffective. Only GPP and NEP-F were able to decrease the upregulation of GRP94 mRNA triggered by colitis. Dietary fiber seems to reestablish the intestinal barrier function, whereas fiber-bound phenolics were able to restore cecal metabolism to produce beneficial metabolites like SCFA and to reduce the activation of the unfolded protein response.

Analysis of polyphenols in brewer's spent grain and its comparison with corn silage and cereal brans commonly used for animal nutrition.

Brewer's spent grain (BSG) could be tested as an alternative source of polyphenols in animal nutrition. Proper extraction and analytical methods are critical for quantification. Thus, extraction for BSG, corn silage, and brans of rice, corn, and wheat were studied for the highest yield of polyphenols. A method for 18 phenolic monomers by HPLC-DAD was developed, validated, and applied to samples. An aqueous solution of NaOH (0.75% w/v) using integral samples for extraction resulted in the highest values for colorimetric measurements in all analyzed sources. Method by maceration showed the highest phenolic yield when applied in corn silage and BSG. However, for brans the best method was microwave assisted. Results from HPLC-DAD analysis clearly showed that native structures of phenolic compounds were simplified to its monomers allowing quantification and sample discrimination. BSG had the highest concentration of polyphenols and could be a promising and innovative source for animal feed studies.

Postharvest UV-C irradiation stimulates the non-enzymatic and enzymatic antioxidant system of 'Isabel' hybrid grapes (Vitis labrusca×Vitis vinifera L.).

Ultraviolet light type C (UV-C) was studied as a tool to increase enzymatic and non-enzymatic antioxidant defenses and phytochemical levels in 'Isabel' grapes (Vitis labrusca×Vitis vinifera L.). Grapes were exposed to 0, 0.5, 1.0, 2.0, or 4.0kJm-2 UV-C and stored for 1, 3, or 5days post-treatment. One day after UV-C irradiation, the activities of grape antioxidant enzymes and thiols were increased, especially at 1.0 and 2.0kJm-2. These doses increased total phenolic content by almost 20%, while 0.5 and 4.0kJm-2 had no effects. Total monomeric anthocyanin content was increased by >35% by UV-C at 1.0kJm-2; however, anthocyanin profile was unchanged. Grape skin antioxidant capacity was also improved by UV-C irradiation. The 1.0kJm-2 UV-C was considered the hormetic dose. Postharvest UV-C had an elicitor effect on 'Isabel' grapes, positively impacting the antioxidant capacity and phytochemical content.

Thermal Pest Control in 'Tannat' grapes: Effect on anthocyanins, sensory and color of one-year-old wines.

Thermal pest control (TPC) is a technology that applies heated air in the vineyard to reduce pests. However, it can also alter grape anthocyanin synthesis in answer to temperature stress. The aim of this work was to determine the impact of TPC in 'Tannat' grape vineyards on wine quality. The major makers of quality considered in this study were the wine anthocyanin characterization by HPLC-PDA-MSn, beverage's sensory evaluation by descriptive analysis, and color parameters by instrumental analysis. A total of three experiments were performed in different harvesting years. In 2011/12 (Experiment I), four applications were performed from the flowering to fruiting. For the second experiment (2012/13), five applications were used only during fruiting. TPC treatment performed in 2013/14 (Experiment III) had a total of 19 applications from the flowering to harvesting. In the first experiment, TPC had increased wine pigments, color, and sensorial descriptors for visual (translucence) and gustatory (woody and body) perceptions. However, in the second experiment none of the evaluated wine quality parameters was affected by TPC (p≤0.05). In the last and longest experiment (19 applications), TPC has significantly increased some wine pigments content. Nonetheless, wine sensorial parameters were not affected. Since the climate was different among the years, it seems that TPC effect does not depend on the applications number and vegetable cycle period only, but also it can be affected by the weather conditions in which the vine is being cultivated. However, when TPC treatment has resulted in the greatest anthocyanin increment (Experiment I) some wine sensory descriptors were also improved.

Effect of drying methods on the phenolic content and antioxidant capacity of Brazilian winemaking byproducts and their stability over storage.

This work aimed to study the antioxidant capacity by different methods, the total content of polyphenols and the stability over time of dried byproducts from Brazilian hybrids and Vitis vinifera varieties. Oven-dried at 50 °C and spray-dried samples were monitored for 90 days of storage. Under testing conditions, BRS Violeta grapes showed the greatest stability and initial high levels of total phenolics and anthocyanins remained almost unchanged until the end of storage period. The same behavior was observed in BRS Violeta freeze-dried skins, seeds and lees (8557, 9520 and 4261 mg GAE/100 g DM, respectively, and 829 and 257 mg mv-3-glc/100 g DM in skin and lees, respectively). In all methodologies tested, BRS Violeta also showed higher values for antioxidant capacity. These results suggest that dried winemaking byproducts can be used as rich sources of polyphenol compounds for industrial extractions with high stability and antioxidant capacity.

Biochemical Characterization of Dovyalis hebecarpa Fruits: A Source of Anthocyanins with High Antioxidant Capacity.

UNLABELLED: Dovyalis hebecarpa, which is also known as the Ceylon gooseberry, is an attractive exotic purple berry that is produced in the southwest of Brazil with high yields. Current literature lacks information about the nutritional composition and antioxidant capacity of this fruit. As such, this work aimed to evaluate and compare the proximate composition, phytochemical content, and antioxidant capacity of D. hebecarpa over two seasons through the application of a range of different methods, including FRAP, ABTS, and ORAC assays. The proximate composition and biometric parameters were monitored for quality purposes. The results significantly changed between seasons in accordance with variations in rain incidence and average temperatures. The Ceylon gooseberry contained a similar or higher content of anthocyanins (ACY) than other berries commonly consumed in human diet. These ACY were identified at higher levels in the fruit's skin (284 to 351 mg of cyanidin-3-glucoside equivalents/100 g of fresh weight [FW]) than in its pulp (49 to 69 mg/100 g of FW). The use of an ORAC assay identified higher antioxidant activity (145 µmol of Trolox equivalents [TE]/g in the skins and 179 µmol of TE/g in the pulp on a FW basis) than all other methods used to determine antioxidant activity. Thus, D. hebecarpa is a rich source of ACY that demonstrates high antioxidant activity against the peroxyl radicals formed in ORAC assay conditions. PRACTICAL APPLICATION: Ceylon gooseberry fruit is a source of phytochemicals that could be used in the human diet providing defense against free radicals damage. Moreover, the skins of the fruit, which are not typically consumed, contained higher levels of polyphenols than the pulp. This indicates that the skins of the Ceylon gooseberry may represent a promising source of natural pigments and antioxidants for industrial applications.

Anthocyanin and other phenolic compounds in Ceylon gooseberry (Dovyalis hebecarpa) fruits.

Ceylon gooseberry is a deep-purple exotic berry that is being produced in Brazil with great market potential. This work aimed to determine major phenolic compounds in this specie by HPLC-PDA-ESI/MS. Samples were collected in two different seasons. Pulp and skin were analyzed separately. Non-acylated rutinoside derivatives of delphinidin (∼60-63%) and cyanidin (∼17-21%) were major anthocyanins tentatively identified. All anthocyanins had higher concentration in skin than in pulp (64-82 and 646-534mg of cyaniding-3-glucoside equivalents/100g skin and pulp, respectively). Moreover, anthocyanin profile changed between sampling dates (p<0.05). Mainly for delphinidin-3-rutinoside which could be a result of season variation. In this specie, non-anthocyanin polyphenols represent less than 35% of total extracted polyphenols. The tentative identification proposed a flavonol and three ellagitannins as major compounds of the non-anthocyanin phenolics fraction. Finally, anthocyanin is the major phenolic class in this fruit and its composition and content are significantly affected by season.

Polyphenol extraction optimisation from Ceylon gooseberry (Dovyalis hebecarpa) pulp.

Originally from Asia, Dovyalis hebecarpa is a dark purple/red exotic berry now also produced in Brazil. However, no reports were found in the literature about phenolic extraction or characterisation of this berry. In this study we evaluate the extraction optimisation of anthocyanins and total phenolics in D. hebecarpa berries aiming at the development of a simple and mild analytical technique. Multivariate analysis was used to optimise the extraction variables (ethanol:water:acetone solvent proportions, times, and acid concentrations) at different levels. Acetone/water (20/80 v/v) gave the highest anthocyanin extraction yield, but pure water and different proportions of acetone/water or acetone/ethanol/water (with >50% of water) were also effective. Neither acid concentration nor time had a significant effect on extraction efficiency allowing to fix the recommended parameters at the lowest values tested (0.35% formic acid v/v, and 17.6 min). Under optimised conditions, extraction efficiencies were increased by 31.5% and 11% for anthocyanin and total phenolics, respectively as compared to traditional methods that use more solvent and time. Thus, the optimised methodology increased yields being less hazardous and time consuming than traditional methods. Finally, freeze-dried D. hebecarpa showed high content of target phytochemicals (319 mg/100g and 1,421 mg/100g of total anthocyanin and total phenolic content, respectively).

Diferentes fontes protéicas para a alimentacão do jundiá (Rhamdia quelen) / Different protein sources for jundiá (Rhamdia quelen) feeding

O objetivo deste trabalho foi verificar o crescimento e a composicão de filés de juvenis de jundiá (Rhamdia quelen) alimentados com diferentes fontes protéicas, durante 60 dias. Utilizaram-se 540 peixes (peso médio inicial=15,00n0,62g, comprimento total inicial=11,98n0,35cm) distribuídos ao acaso em 18 caixas de 280L (30 peixes/caixa) em sistema de recirculacão de água. Testaram-se seis tratamentos (com três repeticões): CL (farinha de carne e ossos + levedura), SL (farelo de soja + levedura), S (somente farelo de soja), CS (farinha de carne e ossos + farelo de soja), PL (farinha de peixe + levedura) e PS (farinha de peixe + farelo de soja). Foram estimados parâmetros de desempenho (peso, comprimento total, fator de condicão, taxa de crescimento específico, taxa de eficiência protéica) aos 30 e 60 dias, composicão centesimal e taxas de deposicão de proteína e gordura nos filés ao final do experimento. Os parâmetros peso, comprimento total e taxa de crescimento específico foram superiores nos tratamentos compostos pela combinacão das farinhas de origem animal (carne e ossos e peixe) com farelo de soja. A quantidade de gordura e proteína depositada no filé dos peixes também foi superior nestes tratamentos (CS e PS). Os rendimentos de carcaca e filé não diferiram entre os tratamentos. A combinacão das farinhas de carne e ossos e de peixes com farelo de soja possibilita bom crescimento e maior deposicão de proteína e gordura nos filés de juvenis de jundiá.