GC/MS and 2D NMR-based approach to evaluate the chemical profile of hydroalcoholic extract from Agaricus blazei Murill and its anti-inflammatory effect on human neutrophils.

ETHNOPHARMACOLOGICAL RELEVANCE: Agaricus blazei Murill (AbM) is one of the main mushrooms used for medicinal purposes. The use of AbM in the preparation of teas is widespread mainly in Asian countries, while in Brazil it is used as a functional food to combat inflammatory diseases and cancer. AIM OF THE STUDY: The main focus of this study was the characterization of the chemical profile of the hydroalcoholic extract of Agaricus blazei Murill (AbE), as well as the evaluation of its cytotoxic and anti-inflammatory potential using human neutrophils. MATERIALS AND METHODS: The extract was prepared by dynamic maceration using a mixture of ethanol and water (70/30, v v-1) as solvent. The chemical profile characterization was carried out by 2D NMR and GC-MS techniques. The cytotoxicity of AbE was evaluated through studies of hemolytic potential, cell viability and membrane integrity. The anti-inflammatory activity was analyzed by a PMA-induced neutrophil degranulation assay. RESULTS: Chemical analysis of AbE revealed the presence of 28 metabolites in its composition, with mannitol as the major compound. AbE at 1-200 µg mL-1 and mannitol at 4-160 µg mL-1, showed low hemolytic and cytotoxic potential against human red blood cells and neutrophils. Furthermore, both were able to significantly reduce the release of myeloperoxidase. CONCLUSIONS: These results indicate that AbE is a promising natural product to be incorporated into pharmaceutical dosage forms intended for the adjuvant treatment of inflammatory diseases.

Effect of Methyl Jasmonate on the Biosynthesis of Volatile Compounds Associated with the Ripening of Grape Tomato Fruits.

The present work aims to evaluate the roles of methyl jasmonate (MeJA) in the formation of volatile organic compounds (VOC) from grape tomatoes during ripening. Fruits were treated with MeJA, ethylene, 1-MCP (1-methylcyclopropene), and MeJA+1-MCP, with analyses of the VOC and levels of the gene transcripts for the enzymes lipoxygenase (LOX), alcohol dehydrogenase (ADH), and hydroperoxide lyase (HPL). An intimate relationship between MeJA and ethylene in aroma formation was detected, mainly among the VOC from the carotenoid pathway. Expression of the fatty acid transcripts, LOXC, ADH, and HPL pathway genes, was reduced by 1-MCP, even when associated with MeJA. In ripe tomato, MeJA increased most of the volatile C6 compounds, except 1-hexanol. The MeJA+1-MCP treatment followed most of the increases in volatile C6 compounds that were increased by MeJA alone, which evidenced some ethylene-independent mechanism in the production of the volatile C6 compounds. In ripe tomato, MeJA and MeJA+1-MCP increased the levels of 6-methyl-5-hepten-2-one, which is derived from lycopene, evidencing an ethylene-independent biosynthetic process.

UPLC-QTOF-MS/MS-based metabolomic approach and gastroprotective effect of two chemotypes of Egletes viscosa (L.) less. against ethanol-induced gastric ulcer in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Egletes viscosa (L.) (macela) is a native wild herb that can be found in different states of northeastern Brazil. The infusions of its flower buds are traditionally used for the treatment of gastrointestinal disorders. E. viscosa possesses two chemotypes (named A and B), distinguishable by the composition of the essential oil from the flower buds. Although there are previous studies of the gastroprotective effect of the isolated constituents of E. viscosa, its infusions have not been investigated yet. AIM OF THE STUDY: The present study aimed to evaluate and compare the chemical composition and the gastroprotective effect of flower bud infusions of E. viscosa from chemotype A (EVCA) and chemotype B (EVCB). MATERIALS AND METHODS: Sixteen infusions were brewed with flower buds according to the traditional preparation mode and were analyzed through a UPLC-QTOF-MS/MS based metabolomic approach for determination of their metabolic fingerprints and quantification of bioactive compounds. Afterward, these data were analyzed by chemometric methods (OPLS-DA) for discrimination of the two chemotypes. Additionally, infusions of EVCA and EVCB (50, 100 and 200 mg/kg, p.o.) were evaluated on gastric ulcers induced by absolute ethanol (96%, 0.2 mL, p.o.) in mice. To elucidate the gastroprotective mechanisms, the effect of EVCA and EVCB on gastric acid secretion and gastric wall mucus was determined and the role of TRPV1 channels, prostaglandins, nitric oxide and KATP channels were assessed. Moreover, the oxidative stress-related parameters and the histological aspects of the stomach tissue were analyzed. RESULTS: The chemotypes can be discriminated from each other using UPLC-QTOF-MS/MS chemical fingerprints. Both chemotypes presented similar chemical compositions, consisting basically of caffeic acid derivatives, flavonoids and diterpenes. The quantification of bioactive compounds demonstrated that chemotype A possesses more ternatin, tanabalin and centipedic than chemotype B. EVCA and EVCB (50, 100 and 200 mg/kg, p.o.) significantly decreased the severity of ethanol-induced gastric lesions, as shown by a reduction in histological alterations and leucocyte infiltration in gastric tissue. The gastroprotective mechanism of both infusions involves an antioxidant effect, maintenance of gastric mucus and reduction gastric secretion. Stimulation of endogenous prostaglandins and nitric oxide release, activation of TRPV1 channels, and KATP channels are also involved in the gastroprotection of the infusions. CONCLUSION: The gastroprotective effect of EVCA and EVCB was equivalent and mediated through antioxidant and antisecretory actions, including the activation of TRPV1 receptors, stimulation of endogenous prostaglandins and nitric oxide, and opening of KATP channels. The presence of caffeic acid derivatives, flavonoids and diterpenes in both infusions is involved in mediating this protective effect. Our findings support the traditional use of infusions of E. viscosa for gastric disorders regardless of the chemotype.

Corn starch based films treated by dielectric barrier discharge plasma.

Cold plasma is an innovative strategy to strengthen the polysaccharide-based films characteristics. This study evaluated the effects of dielectric barrier discharge (DBD) plasma on the hydrophilic character, water vapor permeability (WVP), and tensile properties of corn starch-based films. Starch films were exposed to plasma processing operating at an excitation frequency of 200 Hz for 10, 15, and 20 min. DBD plasma resulted in further enhanced tensile strength and stiffness, and lower hydrophilicity and water solubility; however, it did not present significant effects on the WVP of the resulting films within the ranges studied. Higher hydrophobicity, strength, and stiffness were verified after 20 min. The results presented in this work suggest that the DBD plasma has the potential to make starch-based films a more suitable packaging material.

Chemical profile of colorful bean (Phaseolus vulgaris L) flours: Changes influenced by the cooking method.

This study aimed to determine how the cooking methods change the phenolics and saponins profiles, oligosaccharides, antinutrients and antioxidant properties of flours from colorful beans. The autoclave cooking consisted of: 6 h soaking and 5 min cooking (C5); and 20 min cooking without soaking (C20). Both cooking methods significantly promote changes on the chemical compounds studied, and the intensity of these variations were affected by the cultivars. Most of flours of C5 beans presented a lower loss of anthocyanins (3.9-70.0%), DPPH (11.7-87.2%), ABTS (0.0-82.7%), and tannins (0.0-90.0%) compared with C20. The cooked flours of Artico and Realce showed some similarities among chemical compounds, as well as the lowest concentration of tannins (0.0 mg‧g-1), antioxidant activity (0.40 µmol Trolox‧g-1), and higher amounts of oligosaccharides and acetylcholine. Most of cooked flours presented a reduction in phenolics and soyasaponins αg and ßg, and an increase in soyasaponins Ba and I and oligosaccharides (mainly C20 flours).

Volatile compounds associated to the loss of astringency in 'Rama Forte' persimmon fruit.

The volatile profile of 'Rama Forte' persimmon fruit treated with high concentration of CO2 or ethanol, as well as the association of specific volatile organic compounds (VOC) to the loss of astringency given by sensory and chemical evaluation were investigated in our study. Fruit were harvested at the maturity stage showing crisp texture and commercial color, and exposed to different astringency removal treatments: 70% CO2 for 18 h or 1.70 mL kg-1 ethanol for 6 h. Fruit were daily assessed for VOC, astringency, and soluble tannins content throughout eight days at room temperature. The HS-SPME/GC-MS analysis allowed to tentatively identify 31 volatile compounds and two fatty acids. A clear separation of the persimmon fruit from the beginning to the end of the post-treatment period was showed by PCA results, mainly from the fifth day for CO2-treated and the seventh day for ethanol-treated persimmon. The loadings from CO2 treatment highlighted the increase in the concentrations of a great number of compounds with the progression of the days. The compounds 2,4-di-tert-butylphenol and 1-pentadecanal were tentatively identified on fruit from both treatments after the longest periods and were absent on astringent non-treated fruit. These compounds are reported for the first time on persimmon fruit. Astringency tending to absent was noticed from the fifth and the seventh day after the fruit exposition to CO2 and ethanol vapor, respectively, when flesh soluble tannins concentrations close to 0.1% FW were found. Our study indicates a strong relationship between the astringency removal and the compounds 2,4-di-tert-butylphenol and 1-pentadecanal, supporting them as potential marker compounds for artificial deastringency in 'Rama Forte' persimmon fruit.

Transcriptome analysis of acerola fruit ripening: insights into ascorbate, ethylene, respiration, and softening metabolisms.

KEY MESSAGE: The first transcriptome coupled to metabolite analyses reveals major trends during acerola fruit ripening and shed lights on ascorbate, ethylene signalling, cellular respiration, sugar accumulation, and softening key regulatory genes. Acerola is a fast growing and ripening fruit that exhibits high amounts of ascorbate. During ripening, the fruit experience high respiratory rates leading to ascorbate depletion and a quickly fragile and perishable state. Despite its growing economic importance, understanding of its developmental metabolism remains obscure due to the absence of genomic and transcriptomic data. We performed an acerola transcriptome sequencing that generated over 600 million reads, 40,830 contigs, and provided the annotation of 25,298 unique transcripts. Overall, this study revealed the main metabolic changes that occur in the acerola ripening. This transcriptional profile linked to metabolite measurements, allowed us to focus on ascorbate, ethylene, respiration, sugar, and firmness, the major metabolism indicators for acerola quality. Our results suggest a cooperative role of several genes involved in AsA biosynthesis (PMM, GMP1 and 3, GME1 and 2, GGP1 and 2), translocation (NAT3, 4, 6 and 6-like) and recycling (MDHAR2 and DHAR1) pathways for AsA accumulation in unripe fruits. Moreover, the association of metabolites with transcript profiles provided a comprehensive understanding of ethylene signalling, respiration, sugar accumulation and softening of acerola, shedding light on promising key regulatory genes. Overall, this study provides a foundation for further examination of the functional significance of these genes to improve fruit quality traits.

UPLC-HRMS and NMR applied in the evaluation of solid-phase extraction methods as a rational strategy of dereplication of Phyllanthus spp. aiming at the discovery of cytotoxic metabolites.

The classical approach to drug discovery from natural products (NP's) requires strenuous and complex purification steps for the isolation and structural elucidation. Modern strategies as dereplication aim to accelerate the identification of known compounds present in a crude or partially purified extract. In this work, we investigated the influence of the solid-phase extraction (Oasis, Plexa, and Agilent C18 cartridges with and without organic modifiers) chemical profile obtained by UPLC-QTOF-MS and NMR and cytotoxicities of aqueous extracts from Phyllanthus niruri and P. amarus. Our results showed differences between the SPE cartridges and the mass recovered. P. niruri showed higher mass recovery than P. amarus indicating a higher amount of secondary metabolites. The UPLC-QTOF-MS analysis revealed that P. niruri crude extract presents higher contents of phenolic compounds than P. amarus. According to NMR analysis, P. niruri contained more tyrosine, corilagin, and glycosidic residues while P. amarus, presented higher content of ellagic acid. The different stationary phases, as well as mobile phases for exploratory SPE, enabled the exploitation of the different chemical functionalities within the Phyllanthus species. The SPE (MeOH:H2O 70:30 with C18 cartridges) samples showed greater in vitro cytotoxicity than the crude extracts, with IC50 ranging from 8.01 to 94.92 µg mL-1 against the tumor lines tested. The solid phase extraction allowed the concentration of molecules with desirable physicochemical characteristics, which might increase the hit of therapeutically useful substances.

Cashew apple fiber prevents high fat diet-induced obesity in mice: an NMR metabolomic evaluation.

Dietary fiber intake plays an important role in the prevention of obesity. This study aimed at investigating the effect of cashew fiber without low molecular weight compounds (CABwc) on obesity prevention and metabolomics in a murine model of diet-induced obesity. Mice were fed a chow diet (CD), a high-fat diet (HFD) or a high-fat diet supplemented with CABwc (10%) (HFD-CABwc) for 15 weeks. The body weight, abdominal fat, serum glucose levels, insulin and lipid profiles, satiety hormones such as leptin and ghrelin, digestive enzymes such as amylase and lipase, and inflammatory mediators such as TNF-α, IL-6, and adiponectin were measured, in addition to performing serum and hepatic tissue analyses. The metabolomic analysis was based on nuclear magnetic resonance (NMR) spectroscopy of serum and feces. The effects observed with ingestion of CABwc were appetite control and prevention of hyperglycemia, hyperinsulinemia and hypertriglyceridemia, as well as the prevention of the inflammatory process and reduction of liver injury caused by the HFD. In addition, NMR evidenced the presence of SCFAs in serum and feces of mice fed with HFD-CABwc. These findings suggest that CABwc promoted satiety in mice, improving the metabolism of glucose and lipids. Positive effects of obesity prevention may be associated with SCFA production.