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In order to contribute to improve knowledge about the actions of Camellia sinensis extracts on starch digestion, several varieties were compared. The latter were green, oolong, white, black, and purple teas. The results are hoped to contribute to our understanding of the mode of action and potency of the various tea preparations as possible adjuvants in the control of post-prandial glycemia. The extracts were prepared in way similar to their form of consumption. All extracts decreased starch digestion, but the purple tea extract was the strongest inhibitor, their inhibitory tendency started at the dose of 50 mg/kg and was already maximal with 250 mg/kg. Maltose tolerance was not significantly affected by the extracts. Glucose tolerance was not affected by purple tea, but black tea clearly diminished it; green tea presented the same tendency. Purple tea was also the strongest inhibitor of pancreatic α-amylase, followed by black tea. The green tea, oolong tea, and white tea extracts tended to stimulate the pancreatic α-amylase at low concentrations, a phenomenon that could be counterbalancing its inhibitory effect on starch digestion. Based on chemical analyses and molecular docking simulations it was concluded that for both purple and black tea extracts the most abundant active component, epigallocatechin gallate, seems also to be the main responsible for the inhibition of the pancreatic α-amylase and starch digestion. In the case of purple tea, the inhibitory activity is likely to be complemented by its content in deoxyhexosidehexoside- containing polyphenolics, especially the kaempferol and myricetin derivatives. Polysaccharides are also contributing to some extent. Cyanidins, the compounds giving to purple tea its characteristic color, seem not to be the main responsible for its effects on starch digestion. It can be concluded that in terms of postprandial anti-hyperglycemic action purple tea presents the best perspectives among all the tea varieties tested in the present study. This work was financially supported by grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-304090/ 2016-6), Coordenação do Aperfeiçoamento de Pessoal do Ensino Superior (CAPES) and Fundação Araucária (53/2019). The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020) and through the institutional scientific employment program-contract for L. Barros and M.I. Dias contracts. This work was also funded by the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project Mobilizador ValorNatural®: Norte-01-0247-FEDER-024479 and GreenHealth: Norte-01-0145-FEDER-000042. info:eu-repo/semantics/publishedVersion

Triclosan (5-chloro-2'-[2,4-dichlorophenoxi]-phenol) is a polychlorinated biphenolic antimicrobial, utilized as antiseptic and preservative in hygiene products and medical equipment. Triclosan causes mitochondrial dysfunction (uncoupling, inhibition of electron flow), as demonstrated in isolated rat liver mitochondria. These actions in the mitochondria could compromise energy-dependent metabolic fluxes in the liver. For this reason, the present work aimed at investigating how these effects on isolated mitochondria translate to the whole and intact hepatocyte. For accomplishing this, the isolated perfused rat liver was utilized, a system that preserves both microcirculation and the cell-to-cell interactions. In addition, the single-pass triclosan hepatic transformation was also evaluated by HPLC as well as the direct action of triclosan on gluconeogenic enzymes. The results revealed that triclosan decreased anabolic processes (e.g., gluconeogenesis) and increased catabolic processes (e.g., glycolysis, ammonia output) in the liver, generally with a complex pattern of concentration dependences. Unlike the effects on isolated mitochondria, which occur in the micromolar range, the effects on intact liver required the 10

The actions of resveratrol in brain and plasma of rats with adjuvant-induced arthritis were investigated. Resveratrol was administered orally during a period of 23 days. A major concern of the present work was to explore an ample range of daily doses (10-200 mg/kg). Several oxidative and inflammatory markers were measured. Important effects of resveratrol treatment were the normalization of the plasma myeloperoxidase activity (inflammatory marker), the normalization of the brain xanthine oxidase activity (reactive oxygen species source) and the near-normalization of the catalase activity in the brain (antioxidant defence). These effects presented obvious dose dependencies in the range up to 200 mg/kg. Resveratrol also reduced protein and lipid damage within the lowest dose ranges investigated, and its action as a free radical scavenger activity was enhanced in brain mitochondria of arthritic rats. Resveratrol failed in restoring the diminished albumin levels and plasma protein thiols in arthritic rats. The latter, however, were substantially increased in healthy rats at low doses (up to 50 mg/kg), a sign of antioxidant action. This increase was reversed at higher doses, a sign of pro-oxidant action. The observations agree with the notion that low doses of resveratrol might be useful as an adjuvant to the conventional antirheumatic drugs.

Cerebral ischemia-induced hyperglycemia has been reported to accentuate neurological damage following focal or global cerebral ischemia. Hyperglycemia found in rats following focal brain ischemia occurs in the first 24 h and has been claimed to be caused by increased liver gluconeogenesis and insulin resistance. However, liver gluconeogenesis and the mechanisms leading to hyperglycemia after global cerebral ischemia remain uncertain. This study investigated the glycemic homeostasis and hepatic metabolism in rats after transient four-vessel occlusion (4-VO)-induced global cerebral ischemia, an event that mimics to a certain degree the situation during cardiac arrest. Several metabolic fluxes were measured in perfused livers. Activities and mRNA expressions of hepatic glycolysis and glyconeogenesis rate-limiting enzymes were assessed as well as respiratory activity of hepatic isolated mitochondria. Global cerebral ischemia was associated with hyperglycemia and hyperinsulinemia 24 h after ischemia. Insulin resistance developed later and was prominent after the 5th day. Hepatic anabolism and catabolism were both modified in a complex and time-dependent way. Gluconeogenesis, β-oxidation, ketogenesis and glycolysis were diminished at 24 h after ischemia. At 5 days after ischemia glycolysis had normalized, but gluconeogenesis, ketogenesis and β-oxidation were accelerated. The overall metabolic modifications suggest that a condition of depressed metabolism was established in response to the new conditions generated by the cerebral global ischemia. Whether the modifications in the liver metabolism found in rats after the ischemic insult can be translated to individuals following global brain ischemia remains uncertain, but the results of this study are hoped to encourage further investigations.

This work aimed to provide information that contributes to establishing environmental-friendly methods for synthetic dyes' degradation. The potential decolorization capacity of the crude enzymatic extract produced by Phanerochaete chrysosporium CDBB 686 using corncob as a substrate was evaluated on seven different dyes. Critical variables affecting the in-vitro decolorization process were further evaluated and results were compared with an in-vivo decolorization system. Decolorization with enzymatic extracts presented advantages over the in-vivo system (higher or similar decolorization within a shorter period). Under improved in-vitro process conditions, the dyes with higher decolorization were: Congo red (41.84 %), Poly R-478 (56.86 %), Methyl green (69.79 %). Attempts were made to confirm the transformation of the dyes after the in-vitro process as well as to establish a molecular basis for interpreting changes in toxicity along with the degradation process. In-vitro degradation products of Methyl green presented a toxicity reduction compared with the original dye; however, increased toxicity was found for Congo red degradation products when compared with the original dyes. Thus, for future applications, it is crucial to evaluate the mechanisms of biodegradation of each target synthetic dye as well as the toxicity of the products obtained after enzymatic oxidation.

A practical approach to control glycemia in diabetes is to use plant natural products that delay hydrolysis of complex sugars and promote the diminution of the release of glucosyl units into the blood plasma. Polyphenolics have been described as being effective in inhibiting amylases and α-glucosidases. Grape pomace is an important sub product of the wine industry, still rich in many compounds such as polyphenolics. In this context, the purpose of this study was to search for possible effects of a grape pomace extract on salivary and pancreatic α-amylases and α-glucosidase, as well as on intestinal glucose absorption. The Merlot grape pomace extract (MGPE) was prepared using a hydroalcoholic mixture (40% ethanol + 60% water). In vitro inhibition was quantified using potato starch (for amylases) and maltose (for α-glucosidase) as substrates. In vivo inhibition was evaluated by running starch and maltose tolerance tests in rats with or without administration of MGPE. Ranking of the extract compounds for its affinity to the α-amylases was accomplished by computer simulations using three different programs. Both α-amylases, pancreatic and salivary, were inhibited by the MGPE. No inhibition on α-glucosidase, however, was detected. The IC50 values were 90 ± 10 μg/mL and 143 ± 15 μg/mL for salivary and pancreatic amylases, respectively. Kinetically this inhibition showed a complex pattern, with multiple binding of the extract constituents to the enzymes. Furthermore, the in silico docking simulations indicated that several phenolic substances, e.g., peonidin-3-O-acetylglucoside, quercetin-3-O-glucuronide and isorhamnetin-3-O-glucoside, besides catechin, were the most likely polyphenols responsible for the α-amylase inhibition caused by MGPE. The hyperglycemic burst, an usual phenomenon that follows starch administration, was substantially inhibited by the MGPE. Our results suggest that the MGPE can be adequate for maintaining normal blood levels after food ingestion. The authors wish to thank to the Fundação Araucária (Brazil), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brazil), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil), Cesumar Institute of Science Technology and Innovation (ICETI, Brazil), and FEDER-Interreg España-Portugal for their financial help. info:eu-repo/semantics/publishedVersion

The aqueous extracts of mycelium (MAE) and basidioma (BAE) of the mushroom Pleurotus pulmonarius (Fr.) Quel. were compared for their chemical composition, phenolic compounds identified using HPLC and antioxidant activities. The MAE contained 66.1 μg of gallic acid, 27.0 μg of ferulic acid and 217 μg of caffeic acid/g dry weight (dw). In addition, 626 μg of gallic acid, 347 μg of caffeic acid, 53.0 μg of vanillic acid and 25.4 μg of ρ-coumaric acid were found in BAE/g dw. The levels of soluble proteins, soluble carbohydrates and free amino acids were higher in the BAE. The MAE of P. pulmonarius were efficient in chelating ferrous ions (EC50 of 1.81 mg dw/mL). The EC50 for DPPH and ABTS radicals were 0.51 and 0.03 mg dw/mL, respectively, showing high antioxidant capacity. The results of this study showed that mycelium and basidioma of P. pulmonarius have the potential to be used as sources of antioxidant compounds and nutrients.

Additional file 1: Table S1. Software scores found on docking ligands to porcine pancreatic alpha amylase.

A study of the effects of butylated hydroxyanisole (BHA) on the hepatic metabolism was conducted with emphasis on parameters linked to energy metabolism and mitochondrial reactive oxygen species production. The experimental systems were the isolated perfused rat liver and isolated mitochondria. It was found that BHA inhibits biosynthetic pathways (gluconeogenesis) and ammonia detoxification, which are dependent on ATP generated within the mitochondria. Conversely, the compound stimulated glycolysis and fructolysis, which are compensatory phenomena for an inhibited mitochondrial ATP generation. Furthermore, BHA diminished the cellular ATP content under conditions where the mitochondrial respiratory chain was the only source of this compound. Inhibition of gluconeogenesis started at the concentration of 50 μM and was generally pronounced at concentrations under 200 μM. Several effects, however, were prominent only at the concentrations of 500 and 750 μM. BHA can be considered, thus, a mild metabolic agent that becomes toxic only at high doses. An aggravating factor could be the observation that BHA exerts a net stimulating action on reactive oxygen species (ROS) production in isolated mitochondria, an observation that contradicts the general notion that the compound acts primarily as an antioxidant. Considerable time was required for the reversion of most effects after removal of the compound from the circulation. In toxicological terms, besides the lack of circulating glucose, one can expect metabolic acidosis due to excess lactate production, impairment of ammonia detoxification and cell damage due to a deficient maintenance of its homeostasis and possible excessive ROS production.

Ozonolysis is potentially an effective method for pretreating lignocellulosic biomass to improve the production of fermentable sugars via enzymatic hydrolysis. The eliminated branches from eucalyptus trees can represent a production of around 30 million m3 of lignocellulosic material annually only in Brazil. Attempts of developing strategies for a rational use of this biomass are, thus, welcome. In this study, Eucalyptus grandis branches were pretreated with ozone in an attempt to increase enzymatic saccharification. Ozonolysis resulted in the degradation of lignin with negligible losses of cellulose and small losses of hemicellulose. Reduction in the lignin content from 26.63 to 9.53% already resulted in the maximal improvement of the saccharification yield (from 20 to 68%). The results indicate that ozone pretreatment can be a promising way of increasing the enzymatic digestibility of eucalyptus sawdust from eliminated branches of trees for its conversion into fermentable sugars.

Endophytic and phytopathogenic isolates of the genus Fusarium were evaluated for their potential to produce cellulolytic and xylanolytic enzymes. The fungi were grown under solid-state conditions using a mixture of sugarcane bagasse and wheat bran as substrate with an initial moisture of 82%. Endoglucanases (ranging from 0.20 ± 0.03 to 5.31 ± 0.30 U/g of substrate), xylanases (ranging from 4.65 ± 0.76 to 125.57 ± 8.25 U/g of substrate), β-glucosidases (ranging from 21.48 ± 3.70 to 527.17 ± 22.14 U/g of substrate) and β-xylosidases (ranging from 5.61 ± 1.25 to 40.69 ± 1.26 U/g of substrate) were produced by all isolates. Enzymatic extracts from the best xylanase producers, F. lateritum var. majus and F. sacchari var. subglutinans, were tested for their capacities in promoting saccharification of delignified sugarcane bagasse. Amounts of 200.60 ± 10.60 mg/g and 280.00 ± 19.00 mg/g of reducing sugars, mainly xylose, were obtained after enzymatic hydrolysis by F. lateritum var. majus and F. sacchari var. subglutinans extracts, respectively. Up to now these species of Fusarium have been barely explored as sources of xylanolytic enzymes capable to degrade plant biomass, but our results open perspectives for their biotechnological use in the obtainment of xylose from delignified sugarcane bagasse.

The white rot basidiomycete Ganoderma lucidum was evaluated for its capability to tolerate and to degrade the herbicide diuron. Diuron at a subtoxic concentration was added at the start of the cultivation in glucose liquid stationary cultures. Under this condition diuron was a laccase inducer. Almost 50% of the initially present diuron was removed after 15 d of cultivation. Two diuron metabolites were found N'-(3,4-dichlorophenyl)-N-methylurea (DCPMU) and 3,4-dichlorophenylurea (DCPU). The addition of the cytochrome P450 inhibitors 1-aminobenzotriazole and piperonyl butoxide reduced significantly the capability of the fungus in degrading diuron. The activities of superoxide dismutase and catalase were significantly increased in the mycelial extracts by the presence of diuron. On the other hand, diuron did not cause any significant alteration in the levels of reactive oxygen species. Additionally, laccase could also degrade diuron in vitro and this degradation was increased by the addition of synthetic mediators, 3-ethylbenzthiazoline-6-sulphonic acid and acetylacetone. Significant reduction in the toxicity, as evaluated by the Lactuca sativa bioassay, was observed after G. lucidum treatment. In conclusion, G. lucidum is able to metabolize diuron by intra- and extracellular mechanisms, without the accumulation of toxic products.

Pentagalloyl-glucose (PGG) occurs in many plants and possesses several therapeutic properties, including anti-diabetic ones. Based on the latter, the inhibitory action of PGG on α-amylases was investigated. PGG inhibited both the pancreatic and the salivary α-amylases. With starch as the substrate, complexation with the free enzyme form either predominates (pancreatic enzyme) or is the only complexation that is effectively allowed (salivary enzyme; competitive inhibition). This contrasts with a previous report on the salivary enzyme showing non-competitive inhibition when a synthetic substrate was used. PGG was less effective than acarbose as an inhibitor of α-amylases and presented a different binding pattern in terms of the aminoacyl residues involved. Finally, PGG also diminished the hyperglycemic excursion normally caused by starch administration when it was given to rats simultaneously to the polysaccharide. This suggests that PGG or foods and infusions rich in PGG could be helpful as adjuvants for maintaining normal glycemia.

Background TNBS-induced colitis is an experimental immunopathology in rats that shares many features with human inflammatory bowel diseases. Copaiba oleoresin is extracted from plants of the genus Copaifera and is shown to reduce inflammation. Objective The aim of this study was to investigate the action of copaiba oil (C. reticulata Ducke) on inflammation and oxidative status in the distal colon of colitic rats. Methods Acute and subchronic colitis were induced in Wistar rats by an intracolonic enema with 2,4,6-trinitrobenzenesulfonic acid (TNBS). The colonic morphology was assessed by histological analysis and the oxidative stress parameters were measured in the intestinal homogenate. The liver damage markers were measured in the plasma. Control and colitic rats were orally treated either with one single dose (acute colitis) of copaiba oil (1.15 g Kg-1) or once a day during seven days (subchronic colitis). Results The intestinal morphology was severely modified by acute and subchronic colitis, as indicated by the intramural infiltration of polymorphonuclear cells and the increased thickness of all colon layers. The levels of TBARS, protein carbonyl groups and reactive oxygen species (ROS) were increased in the intestine of colitic rats. Copaiba oil did not attenuate the inflammatory damage in acute and subchronic colitis, but it decreased the activity of myeloperoxidase, leukocyte infiltration and oxidative stress in the colon. The level of plasma bilirubin and the activity of alkaline phosphatase were both increased in treated healthy and colitic rats. Conclusion Copaiba oil decreased oxidative stress and inflammation but did not prevent intestinal damage in the colon of colitic rats. The alterations of plasma markers of hepatic damage caused by the oil seem to be associated to its harmful action on the liver.

The purpose of this study was to evaluate possible effects of the administration of an aqueous Agaricus brasiliensis extract on the oxidative state of the liver, brain, and plasma in adjuvant-induced arthritic rats, a model for human rheumatoid arthritis. Daily doses of 400 mg · kg-1 were administered by gavage for 23 days. This treatment produced significant diminutions in protein carbonylation and lipid damage in the liver, brain, and plasma. It also diminished the tissue reactive oxygen species and increased the antioxidant capacity of the plasma. Antioxidant defenses, which are diminished by arthritis, were improved by treatment with the A. brasiliensis extract, as revealed by preservation of the reduced glutathione and protein thiol levels and by the tendency of the activities of some antioxidant enzymes to normalize. The increased glucose-6-phosphate dehydrogenase activity was also almost normalized by the treatment. In addition, there were indications that A. brasiliensis can inhibit the initiation of structural events that can lead to hepatocyte necrosis. In conclusion, A. brasiliensis aqueous preparations can, in principle, be visualized as potential auxiliaries in the treatment of patients with rheumatoid arthritis as a result of their capacity to reduce oxidative stress. This effect was exerted in multiple organs, as can be judged from the results obtained in the liver, brain, and plasma. The continuous ingestion of A. brasiliensis as specific preparations or as a food supplement can possibly help to attenuate the systemic effects of rheumatoid arthritis and improve the quality of life of patients affected by this disease.

Aims Quercetin has been investigated as an agent to treat rheumatoid arthritis. At high doses it improves inflammation and the antioxidant status of arthritic rats, but it also exerts mitochondriotoxic and pro-oxidant activities. Beneficial effects of quercetin have not been found at low doses because of its chemical instability and low bioavailability. In the hope of overcoming these problems this study investigated the effects of long-term administration of quercetin-loaded pectin/casein microparticles on the oxidative status of liver and brain of rats with adjuvant-induced arthritis. Main methods Particle morphology was viewed with transmission electron microscopy and the encapsulation efficiency was measured indirectly by X-ray diffraction. Quercetin microcapsules (10 mg/Kg) were orally administered to rats during 60 days. Inflammation indicators and oxidative stress markers were measured in addition to the respiratory activity and ROS production in isolated mitochondria. Key findings Quercetin was efficiently encapsulated inside the polymeric matrix, forming a solid amorphous solution. The administration of quercetin microparticles to arthritic rats almost normalized protein carbonylation, lipid peroxidation, the levels of reactive oxygen species as well as the reduced glutathione content in both liver and brain. The paw edema in arthritic rats was not responsive, but the plasmatic activity of ALT and the mitochondrial respiration were not affected by quercetin, indicating absence of mitochondriotoxic or hepatotoxic actions. Significance Quercetin-loaded pectin/casein microcapsules orally administered at a low dose improve oxidative stress of arthritic rats without a strong anti-inflammatory activity. This supports the long-term use of quercetin as an antioxidant agent to treat rheumatoid arthritis.

Bisphenol A (BPA) is an endocrine disruptor compound, that is continuously released into the environment and is barely degraded in wastewater treatment plants. In this work, crude laccases from Pleurotus ostreatus and Pleurotus pulmonarius were used for the first time to degrade BPA in aqueous solution. The residual BPA was quantified by HPLC and the BPA metabolites produced by action of the laccases were identified by GC–MS. BPA at 100 mg/L and 200 mg/L (0.88 mmol/L) were 100% and 85% removed, respectively, in a 1 h reaction by both fungal laccases at a concentration of 8 U/L. Thirteen aromatic and aliphatic BPA metabolites were identified, including p-isopropenylphenol, methylpent-3-oic acid, ethyl-3-ethoxy propanoate, and 4-ethyl-2-methoxyphenol. The acute BPA toxicity decreased from 85% to less than 5% using the P. ostreatus laccase. On the other hand, P. pulmonarius laccase, did not caused reduction in toxicity, possibly because at least one BPA metabolite was as toxic as the parent compound itself. The results of this study suggest a feasible method for the complete removal of BPA from polluted environments using crude laccase from P. ostreatus.

Background Consumption of foods enriched with phytosterols (PS) can promote substantial reductions in the low-density lipoprotein cholesterol concentrations, a major risk factor for cardiovascular diseases (CVD). For reaching the 2 g/d threshold dose for cholesterol-lowering purposes, the ingestion of PS-enriched foods as well as PS-based nutraceuticals is required. With a growing global functional food market moving billions every year, there are promising opportunities for the development of health claim-bearing sterol-enriched food products. Scope and approach The present review aims not only to summarize and critically discuss the research produced during the last decade on PS, but also to propose a natural, interesting and sustainable alternative to these compounds. For this purpose, this work presents the mycosterols as promising and still underexplored sources of functional ingredients for the development of sterol-enriched food products and dietary supplements. Key findings and conclusions Ergosterol-enriched fungi extracts are potential sources of biomolecules that could not only impair cholesterol absorption but also inhibit its biosynthesis in human body, besides promoting other significant health benefits such as anti-cancer, antiproliferative, anti-inflammatory and antimicrobial effects. In addition, as a precursor of Vitamin D2, ergosterol might improve bone metabolism, immunity and mood. The isolation of novel bioactive mycosterols from still underexploited fungi species, the optimization of the existing methodologies for production and isolation of these target mycochemicals (mainly automatized liquid fermentation technologies), as well as the extensive study of their application in food product formulations (including products' efficacy, stability, nutritional and sensorial evaluation), are current challenges for research.

Grape pomace is an agro-industrial residue produced worldwide. The purpose of this study was to identify and quantify the main phenolics present in an hydroalcohlic extract of a Merlot grape pomace and to investigate its effect on the oxidative and inflammatory states of adjuvant-induced arthritic rats. Daily doses of 250 mg of the extract per kg body weight were administered during 23 days. Several oxidative stress indicators in arthritic rats were maintained at their normal or closely normal levels in the plasma, liver and brain by the treatment. Additionally, the grape pomace also showed significant anti-inflammatory effects. From the 25 phenolics identified in the grape pomace extract the most abundant ones were catechin and catechin derivatives, which are possibly the most important antioxidant agents. The results suggest a potential applicability of the Merlot grape pomace hydroalcoholic extract in the improvement of the oxidative and inflammatory states in arthritic patients. The authors which to thank Anacharis Babeto de Sá-Nakanishi, Jurandir Fernando Comar and Leomara Floriano Ribeiro for their expert technical assistance. This work was financially supported by grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Proc. 302615/2011-3 and Proc. 307944/2015-8) and Fundação Araucária (Proc. 215/2014). Andréia A. Soares and Rúbia C. G. Corrêa were post-doc fellowship recipients of the Coordenação do Aperfeiçoamento de Pessoal do Ensino Superior (CAPES) and Fundação Araucária. The authors are also thankful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Program PT2020 for financial support to CIMO (UID/AGR/00690/2013) and L. Barros (SFRH/BPD/107855/2015) grant. To POCI-01-0145-FEDER-006984 (LA LSRE-LCM), funded by ERDF, through POCI-COMPETE2020 and FCT. info:eu-repo/semantics/publishedVersion

In this work, liquid nitrogen was used for the first time in the pretreatment of plant biomasses for purposes of enzymatic saccharification. After treatment (cryocrushing), the initial rates of the enzymatic hydrolysis of eucalyptus sawdust and rice hull were increased more than ten-fold. Cryocrushing did not modify significantly the contents of cellulose, hemicellulose and lignin in both eucalyptus sawdust and rice hulls. However, substantial disorganization of the lignocellulosic materials in consequence of the pretreatment could be observed by electron microscopy. Cryocrushing was highly efficient in improving the saccharification of the holocellulose component of the plant biomasses (from 4.3% to 54.1% for eucalyptus sawdust and from 3.9% to 40.6% for rice hull). It is important to emphasize that it consists in a simple operation with low requirements of water and chemicals, no corrosion, no release of products such as soluble phenolics, furfural and hydroxymethylfurfural and no waste generation.

p-synephrine and p-octopamine were found to increase lipolysis in adipocytes. The present study approaches the question if these compounds, natural products of the bitter orange (Citrus aurantium fruit), increase lipolysis and fatty acid oxidation in the liver. Experiments were done in the perfused rat liver. Non-recirculating hemoglobin-free perfusion was done using the Krebs/ Henseleit-bicarbonate buffer (pH 7.4) as perfusion fluid. Both p-synephrine and p-octopamine, at the concentrations of 100 μM, were found to stimulate the hepatic triacylglycerol lipase by 40% and 51%, respectively. These seem to be the maximal stimulations possible in the liver. In the perfused liver, p-synephrine, when present at an initial concentration of 500 μM, was able to increase the non-esterified fatty acid release after one hour of recirculating perfusion. The effects of p-synephrine on the oxidation of exogenously supplied [1-14C]octanoate and [1-14C]oleate were minimal. Only oxygen uptake, already stimulated by octanoate or oleate, was additionally increased by the infusion of p-synephrine. These results contrast with those obtained in a previous study with p-octopamine, which increased the production of 14CO2 from both [1-14C]octanoate and [1-14C]oleate. Apparently only the oxidation of endogenous fatty acids is stimulated by p-synephrine. On the other hand, both p-synephrine and p-octopamine stimulate the hepatic triacylglycerol lipase to a much lesser extent than the adipocyte lipase. It can be concluded that p-synephrine affects much more carbohydrate metabolism in the liver than lipid metabolism.

Methyl jasmonate (MeJA), common in the plant kingdom, is capable of reducing articular and hepatic inflammation and oxidative stress in adjuvant-induced arthritic rats. This study investigated the actions of orally administered MeJA (75&ndash, 300 mg/kg) on inflammation, oxidative stress and selected enzyme activities in the brain of Holtzman rats with adjuvant-induced arthritis. MeJA prevented the arthritis-induced increased levels of nitrites, nitrates, lipid peroxides, protein carbonyls and reactive oxygen species (ROS). It also prevented the enhanced activities of myeloperoxidase and xanthine oxidase. Conversely, the diminished catalase and superoxide dismutase activities and glutathione (GSH) levels caused by arthritis were totally or partially prevented. Furthermore, MeJA increased the activity of the mitochondrial isocitrate dehydrogenase, which helps to supply NADPH for the mitochondrial glutathione cycle, possibly contributing to the partial recovery of the GSH/oxidized glutathione (GSSG) ratio. These positive actions on the antioxidant defenses may counterbalance the effects of MeJA as enhancer of ROS production in the mitochondrial respiratory chain. A negative effect of MeJA is the detachment of hexokinase from the mitochondria, which can potentially impair glucose phosphorylation and metabolism. In overall terms, however, it can be concluded that MeJA attenuates to a considerable extent the negative effects caused by arthritis in terms of inflammation and oxidative stress.

Tordon® is the commercial name of a mixture of two organo-chlorinated herbicides, 2,4-D and picloram. Both compounds affect energy transduction in isolated mitochondria and the present study aimed at characterizing the actions of these two compounds on liver metabolism and their cellular distribution in the isolated perfused rat liver. 2,4-D, but not picloram, increased glycolysis in the range from 10 to 400 μM. The redox potential of the cytosolic NAD+-NADH couple was also increased by 2,4-D. Both compounds inhibited lactate gluconeogenesis. Inhibitions by 2,4-D and picloram were incomplete, reaching maximally 46% and 23%, respectively. Both compounds diminished the cellular ATP levels. No synergism between the actions of 2,4-D and picloram was detected. Biotransformations of 2,4-D and picloram were slow, but their distributions occurred at high rates and were concentrative. Molecular dynamics simulations revealed that 2,4-D presented low affinity for the hydrophobic lipid bilayers, the opposite occurring with picloram. Inhibition of energy metabolism is possibly a relevant component of the toxicity of 2,4-D and of the commercial product Tordon®. Furthermore, the interactions of 2,4-D with the membrane lipid bilayer can be highly destructive and might equally be related to its cellular toxicity at high concentrations.

The food manufacturing industry has increasingly focused in the development of wholesome and safer products, including certified labeled “super foods,” “healthy foods” and “functional foods,” which are currently under great demand worldwide. Plant pigments and vitamins are amidst the most common additives incorporated to foodstuff, not only for improving their nutritional status but also for coloration, preservation, and even therapeutic purposes. The recovery of pigments from agro industrial wastes using green emerging approaches is a current trend and clearly the best alternative to ensure their sustainable obtainment and make these ingredients more popular, although still full of challenging aspects. Stability and bioavailability limitations of these active molecules in food matrices have been increasingly studied, and a number of methods have been proposed to minimize these issues, among which the incorporation of a co-pigment, exclusion of O2 during processing and storage, and above all, microencapsulation and nanoencapsulation techniques. The most recent advances and challenges in the application of natural pigments and vitamins in functional foods, considering only reports of the last 5 years, were the focus of this chapter.

Response surface methodology was applied to optimize the recovery of ergosterol from an agroindustrial residue of Agaricus blazei Murrill, by comparing conventional (heat-assisted extraction, HAE) and more sustainable nonconventional approaches (ultrasound and microwave-assisted extractions, UAE and MAE, respectively). A circumscribed central composite design was employed to optimize and describe the interactive effects among variables of the HAE, MAE (time and temperature) and UAE systems (time and ultrasound power), and their performances compared with the standard soxhlet extraction system. The optimum conditions were predicted as 150 min, 81.6 °C for HAE, 30 min, 400W for UAE, and 25 min, 134.6 °C, for the MAE system. The UAE system was identified as the most appropriate technique to obtain extracts with the highest amount of ergosterol, followed by MAE and HAE. Overall, these results demonstrate the potential of using the fruiting bodies of A. blazei as a suitable source to obtain ergosterol-rich extracts, which can be used as an ingredient for various biobased industrial applications. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019); L. Barros thanks national funding by FCT, P.I., through the individual scientific employment program-contract for her contract; to the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project MobilizadorNorte-01- 0247-FEDER-024479: ValorNatural®; to European Structural and Investment Funds (FEDER), through the Rural Development Program (PDR2020), within the scope of Project MicoCoating (PDR2020-101- 031472). The authors thank the GAIN (Xunta de Galicia) for financial support (P.P. 0000 421S 140.08) to M.A. Prieto by a post-doctoral (modality B) grant. R.C.G. Correa thanks CAPES Foundation, Ministry of Education, Brazil (CAPES fellow, process number 88881.120010/ 2016–01), for the financial support provided for her postdoctoral research in Polytechnic Institute of Bragança. Rosane Marina Peralta and Adelar Bracht are recipients of scientific productivity research grants from CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico, Brazil), grants numbers 307944/2015-8 and 304090/ 2016-6, respectively. To project POCI-01-0145-FEDER-006984 – Associate Laboratory LSRE-LCM funded by FEDER through COMPETE2020-POCI and by FCT. info:eu-repo/semantics/publishedVersion

The aim of the present work was to investigate, in a more extensive way, the oxidative state and parameters related to energy metabolism of the heart tissue of rats using the model of adjuvant-induced arthritis. The latter is a model for the human arthritic disease. Measurements were done in the total tissue homogenate, isolated mitochondria and cytosolic fraction. The adjuvant-induced arthritis caused several modifications in the oxidative state of the heart which, in general, indicate an increased oxidative stress (+80% reactive oxygen species), protein damage (+53% protein carbonyls) and lipid damage (+63% peroxidation) in the whole tissue. The distribution of these changes over the various cell compartments was frequently unequal. For example, protein carbonyls were increased in the whole tissue and in the cytosol, but not in the mitochondria. No changes in GSH content of the whole tissue were found, but it was increased in the mitochondria (+33%) and decreased in the cytosol (-19%). The activity of succinate dehydrogenase was 77% stimulated by arthritis; the activities of glutamate dehydrogenase, isocitrate dehydrogenase and cytochrome c oxidase were diminished by 31, 25 and 35.3%, respectively. In spite of these alterations, no changes in the mitochondrial respiratory activity and in the efficiency of energy transduction were found. It can be concluded that the adjuvant-induced arthritis in rats causes oxidative damage to the heart with an unequal intracellular distribution. Compared to the liver and brain the modifications caused by arthritis in the heart are less pronounced on variables such as GSH levels and protein integrity. Possibly this occurs because the antioxidant system of the heart is less impaired by arthritis than that reported for the former tissues. Even so, the modifications caused by arthritis represent an imbalanced situation that probably contributes to the cardiac symptoms of the arthritis disease.

This work compares the phenolic contents and the total antioxidant capacity of the 36 most popular Brazilian foods submitted to aqueous extraction or in vitro digestion. The purpose was to evaluate the extent by which digestion differs from the simple aqueous extraction procedures of several food matrices. After in vitro digestion, cereals, legumes, vegetables, tuberous vegetables, chocolates and fruits showed higher phenolic contents and higher antioxidant activities than those obtained by aqueous extraction. Contrarily, the digestion caused a reduction in the phenolic contents and antioxidant activities of beverages (red wine, coffee and yerba mate). Our results suggest that the phenolics of food groups with solid and complex matrix are protected against enzymatic action and alteration in pH during the digestion, what does not occur in liquid food matrices such as the beverages. This fact would overestimate the antioxidant activities of beverages submitted solely to aqueous extraction.

Citrus aurantium (bitter orange) extracts have been used in products for weight management and sports performance. These extracts contain large amounts of p-synephrine and much smaller amounts of p-octopamine. Both protoalkaloids exert lipolytic and glycogenolytic activities at similar concentrations. The biotransformation of p-synephrine and p-octopamine is not as well-known as those of other adrenergic amines. For this reason transformation of these amines was investigated in the isolated perfused liver. Special attention was devoted to the single pass extraction of each compound as well as to the kinetics of uptake. The assay of the amines in the outflowing perfusate was done by means of high performance liquid chromatography (HPLC). The single pass extraction of p-synephrine was higher than 90% at a portal concentration of 10 μM. It declined with the concentration, but was still around 30% at the concentration of 500 μM. At low concentrations (10-50 μM) the decreasing sequence of single pass extractions was p-synephrine > p-octopamine ≈ epinephrine > norepinephrine. Rates of uptake versus p-synephrine concentration resulted in a Michaelis-Menten type of relationship, with a KM value of 290.7 ± 32.1 μM and a Vmax of 0.762 ± 0.042 μmol min(-1) g(-1). The rates of uptake of p-octopamine did not present clear saturation and could be approximated by a linear relationship with a first order rate constant of 1.5 min(-1). The rapid hepatic transformation of p-synephrine and p-octopamine means that their concentration in the portal vein exceeds that in the systemic circulation during absorption. Their metabolic effects will, thus, be exerted predominantly in the liver.

Leaf and root extracts of Smallanthus sonchifolius (yacon), have antihyper-glycemic activity and antioxidant properties. The present study aims to compare the in vivo hepatic antioxidant activity of hydroalcoholic extracts of yacon leaves and roots in rats with streptozotocin-induced diabetes in terms of their in vitro antioxidant capacity. Rats were treated during 14 days with 1060 mg·Kg-1 root extract or 400 mg·Kg-1 leaf extract. The latter was richer in phenolics and possessed a much higher in vitro antioxidant activity. Both extracts prevented hyperglycemia in diabetic rats. The liver of diabetic rats presented increased levels of protein carbonyls and ROS and decreased activities of antioxidant enzymes. Treatment with both root and leaf extracts restored the protein carbonyl levels to normality. The root extract also restored the ROS levels to normality, but the leaf extract was not effective. The root extract was also more effective in restoring the activity of at least two important antioxidant enzymes (glucose 6-phosphate dehydrogenase and glutathione peroxidase). In terms of the antioxidant load (which was 17 times lower in the root extract treatment), the in vivo action of the root extract was more effective than the leaf extract in reducing the hepatic oxidative stress that accompanies diabetes.

Amylases are interesting targets for antidiabetic drugs because their inhibition is able to lower glycaemia without the need of hormonal control, as promoted by insulin or glibenclamide. In this context, the comparison between the binding features of α-amylases with their substrate and known inhibitors may provide insights aiming at the discovery of new antidiabetic drugs. In this work, the structure of the porcine pancreatic α-amylase was modelled with the acarbose pentasaccharide inhibitor, and used in structure-based virtual screening simulations based on a library containing the structures of amylose (AMY), acarbose (ACA) and the more representative structures of condensed tannin (CTN) and hydrolysable tannin (HTN). After validation of the methodology by redocking (mean rmsd ~ 0.8 A), the scores provided by programs AutoDock/Molegro were contradictory (− 1.5/− 23.3; − 3.5/− 24.6; − 4.3/− 14.6; −/− 19.5 for AMY, ACA, CTN and HTN respectively), indicating that a more sensitive methodology was necessary. The ΔGbinding was calculated by the molecular mechanics Poisson–Boltzmann surface area (MM-PBSA) method, which indicated that the HTN, ACA and CTN had higher affinities for the enzyme regarding the AMY substrate, with values of − 350.0, − 346.2, − 320.5 and − 209.2 kJ mol−1, respectively. The predicted relative affinities of HTN and CTN are in agreement with those obtained experimentally. The results provided useful information for the characterization of tannin binding to α-amylase, which can be applied in future studies aiming at finding new hypoglycaemic molecules among natural products.

Gluconeogenesis overstimulation due to hepatic insulin resistance is the best-known mechanism behind elevated glycemia in obese subjects with hepatic steatosis. This suggests that glucose production in fatty livers may differ from that of healthy livers, also in response to other gluconeogenic determinant factors, such as the type of substrate and modulators. Thus, the aim of this study was to investigate the effects of these factors on hepatic gluconeogenesis in cafeteria diet-induced obese adult rats submitted to a cafeteria diet at a young age. The livers of the cafeteria group exhibited higher gluconeogenesis rates when glycerol was the substrate, but lower rates were found when lactate and pyruvate were the substrates. Stearate or glucagon caused higher stimulations in gluconeogenesis in cafeteria group livers, irrespective of the gluconeogenic substrates. An increased mitochondrial NADH/NAD+ ratio and a reduced rate of 14CO2 production from [14C] fatty acids suggested restriction of the citric acid cycle. The higher glycogen and lipid levels were possibly the cause for the reduced cellular and vascular spaces found in cafeteria group livers, likely contributing to oxygen consumption restriction. In conclusion, specific substrates and gluconeogenic modulators contribute to a higher stimulation of gluconeogenesis in livers from the cafeteria group.

Methyl jasmonate (MeJA) is a fatty acid-derived cyclopentanone which shares structural similarities with prostaglandins and has been under study as a promising anti-inflammatory agent. This study investigated the actions of MeJA on systemic inflammation and oxidative status in rats with adjuvant-induced arthritis, a model for rheumatoid arthritis. MeJA (75 to 300 mg·kg−1) was administrated orally during 18 days after arthritis induction with Freund’s adjuvant. Articular and systemic inflammation was greatly increased in arthritic rats, likewise the oxidative stress in plasma and liver. The hepatic glucokinase activity and glycolysis were increased in arthritic rats. MeJA decreased most inflammatory parameters and abolished the increased protein carbonylation in plasma and liver, diminished the increased hepatic ROS content, and restored the hepatic GSH/GSSG ratio in arthritic rats. However, the MeJA treatment decreased the hepatic glucokinase activity and glycolysis and stimulated mitochondrial ROS production in healthy and arthritic rats. Oxygen uptake was increased by MeJA only in livers from treated arthritic rats. This action may bear relation to the increased activity of mitochondrial NADP+-dependent enzymes to provide reducing equivalents for the glutathione cycle. These beneficial effects, however, are associated with a decreased glucose flux through the glycolysis in the liver of arthritic and healthy rats.

Severe rheumatoid cachexia is associated with pronounced loss of muscle and fat mass in patients with advanced rheumatoid arthritis. This condition is associated with dyslipidemia and predisposition to cardiovascular diseases. Circulating levels of triglycerides (TG) and free fatty acids (FFA) have not yet been consistently defined in severe arthritis. Similarly, the metabolism of these lipids in the arthritic liver has not yet been clarified. Aiming at filling these gaps this study presents a characterization of the circulating lipid profile and of the fatty acids uptake and metabolism in perfused livers of rats with adjuvant-induced arthritis. The levels of TG and total cholesterol were reduced in both serum (10-20%) and liver (20-35%) of arthritic rats. The levels of circulating FFA were 40% higher in arthritic rats, possibly in consequence of cytokine-induced adipose tissue lipolysis. Hepatic uptake and oxidation of palmitic and oleic acids was higher in arthritic livers. The phenomenon results possibly from a more oxidized state of the arthritic liver. Indeed, NADPH/NADP+ and NADH/NAD+ ratios were 30% lower in arthritic livers, which additionally presented higher activities of the citric acid cycle driven by both endogenous and exogenous FFA. The lower levels of circulating and hepatic TG possibly are caused by an increased oxidation associated to a reduced synthesis of fatty acids in arthritic livers. These results reveal that the lipid hepatic metabolism in arthritic rats presents a strong catabolic tendency, a condition that should contribute to the marked cachexia described for arthritic rats and possibly for the severe rheumatoid arthritis.

Anti-inflammatory and antioxidant properties have been attributed to constituents of Rosmarinus officinalis. Considering the inflammatory nature of arthritis and the oxidative stress induced by the disease, this work aimed to investigate if R. officinalis is able to attenuate oxidative and inflammatory injuries caused by the disease. Rats with complete Freund's adjuvant induced arthritis were used. An aqueous extract of the rosemary leaves was administered orally. The parameters related to oxidative stress were measured in the liver, brain and plasma. The administration of the rosemary extract (RE) to arthritic rats diminished oxidative damage (e.g., less carbonylated proteins), improved the oxidative state (e.g., less reactive oxygen species) and also increased the antioxidant capacity by increasing the GSH level and the GSH/GSSG ratio and by almost normalizing the activity of several antioxidant enzymes. The treatment of arthritic rats with the extract also diminished paw edema, the number of leukocytes recruited in the femoro-tibial joint cavities and the weight of the lymph nodes and delayed the appearance of secondary lesions. Twelve phenolic compounds were identified and quantified in the aqueous RE, rosmarinic acid being the most abundant one. In addition, the extract also contains polysaccharides and proteins. The anti-inflammatory and antioxidant activities of the RE can be attributed, partly at least, to its content of polyphenolics with the strong possibility of synergistic interactions as well as metabolic activations. The results corroborate and amplify the general notion that rosemary aqueous extracts possess efficient anti-inflammatory agents and suggest that they are able to attenuate the oxidative stress inherent to arthritis.

The current study investigated the action of β-caryophyllene, the major constituent of copaiba oil, on the systemic inflammation, oxidative status, and liver cell metabolism of rats with adjuvant-induced arthritis, a model for rheumatoid arthritis. This study also compared the actions of β-caryophyllene with those previously reported for copaiba oil on arthritic rats. For this purpose, Holtzman healthy and arthritic rats received 215 and 430 mg·kg-1 β-caryophyllene orally once a day during 18 days. Both doses of β-caryophyllene reduced the adjuvant-induced paw edema, swollen of lymph nodes, and number of circulating and articular leukocytes. β-Caryophyllene, at the dose of 430 mg·kg -1 , abolished the increases of protein carbonyl groups and myeloperoxidase activity in the liver and plasma of arthritic rats and, at both doses, it restored the increased levels of reactive oxygen species and reduced glutathione in the arthritic liver. These beneficial actions were of the same extension as those of copaiba oil ( Copaifera reticulata) and, therefore, β-caryophyllene is possibly responsible for the anti-inflammatory and antioxidant actions of the oil. Hepatic gluconeogenesis was 40% lower in arthritic rats, which also presented a reduced number of hepatocytes per liver area (-23%) associated with increased hepatocyte area (+18%) and liver weight (+50%). None of these hepatic alterations were improved by β-caryophyllene, but not even by ibuprofen. However, unlike copaiba oil, β-caryophyllene did not modify the hepatic morphology and metabolism of healthy rats. These results reveal that β-caryophyllene improves the systemic inflammation and oxidative status of arthritic rats and, in addition, it was not associated with hepatotoxicity.

In recent years, mycosterols have emerged as potential functional ingredients for the development of sterol-enriched food products and dietary supplements. Agaricus blazei is a mushroom rich in bioactive compounds. For commercial purposes, their fruiting bodies must obey rigid morphological criteria. Those not conforming to these criteria are usually discarded, although this does not mean impairment of their content in bioactives. The aim of the present work was to propose the use of commercially discarded A. blazei fruiting bodies for obtaining an extract rich in ergosterol as a fortifier ingredient for yogurts. For extraction, the Soxhlet technology was used and the highest ergosterol yield (around 12%) was achieved in the 5th cycle, yielding 58.53 ± 1.72 μg of ergosterol per 100 g of mushroom (dry weight). The ergosterol rich extract presented notable antioxidant and antimicrobial properties, besides showing no hepatotoxicity. When added to the yogurts it significantly enhanced their antioxidant properties. Furthermore, it did not significantly alter the nutritional or the individual fatty acid profiles of the final dairy products. Thus, A. blazei fruiting bodies that do not conform to the commercial requirements of the market and are normally discarded could be exploited for obtaining a natural high added-value food additive, following the circular bioeconomy concept. R. C. G. Correa thanks the CAPES Foundation, Ministry of Education, Brazil (CAPES fellow, process number 88881.120010/2016-01), for the financial support provided for her postdoctoral research in the Polytechnic Institute of Bragança. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and the FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/ 00690/2013), L. Barros contract and A. Fernandes grant (SFRH/ BPD/114753/2016). This work was also funded by the European Structural and Investment Funds (FEEI) through the Regional Operational Program North 2020, within the scope of Project Mobilizador ValorNatural®. Rosane Marina Peralta and Adelar Bracht are recipients of scientific productivity research grants from CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico, Brazil), grants numbers 307944/2015- 8 and 304090/2016-6, respectively. M. Sokovic is grateful for financial support to the Ministry of Education, Science and Technological Development of Republic of Serbia, Grant No. 173032. info:eu-repo/semantics/publishedVersion

South to Northeast of Brazil, ora-pro-nobis leaves are used in traditional cuisine, featuring as an ingredient of various sweet and savoury dishes. In some low-income communities, it is known as the ‘meat of the poor’, being the main protein source available.1 The aim of the present work was to perform an inedited study on the phytochemical profile and some biological activities of P. aculeate leaves, with the view of expanding the current knowledge on the potentialities of this superfood. For this purpose, its hydroethanolic extract was characterized in terms of phenolic composition, antioxidant, and antibacterial capabilities and finally the hepatotoxicity of the extract was evaluated. A total of ten phenolic compounds were identified via high-performance liquid chromatography coupled to diode array detector and mass spectrometer (HPLC–DAD–ESI/MSn): two phenolic acids (caffeic acid derivatives) and eight flavonoids (quercetin, kaempferol and isorhamnetin glycoside derivatives). Caftaric acid was the extract’s major phenolic constituent, accounting for more than 49% of the phenolic content, followed by quercetin-3-O-rutinoside (14.99%) and isorhamnetin-O-pentoside-O-rutinoside (9.56%). The total phenolic content found in the hydromethanolic extract was relevant (23.75 mg/g). A broad study of the extracts’ antioxidant activity was carried out, using a set of five distinct methods: two cell-based methods (1) oxidative haemolysis inhibition assay (OxHLIA); (2) the inhibition of the production of thiobarbituric acid reactive substances (TBARS); besides three chemical in vitro assays (3) reduction of the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH); (4) reduction of the 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonate) cation (ABTS) and (5) hydroxyl radical scavenging assay.2 Overall, the ora-pro-nobis leaf extract showed relevant values of antioxidant activity, even higher than Trolox in the DPPH and ABTS trials. The antioxidant activity verified for the hydroethanolic leaf extract of P. aculeata is possibly related to its phenolic composition; the predominant constituents of the extract, i.e. caftaric acid and rutin, have widely proven antioxidant activities.1 Furthermore, the evaluated extract showed no toxicity against a non-tumour liver primary culture PLP2, at the highest tested concentration (400 μg/mL). The antimicrobial activity exhibited by the extract against both Gram-positive (Enterococcus faecalis, Listeria monocytogenes, MRSA – Methicillin-resistant Staphylococcus aureus) and Gram-negative (Escherichia coli, Klebsiella pneumoniae, Morganella morganii, Proteus mirabilis, and Pseudomonas aeruginosa) bacteria (MIC values between 5 to 20 mg/mL) suggests the presence of a broad spectrum of phytochemicals with antimicrobial activity. The extract was more active against K. pneumonia than the antibiotic ampicillin, whereas against M. morganii it presented equivalent inhibitory effect. Therefore, the information reported here not only corroborates the importance of the production and consumption of P. aculeata leaves by the low income population to improve nutrition, but also reinforces their potential as a sustainable source of promising food ingredients to be used both for food enrichment and preservation. The authors are grateful to FCT - Portugal and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019), L. Barros, R. C. Calhelha and C. Pereira contracts (through national FCT funds, under the celebration of the contract program of institutional scientific employment). J. A. A. Garcia thanks Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). R.C.G. Corrêa (Process number 167378/2017-1), R.M. Peralta (Project number 307944/2015-8) and A. Bracht (Project number 304090/2016-6) thank Conselho Nacional de Desenvolvimento Científico e Tecnologia (CNPq), Brazil, for the financial support. Funding: To FEDER-Interreg España-Portugal for financial support of the project 0377_Iberphenol_6_E and TRANSCoLAB 0612_TRANS_CO_LAB_2_P; FEDER under the Regional North Operational Program 2020, to the Mobilizing Project Norte-01-0247-FEDER-024479: ValorNatural®. info:eu-repo/semantics/publishedVersion

The potential phytochemical losses occurring throughout the sequential steps of in-vitro gastrointestinal digestion and colonic fermentation of a rosemary aqueous extract were investigated. Crude (CE), digested (DE) and fermented (FE) extracts were characterized in terms of their phenolic profile and biological activities. Rosmarinic acid was the phytochemical that underwent the most significate transformation during digestion and fermentation, which amounted to 60% compared to the 26% degradation of the total phenolics. Overall, the simulated digestion step decreased the antioxidant activity estimated by DPPH, ABTS, FRAP, ORAC and TBARS assays. Both CE and DE did not present antiproliferative potential, however, FE exhibited a pronounced cytotoxic activity (GI50 = 116 µg/mL) against HeLa cells. CE and DE showed to be moderate inhibitors of methicillin-resistant Staphylococcus aureus (MRSA), methicillin-susceptible S. aureus (MSSA), S. aureus, Listeria monocytogenes, whilst the FE acted as a moderate inhibitor of MRSA and MSSA. G.A. Gonçalves and V.G. Correa thank Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for the financial support provided for their post-graduate studies in the State University of Maringá. R.C.G. Corrêa thanks Conselho Nacional de Desenvolvimento Científico e Tecnologia ( CNPq ) for financing her postdoctoral research at State University of Maringá (Process number 167378/2017-1). R.M. Peralta (Project number 307944/2015-8) and A. Bracht (Project number 304090/2016-6) are CNPq research grant recipients. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013), L. Barros and R. Calhelha contracts; and also thank to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E. Appendix A info:eu-repo/semantics/publishedVersion

The pulp of the fruits of Juçara (Euterpes edulis Martius), a native tree of the Atlantic Rainforest of Brazil, is widely consumed thanks to its flavour and nutritional value. The industrial production of Juçara fruit pulp generates solid residues (peel) which are usually discarded. In this work, a hydroalcoholic extract from Juçara peel flour was evaluated for its phenolic profile as well as for its bioactivities. A total of nineteen phenolic compounds were identified in the Juçara peel flour. Among these, seventeen were non-anthocyanin phenolic compounds, namely two phenolic acids, four flavanonols, six flavones, and five flavonols; whereas the two anthocyanin molecules were cyanidin glycoside derivatives. The Euterpe edulis peel flour presented antioxidant activity and antibacterial potential but was not hepatotoxic. These observations corroborate the idea that this byproduct could fit well into the circular bioeconomy concept, thus promoting the Juçara fruit production chain. J.A.A. Garcia thanks Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for the financial support provided for her masters studies in the State University of Maringá. R.C.G. Corrêa thanks Conselho Nacional de Desenvolvimento Científico e Tecnologia (CNPq) for financing her postdoctoral research at State University of Maringá (Process number 167378/2017-1). R.M. Peralta (Project number 307944/2015-8) and A. Bracht (Project number 304090/2016-6) are CNPq research grant recipients. The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/ 00690/2013), L. Barros and R. C. Calhelha contracts; to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E; the European Regional Development Fund (ERDF) to FEDER through the Regional Operational Program North 2020, within the scope of Project NORTE-01-0145-FEDER-023289: DeCodE and project Mobilizador Norte-01-0247-FEDER-024479: ValorNatural®. info:eu-repo/semantics/publishedVersion

Healthy and adjuvant-induced arthritic rats were treated for 23 days with daily doses of 400 and 800 mg kg−1 Ilex paraguariensis extract. This treatment (a) diminished the ROS levels in the liver and brain, (b) decreased oxidative protein and lipid damage in liver and brain, (c) increased the plasma antioxidant capacity, (d) increased the GSH levels and the GSH/GSSH ratio in both the liver and the brain, (e) almost restored the enzymatic activities linked to the metabolism of GSH–GSSG, and (f ) reversed the modified activities of xanthine oxidase, superoxide dismutase and catalase. The anti-inflammatory actions (firstly) and the antioxidant actions (in the second place) of the yerba mate constituents (e.g., chlorogenic acid derivatives) are the causes of these beneficial effects. Daily ingestion of traditional yerba mate beverages may be effective in attenuating the symptoms of inflammatory diseases, especially in older adults. This work was financially supported by grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-307944/2015-8), Coordenação do Aperfeiçoamento de Pessoal do Ensino Superior (CAPES) and Fundação Araucária. The authors are also indebted to Jailson Araújo Dantas for his technical assistance and to Dr Ciomar A. B. Amado for facilitating access to equipment of the Pharmacology and Therapeutics Department of the State University of Maringá. The authors are also grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013) and L. Barros contract. info:eu-repo/semantics/publishedVersion

Background: Non-digestible oligosaccharides are versatile sources of chemical diversity, well known for their prebiotic actions, found naturally in plants or produced by chemical or enzymatic synthesis or by hydrolysis of polysaccharides. Compared to polyphenols or even polysaccharides, the antioxidant potential of oligosaccharides is still unexplored. The aim of the present work was to provide an up-to-date, broad and critical contribution on the topic of antioxidant oligosaccharides. Methods: The search was performed by crossing the words oligosaccharides and antioxidant. Whenever possible, attempts at establishing correlations between chemical structure and antioxidant activity were undertaken. Results: The most representative in vitro and in vivo studies were compiled in two tables. Chitooligosaccharides and xylooligosaccharides and their derivatives were the most studied up to now. The antioxidant activities of oligosaccharides depend on the degree of polymerization and the method used for depolymerization. Other factors influencing the antioxidant strength are solubility, monosaccharide composition, the type of glycosidic linkages of the side chains, molecular weight, reducing sugar content, the presence of phenolic groups such as ferulic acid, and the presence of uronic acid, among others. Modification of the antioxidant capacity of oligosaccharides has been achieved by adding diverse organic groups to their structures, thus increasing also the spectrum of potentially useful molecules. Conclusion: A great amount of high-quality evidence has been accumulating during the last decade in support of a meaningful antioxidant activity of oligosaccharides and derivatives. Ingestion of antioxidant oligosaccharides can be visualized as beneficial to human and animal health.

This study investigated the antioxidant, antimicrobial and cytotoxic properties of a purified yerba mate polysaccharide. The yerba mate polysaccharide showed a prominent antioxidant activity as evaluated by 2,2-diphenyl-1- picrylhydrazyl (DPPH•)-radical scavenging activity (IC50 = 1.25 ± 0.10 mg/mL), 3-ethyl benzothiazoline-6- sulphonic acid (ABTS•+)-radical scavenging activity (IC50 = 0.41 ± 0.05 mg/mL), and hydroxyl scavenging activity (IC50= 3.36 ± 0.31 mg/mL). The antioxidant activity evaluated as the ferric ion reduction power (FRAP) and oxygen radical absorbance radical assay (ORAC), expressed as trolox equivalents,were 20.84±1.61 μMTE/- mg and 556.30± 12.83 μM TE/mg, respectively. The purified yerbamate polysaccharide presented high antimicrobial activity against several bacterial and fungal strains; however, no cytotoxicity against all four tumor human cell lines assessed. The authors thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Proc. 3079/2015-8) for funding this study. Author V.G. Correa thanks Coordenação de Aperfeiçoamento do Pessoal do Ensino Superior (CAPES) for the financial support provided for their post-graduate studies in Universidade Estadual de Maringá. R.C.G. Corrêa thanks CAPES Foundation,Ministry of Education, Brazil (process number 88881.120010/2016–01) for funding her postdoctoral internship in Polytechnic Institute of Bragança. A. Bracht, R.A. Peralta and R.M. Peralta are research grant recipients of CNPq. info:eu-repo/semantics/publishedVersion