RESUMO
Developmental biology has greatly profited from genetic and reverse genetic approaches to indirectly studying protein function. More recently, nanobodies and other protein binders derived from different synthetic scaffolds have been used to directly dissect protein function. Protein binders have been fused to functional domains, such as to lead to protein degradation, relocalization, visualization, or posttranslational modification of the target protein upon binding. The use of such functionalized protein binders has allowed the study of the proteome during development in an unprecedented manner. In the coming years, the advent of the computational design of protein binders, together with further advances in scaffold engineering and synthetic biology, will fuel the development of novel protein binder-based technologies. Studying the proteome with increased precision will contribute to a better understanding of the immense molecular complexities hidden each step along the way to generate form and function during development.
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Hedgehog (Hh) signal molecules play a fundamental role in development, adult stem cell maintenance and cancer. Hh can signal at a distance, and we have proposed that its graded distribution across Drosophila epithelia is mediated by filopodia-like structures called cytonemes. Hh reception by Patched (Ptc) happens at discrete sites along presenting and receiving cytonemes, reminiscent of synaptic processes. Here, we show that a vesicle fusion mechanism mediated by SNARE proteins is required for Ptc placement at contact sites. Transport of Ptc to these sites requires multivesicular bodies (MVBs) formation via ESCRT machinery, in a manner different to that regulating Ptc/Hh lysosomal degradation after reception. These MVBs include extracellular vesicle (EV) markers and, accordingly, Ptc is detected in the purified exosomal fraction from cultured cells. Blockage of Ptc trafficking and fusion to basolateral membranes result in low levels of Ptc presentation for reception, causing an extended and flattened Hh gradient.
Assuntos
Proteínas de Drosophila/metabolismo , Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Proteínas Hedgehog/metabolismo , Discos Imaginais/metabolismo , Receptores de Superfície Celular/metabolismo , Proteínas SNARE/metabolismo , Asas de Animais , Animais , Proteínas de Drosophila/genética , Drosophila melanogaster , Complexos Endossomais de Distribuição Requeridos para Transporte/genética , Proteínas Hedgehog/genética , Transporte Proteico , Receptores de Superfície Celular/genética , Proteínas SNARE/genéticaRESUMO
Cellular development and function rely on highly dynamic molecular interactions among proteins distributed in all cell compartments. Analysis of these interactions has been one of the main topics in cellular and developmental research, and has been mostly achieved by the manipulation of proteins of interest (POIs) at the genetic level. Although genetic strategies have significantly contributed to our current understanding, targeting specific interactions of POIs in a time- and space-controlled manner or analysing the role of POIs in dynamic cellular processes, such as cell migration or cell division, would benefit from more-direct approaches. The recent development of specific protein binders, which can be expressed and function intracellularly, along with advancement in synthetic biology, have contributed to the creation of a new toolbox for direct protein manipulations. Here, we have selected a number of short-tag epitopes for which protein binders from different scaffolds have been generated and showed that single copies of these tags allowed efficient POI binding and manipulation in living cells. Using Drosophila, we also find that single short tags can be used for POI manipulation in vivo.
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Drosophila melanogaster/genética , Epitopos/genética , Peptídeos/genética , Proteínas/genética , Animais , Linhagem Celular , Células Cultivadas , Peptídeos/química , Ligação Proteica/genética , Proteínas/química , Biologia SintéticaRESUMO
OBJECTIVE: To develop N-(levodopa) chitosan derivatives through click chemistry to study their effect in brain cells.Significance: This study presents a proof-of-concept that macromolecules such as N-(Levodopa) chitosan derivatives traverse brain cell membranes and induce biomedical functionalities. METHODS: Through click chemistry, we developed N-(levodopa) chitosan derivatives. They were physically and chemically characterized by FT-IR, 1H-NMR, TGA and Dynamic Light Scattering analyses. Solution and nanoparticles of N-(levodopa) chitosan derivatives were tested in primary cell cultures from the postnatal rat olfactory bulb, substantia nigra and corpus callosum. Ca2+ imaging and UPLC experiments were used to investigate if the biomaterial modulated the brain cell physiology. RESULTS: N-(levodopa) chitosan derivatives induced intracellular Ca2+ responses in primary cell cultures of the rat brain. UPLC experiments indicated that levodopa attached to chitosan was converted into dopamine by brain cells. CONCLUSION: The present study shows that N-(levodopa) chitosan may be useful to develop new treatment strategies, which could serve as molecular reservoirs of biomedical drugs to treat degenerative disorders of the nervous system.
Assuntos
Quitosana , Levodopa , Ratos , Animais , Levodopa/farmacologia , Quitosana/química , Química Click/métodos , Espectroscopia de Infravermelho com Transformada de Fourier , EncéfaloRESUMO
Obesity is a serious health problem worldwide, since it is associated with multiple metabolic disorders and complications such as cardiovascular disease, type 2 diabetes, fatty liver disease and overall metabolic dysfunction. Dysregulation of the hunger-satiety pathway, which includes alterations of central and peripheral signaling, explains some forms of obesity by favoring hyperphagia and weight gain. The present work comprehensively summarizes the mechanisms by which naringenin (NAR), a predominant flavanone in citrus fruits, could modulate the main pathways associated with the development of obesity and some of its comorbidities, such as oxidative stress (OS), inflammation, insulin resistance (IR) and dyslipidemia, as well as the role of NAR in modulating the secretion of enterohormones of the satiety pathway and its possible antiobesogenic effect. The results of multiple in vitro and in vivo studies have shown that NAR has various potentially modulatory biological effects against obesity by countering IR, inflammation, OS, macrophage infiltration, dyslipidemia, hepatic steatosis, and adipose deposition. Likewise, NAR is capable of modulating peptides or peripheral hormones directly associated with the hunger-satiety pathway, such as ghrelin, cholecystokinin, insulin, adiponectin and leptin. The evidence supports the use of NAR as a promising alternative to prevent overweight and obesity.
Assuntos
Diabetes Mellitus Tipo 2 , Flavanonas , Resistência à Insulina , Hepatopatia Gordurosa não Alcoólica , Humanos , Diabetes Mellitus Tipo 2/complicações , Obesidade/metabolismo , Flavanonas/farmacologia , Inflamação/tratamento farmacológico , Inflamação/complicações , Hepatopatia Gordurosa não Alcoólica/complicaçõesRESUMO
BACKGROUND: Probiotics and antispasmodics have been tested extensively in the management of symptoms of irritable bowel syndrome (IBS), but they have rarely been evaluated in combination. The objective of this pilot study was to assess the efficacy of treatment with the probiotic formulation i3.1 (Lactobacillus plantarum CECT7484 and CECT7485 and Pediococcus acidilactici CECT7483), with or without the addition of the antispasmodic alverine/simethicone, in improving IBS-related quality of life (QoL) and reducing abdominal pain and diarrhea in patients with IBS. METHODS: This was a randomized, placebo-controlled clinical trial with 3 parallel arms (probiotic, probiotic plus antispasmodic, and placebo). Patients with IBS (N = 55) were recruited at the Gastroenterology Department of the Juárez Hospital (México City). QoL was assessed with the IBS-QoL questionnaire, abdominal pain with a visual analog scale, and stool consistency with the Bristol scale. RESULTS: The IBS-QoL rate of response (ITT analysis) was 50.0% for patients in the group with probiotic alone, 68.4% in the group with probiotic plus antispasmodic, and 16.7% in the group with placebo after 6 weeks of treatment (p = 0.005). Response to abdominal pain was reported by 38.9% of patients treated with probiotic, 57.9% with probiotic plus antispasmodic, and 16.7% with placebo (p = 0.035). Regarding stool consistency, a response to treatment was reported by 44.4% of patients treated with probiotic, 57.9% with probiotic plus antispasmodic, and 16.7% with placebo (p = 0.032). CONCLUSION: The results are consistent with previous studies on the use of the i3.1 probiotic formulation for the management of symptoms in IBS patients, and the addition of an antispasmodic improves its observed effects.
Assuntos
Síndrome do Intestino Irritável/tratamento farmacológico , Parassimpatolíticos/uso terapêutico , Probióticos/uso terapêutico , Qualidade de Vida , Dor Abdominal/tratamento farmacológico , Adulto , Método Duplo-Cego , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Probióticos/administração & dosagem , Inquéritos e Questionários , Resultado do TratamentoRESUMO
The role of non-energy-yielding nutrients on health has been meticulously studied, and the evidence shows that a compound can exert significant effects on health even if not strictly required by the organism. Phenolic compounds are among the most widely studied molecules that fit this description; they are found in plants as secondary metabolites and are not required by humans for growth or development, but they can influence a wide array of processes that modulate health across multiple organs and systems. The lower gastrointestinal tract is a prime site of action of phenolic compounds, namely, by their effects on gut microbiota and colonic health. As with humans, phenolic compounds are not required by most bacteria but can be substrates of others; in fact, some phenolic compounds exert antibacterial actions. A diet rich in phenolic compounds can lead to qualitative and quantitative effects on gut microbiota, thereby inducing indirect health effects in mammals through the action of these microorganisms. Moreover, phenolic compounds may be fermented by the gut microbiota, thereby modulating the compounds bioactivity. In the colon, phenolic compounds promote anti-inflammatory, anti-oxidant and antiproliferative actions. The aim of the present review is to highlight the role of phenolic compounds on maintaining or restoring a healthy microbiota and overall colonic health. Mechanisms of action that substantiate the reported evidence will also be discussed.
Assuntos
Bactérias/efeitos dos fármacos , Colo/microbiologia , Colo/fisiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Fenóis/farmacologia , Bactérias/classificação , Dieta , HumanosRESUMO
The meninges shield the nervous system from diverse, rather harmful stimuli and pathogens from the periphery. This tissue is composed of brain endothelial cells (BECs) that express diverse ion channels and chemical-transmitter receptors also expressed by neurons and glial cells to communicate with each other. However, information about the effects of ATP and angiotensin II on BECs is scarce, despite their essential roles in blood physiology. This work investigated in vitro if BECs from the meninges from rat forebrain respond to ATP, angiotensin II and high extracellular potassium, with intracellular calcium mobilizations and its second messenger-associated pathways. We found that in primary BEC cultures, both ATP and angiotensin II produced intracellular calcium responses linked to the activation of inositol trisphosphate receptors and ryanodine receptors, which led to calcium release from intracellular stores. We also used RT-PCR to explore what potassium channel subunits are expressed by primary BEC cultures and freshly isolated meningeal tissue, and which might be linked to the observed effects. We found that BECs mainly expressed the inward rectifier potassium channel subunits Kir1.1, Kir3.3, Kir 4.1 and Kir6.2. This study contributes to the understanding of the functions elicited by ATP and angiotensin II in BECs from rat meninges. SIGNIFICANCE OF THE STUDY: Brain endothelial cells (BECs) express diverse ion channels and membrane receptors, which they might use to communicate with neurons and glia. This work investigated in vitro, if BECs from the rat forebrain respond to angiotensin II and ATP with intracellular calcium mobilizations. We found that these cells did respond to said substances with intracellular calcium mobilizations linked to inositol trisphosphate and ryanodine receptor activation, which led to calcium release from intracellular stores. These findings are important because they might uncover routes of active communication between brain cells and endothelial cells.
Assuntos
Trifosfato de Adenosina/farmacologia , Angiotensina II/farmacologia , Cálcio/metabolismo , Células Endoteliais/efeitos dos fármacos , Potássio/farmacologia , Prosencéfalo/metabolismo , Animais , Células Cultivadas , Células Endoteliais/metabolismo , Feminino , Masculino , Canais de Potássio/genética , Canais de Potássio/metabolismo , Prosencéfalo/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
Neolentinus lepideus is a fungus consumed by rural communities in Central America and Asia due to its rich flavor; however, little information on its chemical composition is available. With this in mind, the objective of this work was to determine the content of vitaminâ E and C, ergosterol, and phenolic compounds of this fungus, as well as its antioxidant capacity. The quantified bioactive compounds were two isoforms of vitaminâ E, highlighting α-tocopherol (3370.35â mg/100â g dry weight, DW) and ergosterol (11.70â mg/100â g DW). The total phenolic content was 164.80â mg gallic acid equivalents/100â g, and nine phenolic compounds were identified (protocatechuic, p-hydroxybenzoic, caffeic, vanillic, ferulic, salicylic, p-anisic, trans-cinnamic acids, and scopoletin). The highest antioxidant capacity was detected in the lipophilic extract with TEAC (27688â µmoles Trolox equivalents/100â g). These results suggest that lipophilic compounds are among the main bioactive compounds in N. lepideus, and they might exhibit the highest radical scavenging properties in non-polar extracts.
Assuntos
Antioxidantes/farmacologia , Basidiomycota/química , Cromanos/antagonistas & inibidores , Antioxidantes/química , Antioxidantes/isolamento & purificaçãoRESUMO
Lactic acid fermentation increases the bioactive properties of shrimp waste. Astaxanthin is the principal carotenoid present in shrimp waste, which can be found esterified in the liquid fraction (liquor) after its lactic acid fermentation. Supercritical CO2 technology has been proposed as a green alternative to obtain astaxanthin from fermented shrimp waste. This study aimed to optimize astaxanthin extraction by supercritical CO2 technology from fermented liquor of shrimp waste and study bioaccessibility using simulated gastrointestinal digestion (GD) of the optimized extract. A Box-Behnken design with three variables (pressure, temperature, and flow rate) was used to optimize the supercritical CO2 extraction. The optimized CO2 extract was obtained at 300 bar, 60 °C, and 6 mL/min, and the estimated characteristics showed a predictive extraction yield of 11.17%, antioxidant capacity of 1.965 mmol of Trolox equivalent (TE)/g, and astaxanthin concentration of 0.6353 µg/g. The experiment with optimal conditions performed to validate the predicted values showed an extraction yield of 12.62%, an antioxidant capacity of 1.784 mmol TE/g, and an astaxanthin concentration of 0.52 µg/g. The astaxanthin concentration decreased, and the antioxidant capacity of the optimized extract increased during gastrointestinal digestion. In conclusion, our optimized supercritical CO2 process is suitable for obtaining astaxanthin from shrimp by-products after lactic acid fermentation.
Assuntos
Antioxidantes , Penaeidae/química , Animais , Antioxidantes/análise , Antioxidantes/isolamento & purificação , Dióxido de Carbono/química , Fermentação , Resíduos , Xantofilas/análise , Xantofilas/isolamento & purificaçãoRESUMO
BACKGROUND: Oxidative stress has been implicated in the pathogenesis and progression of diabetes mellitus. Both can damage the brain. Mango and its by-products are sources of bioactive compounds with antioxidant properties. We hypothesized that mango cv. 'Ataulfo' peel and pulp mitigate oxidative stress in the brain of streptozotocin-induced diabetic rats. RESULTS: Twenty-four male Wistar rats were divided into four groups: control, untreated diabetic (UD), diabetic treated with a mango-supplemented diet (MTD), and diabetic pretreated with a mango-supplemented diet (MPD). The rats were fed the different diets for 4 weeks after diabetes induction (MTD), or 2 weeks before and 4 weeks after induction (MPD). After the intervention, serum and brain (cerebellum and cortex) were collected to evaluate gene expression, enzyme activity, and redox biomarkers. Superoxide dismutase 2 (SOD2) expression increased in the cortex of the MTD group, whereas glutathione-S-transferase p1 (GSTp1) expression was higher in the cortex of the MTD group, and cortex and cerebellum of the MPD group. SOD1 activity was higher in the cerebellum and cortex of all diabetic groups, whereas GST activity increased in the cerebellum and cortex of the MPD group. Lipid peroxidation increased in the cerebellum and cortex of the UD group; however, a mango-supplemented diet prevented this increase in both regions, while also mitigating polyphagia and weight loss, and maintaining stable glycemia in diabetic rats. CONCLUSION: We propose that mango exerts potent neuroprotective properties against diabetes-induced oxidative stress. It can be an alternative to prevent and treat biochemical alterations caused by diabetes. © 2020 Society of Chemical Industry.
Assuntos
Diabetes Mellitus/tratamento farmacológico , Mangifera/química , Fármacos Neuroprotetores/administração & dosagem , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/administração & dosagem , Animais , Glicemia/metabolismo , Cerebelo/efeitos dos fármacos , Cerebelo/metabolismo , Diabetes Mellitus/genética , Diabetes Mellitus/metabolismo , Frutas/química , Glutationa/metabolismo , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , Estreptozocina , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismoRESUMO
OBJECTIVE: To report the species identified in candidemia isolated in hospitals in Paraguay, their distribution by age groups, and their susceptibility to fluconazole, voriconazole, and amphotericin B. METHODS: Retrospective study of candidemia reported to the Central Health Public Laboratory by seven hospitals between 2010 and 2018. Identification and susceptibility testing were performed using the automated Vitek 2® system. RESULTS: 520 cases of candidiasis were reported. The prevalent species were Candida albicans (34.4%), C. parapsilosis (30.4%), C. tropical (25.4%), C. glabrata (4.8%), and C. krusei (2.1%). Less frequent species were: C. lusitaniae (1.2%), C. guilliermondii (1%), C. famata (0.3%), C. rugous (0.2%), and C. kefyr (0.2%). Candidiasis was most prevalent in the elderly (48.5%) and adults (29.9%). C. parapsilosis was most frequent in newborns and infants, and C. tropical in pediatric patients; 2.8% and 0.6% of C. albicans and 4.4% and 2.5% of C. parapsilosis were resistant, respectively, to fluconazole and voriconazole; 8% of C. glabrata were resistant to fluconazole and two isolations showed a minimum inhibitory concentration for voriconazole greater than 0.25 mg/L. CONCLUSIONS: This is the first report on candidemias in hospitals of Paraguay. Together with other studies, it will improve knowledge about this infection in the country.
OBJETIVO: Relatar as espécies identificadas em candidemias isoladas em hospitais do país, apresentar a sua distribuição por faixa etária e determinar a sua sensibilidade ao fluconazol, voriconazol e anfotericina B. MÉTODOS: Estudo retrospectivo de candidemias notificadas por sete hospitais ao Laboratório Central de Saúde Pública no período de 2010 a 2018. A identificação e o antifungigrama foram realizados pelo sistema automatizado Vitek®2. RESULTADOS: Foram notificados 520 casos de candidemia. As espécies predominantes foram Candida albicans (34,4%), C. parapsilosis (30,4%), C. tropicalis (25,4%), C. glabrata (4,8%) e C. krusei (2,1%). Outras espécies menos frequentes foram: C. lusitaniae (1,2%), C. guilliermondii (1%), C. famata (0,3%), C. rugosa (0,2%) e C. kefyr (0,2%). As candidemias em pacientes idosos (48,5%) e adultos (29,9%) foram as mais prevalentes. C. parapsilosis foi a espécie mais frequente em recém-nascidos e lactentes, e C. tropicalis predominou em pacientes pediátricos. Ao todo, 2,8% e 0,6% dos casos de C. albicans e 4,4% e 2,5% dos de C. parapsilosis foram resistentes a fluconazol e voriconazol, respectivamente. Além disso, 8% dos casos de C. glabrata foram resistentes ao fluconazol, e duas amostras isoladas apresentaram uma concentração inibitória mínima para voriconazol acima de 0,25 mg/L. CONCLUSÕES: Este é o primeiro estudo sobre candidemias em hospitais do Paraguai, o que, juntamente com outros estudos, permitirá conhecer melhor esta infecção no país.
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Onion waste disposal issue could be solved by using onion skin as food ingredient. Therefore, the aim of present study is the estimation of flavonoid concentration, total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activities of onion skin of fifteen Indian cultivars. Flavonoid quantification was achieved by high performance liquid chromatography, which showed highest concentration of quercetin, quercetin 3-ß-D-glucoside, luteolin and kaempferol in cv. 'NHRDF Red' (11,885.025 mg/kg), 'Hissar-2' (1432.875 mg/kg), 'Pusa Riddhi' (1669.925 mg/kg) and 'Bhima Shakti' (709.975 mg/kg), respectively in dry weight. Highest TPC and TFC were found in cv. 'NHRDF Red' while lowest were measured in cv. 'Bhima Shubhra'. DPPH assay (%), ABTS assay (%) and FRAP assay (µmol gallic acid/g) were showed maximum antioxidant capacity for cv. 'NHRDF Red' whereas least obtained for cv. 'Bhima Shubhra'. Skin of cv. 'Hissar-2' and 'NHRDF Red' are the best source of flavonoids and natural antioxidants.
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The present study evaluated the contribution of mango fiber (MF) and mango phenolic compounds (MP) to the hepatoprotective effect of freeze-dried mango pulp (FDM) cultivar (cv.) "Ataulfo" diets in high cholesterol/sodium cholate (HCC)-fed rats. Male Wistar rats were fed with a HCC diet for 12 weeks, either untreated, or supplemented with MF, MP, FDM, or a control diet (no HCC; n = 6/group). All mango treatments significantly decreased hepatic cholesterol deposition and altered its fatty acid profile, whereas MF and MP mitigated adipose tissue hypertrophy. MF caused a lower level of proinflammatory cytokines (IL-1α/ß, IFN-γ, TNF-α) whereas FDM increased the anti-inflammatory ones (IL-4, 6, 10). Mango treatments increased catalase (CAT) activity and its mRNA expression; superoxide dismutase (SOD) activity was normalized by MF and FDM, but its activity was unrelated to its hepatic mRNA expression. Changes in CAT and SOD mRNA expression were unrelated to altered Nrf2 mRNA expression. Higher hepatic PPARα and LXRα mRNA levels were found in MP and MF. We concluded that MF and MP are highly bioactive, according to the documented hepatoprotection in HCC-fed rats; their mechanism of action appears to be related to modulating cholesterol and fatty acid metabolism as well as to stimulating the endogenous antioxidant system.
Assuntos
Citoproteção/efeitos dos fármacos , Fibras na Dieta/farmacologia , Fígado/efeitos dos fármacos , Mangifera/química , Hepatopatia Gordurosa não Alcoólica/prevenção & controle , Fenóis/farmacologia , Animais , Antioxidantes/farmacologia , Colesterol na Dieta/administração & dosagem , Colesterol na Dieta/efeitos adversos , Dieta/efeitos adversos , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Hipercolesterolemia/induzido quimicamente , Hipercolesterolemia/prevenção & controle , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Hepatopatia Gordurosa não Alcoólica/patologia , PPAR alfa/metabolismo , Fenóis/isolamento & purificação , Ratos , Ratos Wistar , Colato de Sódio/administração & dosagem , Colato de Sódio/efeitos adversosRESUMO
BACKGROUND: Serum lipoproteins are in dynamic equilibrium, partially controlled by the apolipoprotein A1 to apolipoprotein B ratio (APOA1/APOB). Freeze-dried mango pulp (FDM) is a rich source of phenolic compounds (MP) and dietary fiber (MF), although their effects on lipoprotein metabolism have not yet been studied. RESULTS: Thirty male Wistar rats were fed with four different isocaloric diets (3.4 kcal g-1 ) for 12 weeks: control diet, high cholesterol (8 g kg-1 ) + sodium cholate (2 g kg-1 ) diet either alone or supplemented with MF (60 g kg-1 ), MP (1 g kg-1 ) or FDM (50 g kg-1 ). MP and FDM reduced food intake, whereas MF and MP tended to increase serum APOA1/APOB ratio, independently of their hepatic gene expression. This suggests that lipoprotein metabolism was favorably altered by mango bioactives, MP also mitigated the non-alcoholic steatohepatitis that resulted from the intake of this diet. CONCLUSION: We propose that phenolics are the most bioactive components of mango pulp, acting as anti-atherogenic and hepatoprotective agents, with a mechanism of action tentatively based on changes to the main protein components of lipoproteins. © 2018 Society of Chemical Industry.
Assuntos
Apolipoproteína A-I/sangue , Apolipoproteínas B/sangue , Colesterol/metabolismo , Mangifera/metabolismo , Hepatopatia Gordurosa não Alcoólica/dietoterapia , Fenol/metabolismo , Extratos Vegetais/administração & dosagem , Extratos Vegetais/metabolismo , Colato de Sódio/metabolismo , Animais , Humanos , Fígado/metabolismo , Masculino , Mangifera/química , Hepatopatia Gordurosa não Alcoólica/metabolismo , Fenol/análise , Extratos Vegetais/análise , Ratos , Ratos WistarRESUMO
Fruit ripening induces changes that strongly affect their matrices, and consequently, the bioaccessibility/bioavailability of its phenolic compounds. Flesh from 'slightly' (SR), 'moderately' (MR) and 'fully' (FR) ripe 'Ataulfo' mangoes were physicochemically characterized, and digested in vitro to evaluate how ripening impacts the bioaccessibility/bioavailability of its phenolic compounds. Ripening increased the flesh's pH and total soluble solids, while decreasing citric acid, malic acid and titratable acidity. MR and FR mango phenolics had higher bioaccessibility/bioavailability, which was related to a decreased starch and dietary fiber (soluble and insoluble) content. These results suggest that phenolics are strongly bound to the fruit's matrix of SR mango, but ripening liberates them as the major polysaccharides are hydrolyzed, thus breaking covalent bonds and disrupting carbohydrate-phenolic complexes. There was also a higher release percentage in the gastric digestion phase, as compared to the intestinal. Our data showed that the bioaccessibility/bioavailability of mango phenolics depends on fruit ripening and on digestion phase.
RESUMO
Mango (Mangifera indica cv. Ataulfo) peel contains bound phenolics that may be released by alkaline or acid hydrolysis and may be converted into less complex molecules. Free phenolics from mango cv. Ataulfo peel were obtained using a methanolic extraction, and their cellular antioxidant activity (CAA) and permeability were compared to those obtained for bound phenolics released by alkaline or acid hydrolysis. Gallic acid was found as a simple phenolic acid after alkaline hydrolysis along with mangiferin isomers and quercetin as aglycone and glycosides. Only gallic acid, ethyl gallate, mangiferin, and quercetin were identified in the acid fraction. The acid and alkaline fractions showed the highest CAA (60.5% and 51.5%) when tested at 125 µg/mL. The value of the apparent permeability coefficient (Papp) across the Caco-2/HT-29 monolayer of gallic acid from the alkaline fraction was higher (2.61 × 10-6 cm/s) than in the other fractions and similar to that obtained when tested pure (2.48 × 10-6 cm/s). In conclusion, mango peels contain bound phenolic compounds that, after their release, have permeability similar to pure compounds and exert an important CAA. This finding can be applied in the development of nutraceuticals using this important by-product from the mango processing industry.
Assuntos
Antioxidantes/farmacologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Mangifera/química , Fenóis/farmacologia , Extratos Vegetais/farmacologia , Antioxidantes/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Humanos , Permeabilidade , Fenóis/química , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/químicaRESUMO
Mango "Ataulfo" peel is a rich source of polyphenols (PP), with antioxidant and anti-cancer properties; however, it is unknown whether such antiproliferative activity is related to PP's antioxidant activity. The content (HPLC-DAD), antioxidant (DPPH, FRAP, ORAC), and antiproliferative activities (MTT) of free (FP) and chemically-released PP from mango 'Ataulfo' peel after alkaline (AKP) and acid (AP) hydrolysis, were evaluated. AKP fraction was higher (µg/g DW) in gallic acid (GA; 23,816 ± 284) than AP (5610 ± 8) of FR (not detected) fractions. AKP fraction and GA showed the highest antioxidant activity (DPPH/FRAP/ORAC) and GA's antioxidant activity follows a single electron transfer (SET) mechanism. AKP and GA also showed the best antiproliferative activity against human colon adenocarcinoma cells (LS180; IC50 (µg/mL) 138.2 ± 2.5 and 45.7 ± 5.2) and mouse connective cells (L929; 93.5 ± 7.7 and 65.3 ± 1.2); Cheminformatics confirmed the hydrophilic nature (LogP, 0.6) and a good absorption capacity (75%) for GA. Data suggests that GA's antiproliferative activity appears to be related to its antioxidant mechanism, although other mechanisms after its absorption could also be involved.
Assuntos
Antioxidantes/farmacologia , Ácido Gálico/análise , Mangifera/química , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Humanos , Extratos Vegetais/química , Polifenóis/farmacologiaRESUMO
The objective of this study was to evaluate the effect of combining catechin, protocatechuic, and vanillic acids against planktonic growing, adhesion, and biofilm eradication of uropathogenic Escherichia coli (UPEC), as well as antioxidant agents. The minimum inhibitory concentrations (MIC) of protocatechuic, vanillic acids and catechin against the growth of planktonic bacteria were 12.98, 11.80, and 13.78 mM, respectively. Mixing 1.62 mM protocatechuic acid + 0.74 mM vanillic acid + 0.05 mM catechin resulted in a synergistic effect acting as an MIC. Similarly, the minimum concentrations of phenolic compounds to prevent UPEC adhesion and biofilm formation (MBIC) were 11.03 and 7.13 mM of protocatechuic and vanillic acids, respectively, whereas no MBIC of catechin was found. However, combinations of 1.62 mM protocatechuic acid + 0.74 mM vanillic acid + 0.05 mM catechin showed a synergistic effect acting as MBIC. On the other hand, the minimum concentrations to eradicate biofilms (MBEC) were 25.95 and 23.78 mM, respectively. The combination of 3.20 mM protocatechuic acid, 2.97 mM vanillic acid, and 1.72 mM catechin eradicated pre-formed biofilms. The antioxidant capacity of the combination of phenolics was higher than the expected theoretical values, indicating synergism by the DPPHâ¢, ABTS, and FRAP assays. Effective concentrations of catechin, protocatechuic, and vanillic acids were reduced from 8 to 1378 times when combined. In contrast, the antibiotic nitrofurantoin was not effective in eradicating biofilms from silicone surfaces. In conclusion, the mixture of phenolic compounds was more effective in preventing cell adhesion and eradicating pre-formed biofilms of uropathogenic E. coli than single compounds and nitrofurantoin, and showed antioxidant synergy.
Assuntos
Antibacterianos/farmacologia , Catequina/farmacologia , Hidroxibenzoatos/farmacologia , Ácido Vanílico/farmacologia , Antibacterianos/química , Antioxidantes/química , Antioxidantes/farmacologia , Biofilmes/efeitos dos fármacos , Catequina/química , Humanos , Hidroxibenzoatos/química , Testes de Sensibilidade Microbiana , Plâncton/efeitos dos fármacos , Escherichia coli Uropatogênica/efeitos dos fármacos , Escherichia coli Uropatogênica/patogenicidade , Ácido Vanílico/químicaRESUMO
Agave leaves are considered a by-product of alcoholic beverage production (tequila, mezcal and bacanora) because they are discarded during the production process, despite accounting for approximately 50% of the total plant weight. These by-products constitute a potential source of Agave extracts rich in bioactive compounds, such as saponins, phenolic compounds and terpenes, and possess different biological effects, as demonstrated by in vitro and in vivo tests (e.g. antimicrobial, antifungal, antioxidant, anti-inflammatory, antihypertensive, immunomodulatory, antiparasitic and anticancer activity). Despite their positive results in biological assays, Agave extracts have not been widely evaluated in food systems and pharmaceutical areas, and these fields represent a potential route to improve the usage of Agave plants as food additives and agents for treating medical diseases. © 2017 Society of Chemical Industry.