Enhancing Vascular Health and Lowering Blood Pressure in Spontaneously Hypertensive Rats through Syrah Grape (Vitis vinifera) Pomace: The Role of Phenolic Compounds.

BACKGROUND: The beneficial properties of wine by-products include actions that help prevent and treat cardiovascular conditions such as hypertension, primarily due to their antioxidant effects. Novel pharmacotherapies are being developed to treat arterial hypertension, including investigations into natural products exhibiting biological activity, necessitating rigorous evaluation of their efficacy and safety. This study aimed to identify and quantify phenolic compounds in Syrah (Vitis vinifera) grapes grown in the Brazilian Cerrado and their presence in winemaking by-products. It also examined the effects of grape pomace on blood pressure. METHODS: Fresh grapes, pomace, and lees, were subjected to spectrophotometric determination of total phenolic compounds, followed by identification and quantification using HPLC-DAD-ESI-MSn. Normotensive male rats (Wistar) and spontaneously hypertensive rats (SHR) received grape pomace-enriched (150 or 300 mg/kg/day, 14 days) or standard chow. Indirect arterial pressure was assessed, while vascular reactivity was evaluated in mesenteric resistance arteries. RESULTS: Pomace samples exhibited higher total phenolic compound concentrations than grapes or lees. Seven derivatives of hydroxycinnamic acids and twenty-one flavonols were identified. Quercetin-3-glucoside and ethyl caffeate were the most abundant phenolic compounds. Grape pomace-enriched chow demonstrated a dose-dependent hypotensive effect in rats. CONCLUSION: the abundance of flavonols and hydroxycinnamic acids, combined with their hypotensive effects, underscores the therapeutic potential of fine wine-making by-products produced in the Brazilian Cerrado.

A maize enzyme from the 2-oxoglutarate-dependent oxygenase family with unique kinetic properties, mediates resistance against pathogens and regulates senescence.

In plants, salicylic acid (SA) hydroxylation regulates SA homoeostasis, playing an essential role during plant development and response to pathogens. This reaction is catalysed by SA hydroxylase enzymes, which hydroxylate SA producing 2,3-dihydroxybenzoic acid (2,3-DHBA) and/or 2,5-dihydroxybenzoic acid (2,5-DHBA). Several SA hydroxylases have recently been identified and characterised from different plant species, but no such activity has yet been reported in maize. In this work, we describe the identification and characterisation of a new SA hydroxylase in maize plants. This enzyme, with high sequence similarity to previously described SA hydroxylases from Arabidopsis and rice, converts SA into 2,5-DHBA; however, it has different kinetic properties to those of previously characterised enzymes, and it also catalysers the conversion of the flavonoid dihydroquercetin into quercetin in in vitro activity assays, suggesting that the maize enzyme may have different roles in vivo to those previously reported from other species. Despite this, ZmS5H can complement the pathogen resistance and the early senescence phenotypes of Arabidopsis s3h mutant plants. Finally, we characterised a maize mutant in the S5H gene (s5hMu) that has altered growth, senescence and increased resistance against Colletotrichum graminicola infection, showing not only alterations in SA and 2,5-DHBA but also in flavonol levels. Together, the results presented here provide evidence that SA hydroxylases in different plant species have evolved to show differences in catalytic properties that may be important to fine tune SA levels and other phenolic compounds such as flavonols, to regulate different aspects of plant development and pathogen defence.

Characterisation of bioactive compounds from leaves of Smilax fluminensis Steud. by gc-MS and LC-DAD-MS, and analysis of its antioxidant and ecotoxicological activities.

Smilax fluminensis Steud. is distributed in Central and South America, commonly named as 'salsaparilha' or 'japicanga'. In the present work, the chemical composition was determined, antioxidant and cytotoxic effects were evaluated for the ethanol extract (EE) and fractions from leaves. Fatty acid esters and phytol were characterised in the hexane (HEX) fraction. O-glycosylated flavonols and flavones, caffeic acid derivatives, and steroidal saponins were annotated for EE, and dichloromethane (DCM), ethyl acetate (AC), and hydroethanol (HE) fractions. The samples contain flavonoids and phenolic compounds, and the AC fraction displayed the biggest concentration of these substances. Antioxidant potential was observed in all samples, being especially pronounced in the AC fraction by DPPH and FRAP assays, with IC50 values of 8.18 and 2.35 µg/mL, respectively. AC and HEX fractions showed 35% and 5% lethality at 1000 µg/mL, in the Artemia salina assay, and the other samples did not show a toxic effect.

Antioxidant and Antifungal Activities and Characterization of Phenolic Compounds Using High-Performance Liquid Chromatography and Mass Spectrometry (HPLC-MS) in Empetrum rubrum Vahl ex Willd.

In searching for compounds with antioxidant and antifungal activity, our study focused on the subshrub species Empetrum rubrum Vahl ex Willd. (Ericaceae). We measured the antioxidant activity of its methanolic extract (MEE) obtained from the aerial parts (leaves and stems) and of its methanolic extract (MEF) obtained from the lyophilized fruits. The antioxidant activity of the MEE and MEF was evaluated in vitro via a 2,2-Diphenyl-1-picrylhydrazyl (DPPH) free radical and 2,2'-Azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) cationic radical. The results were expressed in gallic acid and Trolox equivalents for the DPPH and ABTS assays, respectively. The antioxidant activities, for the DPPH and ABTS assays, were also evaluated by considering the IC50 values. Concerning the antioxidant activity, the total phenolic content (TPC) in the MEE and MEF was determined using the Folin-Ciocalteu method. Polyphenols contained in the leaves, stems, and fruits of E. rubrum were determined qualitatively by employing high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) analysis. The antifungal activity of the MEE obtained from the aerial parts of E. rubrum was tested against Rhizoctonia solani. The results of IC50 values measured by the DPPH and ABTS methods with MEE were 0.4145 ± 0.0068 mg mL-1 and 0.1088 ± 0.0023 mg mL-1, respectively, and the IC50 values for MEF were 6.4768 ± 0.0218 mg mL-1 and 0.7666 ± 0.0089 mg mL-1 measured by the DPPH and ABTS methods, respectively. The HPLC-MS analysis revealed the presence of anthocyanins, phenolic acids derivatives, and flavonols. In vitro, mycelial growth of this fungus was reduced from 90% to nearly 100% in the presence of MEE. The observed antifungal effect is related to the presence of the abovementioned phenols, detected in the MEE.

Gall inducer Dasineura sp. alters the polyphenol profile and antioxidant activity of Peumus boldus stems.

Peumus boldus, a tree native to Chile, is extensively used for medicinal purposes due to its richness in alkaloids and antioxidant polyphenols. A species of galling insect, Dasineura sp. induces structural and chemical changes on P. boldus stems while its galls are established and developed. Taking into account the antioxidant properties of P. boldus polyphenols, it would be expected that Dasineura sp. induces changes in the accumulation sites, chemical profile, and antioxidant activity of the P. boldus stem polyphenols, related to different reactive oxygen species (ROS) production levels during gall development. Dasineura sp. induces changes in the accumulation sites of total polyphenols, flavonols, and lignin, redirecting their accumulation toward the sites of greatest production of H2O2 and O2.-. Although changes in total polyphenol content would be expected, this did not vary significantly between non-galled and galled stems. However, the galling insect induced changes in the profile and concentration of soluble polyphenols, leading to the gall extracts' antioxidant capacity decreasing significantly during the maturation and senescence stages. Additionally, during the maturation stage, lignin deposition increases in the more peripheral gall tissues, which also contributes to ROS dissipation. The differences in the different gall developmental stages' antioxidant activity could be related to the identity and concentration of phenolic compounds in each gall extract, rather than to the total phenol content. Regardless of the mechanisms involved, the dissipation of the ROS generated by Dasineura sp. activity occurs, restoring the redox balance in galls and guaranteeing the success of the inducer.

Anti-hyperglycemic potential and chemical constituents of Aristolochia triangularis Cham. leaves - A medicinal species native to Brazilian forests.

ETHNOPHARMACOLOGICAL RELEVANCE: Aristolochia triangularis Cham. has been used in Brazilian traditional medicine for various therapeutic purposes, including as a leaf-based infusion for diabetes management. AIM OF THE STUDY: This study was designed to chemically characterize an infusion of in natura A. triangularis leaves and evaluate the in vivo anti-hyperglycemic properties of this infusion. MATERIALS AND METHODS: Chemical composition was examined using liquid-liquid extraction procedure, chromatographic methods, NMR, and LC-MS/MS. The in vivo anti-hyperglycemic activity of the freeze-dried infusion of A. triangularis leaves (Inf-L-At) was assessed using oral glucose tolerance test (OGTT). Initially, normoglycemic male rats were pre-treated with orally administered Inf-L-At at doses of 62.5, 125, and 250 mg/kg for two consecutive days. On the day of the OGTT, fasting animals received a glucose load (4 g/kg) 30 min after treatment with Inf-L-At, and the blood glucose levels were verified at 15, 30, 60, and 180 min. Intestinal maltase, lactase, and sucrase activities and muscle and liver glycogen contents were also assessed after the OGTT. RESULTS: Inf-L-At extract led to glycemic reduction with no dose-response at 15, 30, and 60 min comparable to that of the antidiabetic drug glibenclamide and was accompanied by an increase in hepatic and muscle glycogen contents. Additionally, there was a significant statistically decrease in the in vitro activity of disaccharidases. Maltase and sucrase activities were inhibited at all doses, whereas lactase activity was inhibited only at 62.5 and 250 mg/kg. In total, 75 compounds were found in the infusion, including seven new ones, (7S*,8S*,7ꞌS*,8ꞌR*)-4,4ꞌ-dihydroxy-3,3ꞌ-dimethoxy-7,9ꞌ-epoxylignan-7ꞌ-ol; 4ꞌ-hydroxy-3ꞌ-methoxy-3,4-methylenedioxy-7,9ꞌ-epoxylignan-9,7ꞌ-diol; triangularisines A, B, and C; N-ethyl-N-methyl-affineine; and N-methyl pachyconfine, and one previously not described as a natural product, epi-secoisolariciresinol monomethyl ether. CONCLUSION: The results demonstrated the anti-hyperglycemic activity of the infusion from A. triangularis leaves and showed that it is a rich source of lignoids, alkaloids, and glycosylated flavonoids, which are known to exhibit antidiabetic effects and other biological properties that can be beneficial for patients with chronic hyperglycemia, thus certifying the popular use of this herbal drink.

Total Polyphenol Intake, Polyphenol Subtypes, and Prevalence of Hypertension in the CUME Cohort.

Objective:Epidemiological studies have shown associations between polyphenol consumption and reduced risk of cardiovascular diseases. This study aimed to assess the association between polyphenol intake and the prevalence of hypertension.Methods:This cross-sectional study was performed on data from the Cohort of Universities of Minas Gerais (CUME) project. Participants completed an online food frequency questionnaire, and polyphenol intake was assessed using the Phenol-Explorer database and articles. Hypertension was determined by a medical diagnosis, having a blood pressure ≥ 130 mmHg/80 mmHg, or using antihypertensive drugs. Adjusted logistic regression models were used to estimate the odds ratios (ORs) and 95% confidence intervals (95% CIs) for the prevalence of hypertension.Results:The prevalence of hypertension was 39.57%, and the average intake of total polyphenols was 860.79 mg/day. The highest (5th quintile) intake of flavonoids (mean: 368.46mg/day; OR: 0.83; 95%CI 0.70; 0.97), hydroxybenzoic acids (mean: 379.38mg/day; OR: 0.77; 95%CI: 0.66;0.91), and flavonols (mean: 44.13mg/day; OR: 0.79; 95%CI: 0.67; 0.93) was inversely associated with hypertension prevalence, compared to the lowest intake (1st quintile).Conclusions:Our findings demonstrate that the intake of flavonoids, hydroxybenzoic acids, and flavonols is associated with a lower prevalence of hypertension.

Phenolic composition of jussara (Euterpe edulisMartius) from Minas Gerais and Espírito Santo, Brazil, determined by high performance liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI-MS) / Composição fenólica de Jussara (Euterpe edulisMartius) de Minas Gerais e Espírito Santo, Brasil, determinada por cromatografia líquida de alta eficiência acoplada à detecção de arranjo de diodos e espectrometria de massa contendo sistema de ionização por eletrospray (HPLC-DAD-ESI MS/MS)

ABSTRACT: This research assessed the phenolic composition of Jussara pulp from the Brazilian states of Minas Gerais (MG) and Espírito Santo (ES) using HPLC-DAD-MS/MS. Seventeen anthocyanins were detected in fruits, derived from cyanidin, pelargonidin and peonidin. Among the non-anthocyanic phenolic compounds, flavonols (kaempferol, quercetin and isorhamnetin derivatives), flavan-3-ols (catechin, epicatechin, B-type procyanidins and unknown dimers) and resveratrol in its glycosylated form have been identified. Catechin (32.41-60.56 mg 100g-1) and epicatechin (18.86-40.92 mg 100g-1) were the main flavan-3-ols present in the fruits. The samples showed small concentrations of resveratrol glycosides (0.02-0.91 mg 100g-1). The analytical methodology used (HPLC-DAD-ESI-MS/MS) permitted the identification of newly reported compounds in this fruit.

RESUMO: O objetivo desta pesquisa foi avaliar a composição fenólica da polpa de Jussara dos Estados brasileiros de Minas Gerais (MG) e Espírito Santo (ES) usando HPLC-DAD-MS / MS. Dezessete antocianinas foram detectadas, dentre elas, derivadas de cianidina, pelargonidina e peonidina. Dentre os compostos fenólicos não antociânicos, foram identificados flavonóis (derivados de caempferol, quercetina e isorhamnetina), flavan-3-ols (catequina, epicatequina, procianidinas do tipo B e dímeros desconhecidos) e resveratrol em sua forma glicosilada. Catequina (32,41-60,56 mg 100g-1) e epicatequina (18,86-40,92 mg 100g-1) foram os principais flavan-3-óis presentes nas frutas. As amostras apresentaram pequenas concentrações de glicosídeos resveratrol (0,02-0,91 mg 100g-1). A metodologia analítica utilizada (HPLC-DAD-ESI-MS / MS) permitiu a identificação de novos compostos relatados presentes na composição da polpa de Jussara.

Chemical Analysis of Eruca sativa Ethanolic Extract and Its Effects on Hyperuricaemia.

In vivo assays and chemical analyses were performed on the ethanolic extract from leaves of Eruca sativa. UHPLC-ESI-QTOF analysis confirmed the presence of glucosinolates and flavonol glucosides. The major flavonoid of the ethanolic extract, kaempferol-3,4'-di-O-ß-glucoside, was isolated, a HPLC-DAD method developed and validated to quantify its content in the extract. In vivo experiments were carried out on Wistar rats with hyperuricaemia induced by potassium oxonate and uric acid. A hypouricaemic effect was observed in hyperuricaemic Wistar rats treated with ethanolic extract at dose of 125 mg/kg and kaempferol-3,4'-di-O-ß-glucoside at dose of 10 mg/kg. The main anti-hyperuricaemic mechanism observed in the extract was uricosuric. Kaempferol-3,4'-di-O-ß-glucoside was identified as an important component responsible for the total activity of the ethanolic extract and was considered as a good chemical and biological marker of the ethanolic extract of E. sativa. The obtained results indicated the potential of E. sativa in the treatment of hyperuricaemia and its comorbidities.

Antitumor effect of isoquercetin on tissue vasohibin expression and colon cancer vasculature.

Tumor cells trigger angiogenesis through the expression of angiogenic factors. Vasohibins (VASHs) are a family of peptides that regulate angiogenesis. Flavonoids have antiproliferative antitumor properties; however, few studies have highlighted their antiangiogenic potential. This study evaluated the flavonoid isoquercetin (Q3G) as an antitumor compound related to colon cancer vascularization and regulation of VASH1 and 2. Mice bearing xenogeneic colon cancer (n = 15) were divided into 3 groups: Q3G-treated (gavage, daily over a week), bevacizumab-treated (intraperitoneal, single dose), or untreated animals. Tumor growth, histological characteristics, blood vessel volume, and VASH1 and 2 expressions were analyzed. Q3G impaired tumor growth and vascularization, upregulated VASH1, and downregulated VASH2 in comparison to untreated animals. Mice treated with Q3G showed approximately 65% fewer blood vessels than untreated animals and 50% fewer blood vessels than mice treated with bevacizumab. Thus, we show that Q3G has antitumor activity, impairs vascularization, and differentially modulates VASH1 and 2 expressions in colon cancer.

Effects of flavonols on emotional behavior and compounds of the serotonergic system: A preclinical systematic review.

Serotonin (5-hydroxytryptamine, 5-HT) is a neurotransmitter that regulates multiple psychophysiological functions. An imbalance of 5-HT in the brain can modulate emotional behavior such as depression and anxiety. Substances, such as flavonols, naturally found in some plants and foods have beneficial effects on psychiatric disorders, have been studied. The aim of this systematic review was to investigate the effects of flavonols on morphological, physiological, and cellular aspects of the serotonergic system as well as on some behaviors modulated by this system. Literature searches were performed in the LILACS, Web of Science, Scopus, PubMed and Sigle via Open Grey databases, from which 1725 studies were found. Using a predefined protocol registered on the CAMARADES website, 18 studies were chosen for qualitative synthesis. Internal validity was assessed using the SYRCLE's risk of bias tool. The Kappa index was also measured to analyze agreement among the reviewers. The results of this systematic review showed that flavonols have been reported to modify physiological aspects of the serotonergic system, increasing levels of serotonin and decreasing levels of its metabolite, 5-hydroxyindoleacetic acid (5-HIAA) and promoting antioxidant effects in encephalic regions. Moreover, the results showed that flavonols can also modulate of the serotonergic system, being associated with antidepressant and anxiolytic activities. Additionally, flavonols were found to not have psychostimulant effect; they can, however, reverse damage to locomotor activity.

Analysis of the Molecular Mechanisms by Flavonoids with Potential Use for Osteoporosis Prevention or Therapy.

BACKGROUND: Osteoporosis is the most common skeletal disorder worldwide. Flavonoids have the potential to alleviate bone alterations in osteoporotic patients with the advantage of being safer and less expensive than conventional therapies. OBJECTIVE: The main objective is to analyze the molecular mechanisms triggered in bone by different subclasses of flavonoids. In addition, this review provides an up-to-date overview of the cellular and molecular aspects of osteoporotic bones versus healthy bones, and a brief description of some epidemiological studies indicating that flavonoids could be useful for osteoporosis treatment. METHODS: The PubMed database was searched in 2001- 2021 using the keywords osteoporosis, flavonoids, and their subclasses such as flavones, flavonols, flavanols, isoflavones, flavanones and anthocyanins, focusing the data on the molecular mechanisms triggered in bone. RESULTS: Although flavonoids comprise many compounds that differ in structure, their effects on bone loss in postmenopausal women or in ovariectomized-induced osteoporotic animals are quite similar. Most of them increase bone mineral density and bone strength, which occur through an enhancement of osteoblastogenesis and osteoclast apoptosis, a decrease in osteoclastogenesis, as well as an increase in neovascularization on the site of the osteoporotic fracture. CONCLUSION: Several molecules of signaling pathways are involved in the effect of flavonoids on osteoporotic bone. Whether all flavonoids have a common mechanism or they act as ligands of estrogen receptors remains to be established. More clinical trials are necessary to know better their safety, efficacy, delivery and bioavailability in humans, as well as comparative studies with conventional therapies.

High antioxidant activity of phenolic compounds dampens oxidative stress in Espinosa nothofagi galls induced on Nothofagus obliqua buds.

Reactive oxygen species (ROS) are considered the first signaling molecules involved in gall development, linked to the establishment of cyto-histological gradients leading to gall tissue redifferentiation. ROS overproduction induces the failure of gall establishment or its premature senescence. Galls could therefore have efficient mechanisms of ROS dissipation and maintenance of homeostasis, such as polyphenol synthesis. The co-occurrence of ROS and polyphenols in the Espinosa nothofagi galls induced on Nothofagus obliqua buds was explored and was related to the antioxidant capacity of the inner (IC) and outer (OC) gall compartments. We hypothesize that: (i) ROS are produced and accumulated in both tissue compartments of E. nothofagi galls in co-occurrence with polyphenolic, flavonols, and lignin, conferring high antioxidant activity to inner and outer gall tissue compartment; (ii) antioxidant activity is higher in IC related to a higher polyphenol concentration in this compartment. The results show that ROS and polyphenols, mainly flavonols, are produced and accumulated in IC and OC, while lignin accumulated mainly in the IC. In both gall compartments, polyphenols mediate ROS elimination, confirmed by histochemical and spectrophotometry techniques. The IC extract has the highest antioxidant capacity, probably due to lignin deposition and a higher polyphenol concentration in this compartment.

Ação antimicrobiana e antibiofilme de flavonóis isolados ou incorporados em hidrogéis termossensíveis e sua influência sobre marcadores de mineralização em células odontoblastóides para aplicação em endodontia / Antimicrobial/antibiofilm action of flavonols alone or incorporated in thermosensitive hydrogels and their influence on mineralization markers in odontoblast-like cells for endodontic applications

Ainda não foi encontrado um medicamento capaz de desinfetar os canais radiculares e permitir a recuperação celular e a regeneração tecidual em dentes permanentes jovens com comprometimento endodôntico. Dois importantes flavonóis detectados no vinho tinto, morina (MO) e miricetina (MY), são atualmente estudados por suas amplas propriedades biológicas, incluindo atividade antimicrobiana. No entanto, o desenvolvimento de sistemas de liberação controlada pode ser útil para a liberação desses flavonóis para fins de terapia endodôntica. Este estudo avaliou a citocompatibilidade e os efeitos antimicrobianos/antibiofilme de MO e MY, isolados ou incorporados em hidrogéis termorreversíveis de quitosana-poloxamer-ß-glicerofosfato de sódio (CPG), além dos efeitos de MO e MY, isolados e combinados sobre a viabilidade, atividade de ALP e produção de nódulos de mineralização em células MDPC-23. A atividade antimicrobiana dos compostos foi avaliada em Streptococcus mutans, Enterococcus faecalis, Actinomyces israelii e Fusobacterium nucleatum em condições planctônicas, em biofilmes dual-espécies e multiespécies e analisadas por contagem bacteriana e microscopia de varredura. Os hidrogéis CPG foram caracterizados por reometria de fluxo e oscilatória, temperatura de gelificação, perfil de textura e análise de bioadesão em espécimes de dentina. MO, MY e controles (hidróxido de cálcio ­ CH e clorexidina ­ CHX) foram incorporados em hidrogéis de CPG e o efeito do antibiofilme sobre biofilmes multiespécies formados em amostras de dentina radicular foi avaliado por microscopia confocal. O efeito de toxicidade dos compostos isolados ou incorporados em hidrogéis de CGP foi determinado em cultura de fibroblastos por ensaios de resazurina. Os dados foram analisados estatisticamente pelos testes ANOVA e Tukey considerando p < 0,05. A combinação de MO e MY foi sinérgica ou aditiva contra bactérias endodônticas testadas a partir de concentrações de 0,03 mg/mL MO + 0,06 mg/mL MY e não foram tóxicas para fibroblastos até 0,125mg/mL. MO + MY apresentou melhor efeito sobre biofilmes dual-espécies e multiespécies considerando suas menores concentrações quando comparados com os flavonóis isolados. Os hidrogéis CPG foram caracterizados como termorreversíveis e com propriedades mecânicas e bioadesivas adequadas. Hidrogéis de CPG carregados com MO+MY, CH e CHX apresentaram efeitos inibitórios semelhantes quando aplicados em biofilmes multiespécies formados no interior dos túbulos dentinários radiculares por 48h e seus extratos apresentaram citotoxicidade acima de 50% de diluição. As células semelhantes a odontoblastos (MDPC-23) foram expostas a diferentes concentrações de MO, MY, isoladamente ou em combinação e CH como controle positivo por 24h e 48h, e troca contínua de meio osteogênico por 8 dias e 14 dias. As combinações de MO+MY ou CH também foram incorporadas em hidrogéis de quitosana-poloxamer-ß-glicerofosfato e seus extratos em meio de cultura celular foram coletados após 48h e 7 dias. Viabilidade celular, atividade de fosfatase alcalina (ALP) e ensaios de deposição de nódulos mineralizados (MN) foram realizados pelo método de resazurina, ensaios de monofosfato de timolftaleína e coloração com vermelho de alizarina, respectivamente. Todos os compostos não causaram citotoxicidade nas concentrações testadas em 24h e 8 dias e 0,5 mg/mL MO e MY isolados reduziram a viabilidade celular em 48h. A atividade de ALP e a deposição de MN foram aumentadas para a combinação MO+MY e CH em células MDPC-23. Extratos de hidrogel de 7 dias contendo ou não MO+MY não foram citotóxicos até diluição de 25% em 48h e em baixas concentrações estimularam a atividade de ALP e deposição de MN aos 8 e 14 dias de avaliação. Em conclusão, a combinação de morina e miricetina incorporada ou não em hidrogéis de CPG apresentou efeito antibiofilme sobre patógenos orais e baixa toxicidade sobre fibroblastos. Morina e miricetina em baixas concentrações, isoladas, em combinação ou em hidrogéis CPG não foram citotóxicas e foram eficazes na indução de marcadores de mineralização em células semelhantes a odontoblastos(AU)

A drug capable of disinfecting the root canals and allow cell recovery and tissue regeneration in permanent young teeth with endodontic problems has not been found yet. Two important flavonols detected in red wine, morin (MO) and myricetin (MY), are currently studied for their wide biological properties including antimicrobial activity. However, the development of controlled release systems could be useful for the delivery of these flavonols for endodontic therapies. This study evaluated the cytocompatibility and antimicrobial/antibiofilm effects of MO and MY, alone or incorporated in thermoreversible chitosanpoloxamer hydrogels containing sodium ß-glycerophosphate (CPG), in addition to the effects of isolated and combined morin and myricetin flavonols on viability, ALP activity and production of mineralization nodules in MDPC-23 cells. Antimicrobial activity of the compounds was evaluated on Streptococcus mutans, Enterococcus faecalis, Actinomyces israelii, and Fusobacterium nucleatum under planktonic conditions, on dual-species and multispecies biofilms and analyzed by bacterial counts and scanning microscopy. CPG hydrogels were characterized by flow and oscillatory rheometry, gelation temperature, texture profile and bioadhesion analysis in dentin specimens. MO, MY and controls (calcium hydroxide ­ CH and chlorhexidine ­ CHX) were incorporated in CPG hydrogels and antibiofilm effect on multispecies biofilms formed in radicular dentin samples were evaluated by confocal microscopy. Cytotoxicity of the compounds alone or incorporated in CGP hydrogels was determined on fibroblasts culture by resazurin assays. Data were statistically analyzed by ANOVA and Tukey considering p < 0.05. The combination of MO and MY had synergistic or additive against oral bacteria tested starting at concentrations of 0.03 mg/mL MO + 0.06 mg/mL MY and they were not toxic to fibroblasts up to 0.125mg/mL. MO + MY had better effect on dual-species and multispecies biofilms considering their lower concentrations when compared with the flavonols alone. CPG hydrogels were characterized as thermoreversible and with adequate mechanical and bioadhesive properties. CPG hydrogels loaded with MO+MY, CH and CHX have similar inhibitory effects when applied on multispecies biofilms formed inside root dentin tubules for 48h and their extracts were cytotoxicity above 50% dilution. Furthermore, the effects of morin, myricetin, alone or in combination or incorporated in chitosan-based hydrogels on cytotoxicity and expression of mineralization markers in odontoblast-like cells. The MDPC-23 cells were exposed to different concentrations of morin (MO), myricetin (MY), alone or in combination and calcium hydroxide (CH) as a positive control for 24h and 48h, and continuous osteogenic medium changing for 8 days and 14 days. The combinations of MO+MY or CH were also incorporated in chitosan-poloxamer-ß- glycerophosphate hydrogels and their extracts in cell culture media were collected after 48h and 7 days. Cell viability, alkaline phosphatase (ALP) activity and assays mineralized nodules (MN) deposition were performed using resazurin method, thymolphthalein monophosphate assays and alizarin red staining, respectively. Data were statistically analyzed considering p< 0.05. All compounds were non-toxic at the concentrations tested at 24h and 8 days and 0.5 mg/mL MO and MY alone reduced cell viability at 48h. ALP activity and deposition of MN were increased for MO+MY combination and CH in MDPC-23 cells. 7 days hydrogel extracts containing or not MO+MY were not cytotoxic up to 25% dilution at 48h and at low concentrations stimulated ALP activity and MN deposition at 8 and 14 days of evaluation. In conclusion, the combination of morin and myricetin incorporated or not in CPG hydrogels presented antibiofilm effect on oral pathogens and low cytotoxicity on fibroblasts. Morin myricetin at low concentrations, alone, in combination or in CPG hydrogels were not cytotoxic and were effective in inducing mineralization markers in odontoblast-like cells(AU)

Ethyl Acetate Fraction and Isolated Phenolics Derivatives from Mandevilla moricandiana Identified by UHPLC-DAD-ESI-MSn with Pharmacological Potential for the Improvement of Obesity-Induced Endothelial Dysfunction.

Endothelial dysfunction in obesity plays a key role in the development of cardiovascular diseases, and it is characterized by increased vascular tonus and oxidative stress. Thus, this study aimed to investigate the vasodilatory and antioxidant activities of Mandevilla moricandiana ethyl acetate fraction and subfractions. Vascular effects were investigated on aorta isolated from control and monosodium glutamate (MSG) induced-obese Wistar rats, and antioxidant activity was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) methods. The ethyl acetate fraction (MMEAF) induced a concentration-dependent vasodilation on aortic rings through the NO pathway, with the involvement of histamine H1 and estrogen ERα receptors and showed potent antioxidant activity. In aorta of MSG obese rats, maximal relaxation to acetylcholine was increased in the presence of MMEAF (3 µg/mL), indicating that MMEAF ameliorated obesity-induced endothelial dysfunction. Quercetin and kaempferol aglycones and their correspondent glycosides, as well as caffeoylquinic acid derivatives, A-type procyanidin trimer, ursolic and oleanolic triterpenoid acids were identified in subfractions from MMEAF and seem to be the metabolites responsible for the vascular and antioxidant activities of this fraction.

Extraction optimization of bioactive compounds from ora-pro-nobis (Pereskia aculeata Miller) leaves and their in vitro antioxidant and antihemolytic activities.

Ora-pro-nobis (Pereskia aculeata Miller) is a non-conventional food plant common in Brazil. The objective of this study was to optimize the extraction of bioactive phenolic compounds from ora-pro-nobis leaves by employing solvent mixtures. Ten extracts were obtained with water, ethanol, acetone, and their binary and ternary mixtures, evaluating the chemical composition, antioxidant activity and bioactivities in vitro. The response surface methodology was applied to model the results and calculate the optimal solvent composition, which is 60% water, 40% ethanol and 0% acetone. The optimized extract is rich in phenolic compounds (64 mg GAE/g) and proteins (823 mg/g) and presents antioxidant activity (in intracellular media as well) and inhibits lipid peroxidation (32%) along with hypotonic hemolysis (H50 = 0.339%), it does not present toxicity in vitro against cancer and normal cells. This is the first report of chicoric, caffeoyl-hexaric and coumaroyl-hexaric acids and some glycosylate derivatives of flavonols in ora-pro-nobis leaves.

Phenolics profiling by HPLC-DAD-ESI-MSn aided by principal component analysis to classify Rabbiteye and Highbush blueberries.

Although blueberries are widely studied, little information exists on their composition and content of flavonol glycosides. Most studies identify only a few flavonols in blueberries due to separation and identification issues. In the present study, we identified 44 flavonols and chlorogenic acid in 30 samples of Highbush and Rabbiteye blueberry, using HPLC-DAD-ESI-MSn. Highbush group fruits presented mainly quercetin-3-galactoside in their composition, while Rabbiteye group fruits exhibited higher levels of quercetin-3-rhamnoside and quercetin-3-glucuronide. Among the identified flavonols, 8 acylates (acetyl and hydroxymethylglutaroyl) were found, of which quercetin-3-O-[4″-(3-hydroxy-3-methylglutaroyl)]-α-rhamnoside was found for the first time in blueberries. This compound is exclusive to the cultivars Florida and Powderblue, where it is present in high quantities. Glucuronides of syringetin and laricitrin, and rhamnosyl-galactosides of myricetin, quercetin and isorhamnetin were also found for the first time in blueberries. The Principal Component Analysis showed that blueberry groups can be distinguished based on their phenolic compound profile.

Hypotensive activity of Campomanesia xanthocarpa leaf extract: beyond angiotensin II type 1 receptor blockage.

The ability of Campomanesia xanthocarpa leaf extract (CXLE) to alter blood pressure and heart rate was evaluated in anesthetized rats. The CXLE-induced hypotension was evaluated before and after losartan, methylatropine, L-N(ω)-nitro-L-arginine methyl ester (L-NAME), hexamethonium, indomethacin, glibenclamide, or nifedipine administration. The constituents of CXLE were identified by LC-DAD-MS. CXLE decreased blood pressure in a dose-dependent manner; only the highest dose decreased heart rate. The hypotension induced by CXLE was sensitive only to losartan, nifedipine, and glibenclamide. L-NAME decreased the time to recover 50% of the hypotensive effect of CXLE without altering its magnitude. Flavan-3-ols, proanthocyanidins (dimers and trimers), and glycosylated flavonols were identified from CXLE. The chemical constituents of CXLE seem to induce not only angiotensin II type 1 receptor blockage, but also ATP-sensitive potassium channels activation and L-type voltage-dependent Ca2+ channels inactivation. Nitric oxide is involved in the maintenance of the hypotensive effect of CXLE.

In vitro screening for acetylcholinesterase and butyrylcholinesterase inhibition and antimicrobial activity of chia seeds (Salvia hispanica)

BACKGROUND: Chia seeds are gaining increasing interest among food producers and consumers because of their prohealth properties. RESULTS: The aim of this work was to evaluate the potential of chia seeds to act as acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitors. The highest inhibitory activity against AChE and BChE was observed for colored seed ethanol extracts. A positive correlation was found between the presence of quercetin and isoquercetin as well as protocatechuic, hydroxybenzoic, and coumaric acids and the activity of extracts as AChE and BChE inhibitors. It has also been shown that grain fragmentation affects the increase in the activity of seeds against cholinesterases (ChE). Furthermore, seeds have been shown to be a source of substances that inhibit microbial growth. CONCLUSIONS: It was found that the chia seed extracts are rich in polyphenols and inhibit the activity of ChEs; therefore, their use can be considered in further research in the field of treatment and prevention of neurodegenerative diseases.

Effects of Flavonoids and Its Derivatives on Immune Cell Responses.

BACKGROUND: Various pieces of evidence have shown that people who consume foods rich in polyphenolic and flavonoids compounds have a lower incidence of inflammatory, autoimmune diseases and cancer. OBJECTIVE: The study aimed to review the most potent compounds that affect the immune response and diseases associated with it. METHODS: Publications in PubMed and EmBase, from 1974-2018, and patents form Free patents online, Scifinder, Espacenet and Mendeley in which flavonoids, their semi-synthetic and synthetic derivatives are involved in immunosuppressive or immunostimulatory responses in vitro and in vivo. RESULTS: In vitro, flavonoids and their derivatives inhibit various transcriptional factors, which modulate differentiation, proliferation, activation of immune cells and enhance regulatory T cell generation. Some flavonoids exert anti-inflammatory effects through: Blockade of NF-κB, and NLRP3 inflammasome, inhibition of pro-inflammatory cytokine production, IL-1ß, IL-2, IL-6, TNF-α, IL-17A, down regulation of chemokines, and reduction of reactive oxygen and nitrogen species. Nevertheless, several reports have shown that some flavonoids enhance immune response by enhancing: oxygen and nitrogen radicals, antibody production, cytotoxic activity against tumours by increasing activating receptors and down regulating inhibitory receptors. In consequence, flavonoids may be potentially useful for treatment of infectious diseases and cancer. CONCLUSION: The most potent flavonoids in inflammation that modify immune responses are apigenin, quercetin and Epigallocatechin-3-Gallate (EGCG) although, other compounds are still under study and cannot be excluded. The most relevant patents concerning the use of flavones and other polyphenols were revised. A promising future of these compounds in different therapies is discussed.