Antioxidant and anti-proliferative activity of free, conjugates and bound phenolic compounds from tomato and industrial tomato by-product.

The aim of this study was to evaluate the antioxidant and anti-proliferative activity of different fractions of phenolic compounds from tomato and tomato by-product. Soluble free phenolics and bound phenolics (BP) fractions from saladette tomato and industrial tomato by-product as well as, conjugated acid-hydrolysable phenolics (AHP) and alkaline-hydrolysable phenolics (AKHP) from saladette tomato, grape tomato and industrial tomato by-product, were tested. The scavenging effects of phenolic fractions on superoxide anion were determined using the PMS-NADH-NBT system. The myoglobin protection ratios of samples against hydroxyl radical were evaluated and the anti-proliferative effects in the MCF-7 breast cancer cell line were determined. AHP fraction from tomato by-product and AKHP fraction from grape and saladette tomato showed the highest inhibitory capacity of the superoxide anion (p ≤ 0.05). All samples had a myoglobin protection ratio from 30 to 50%. Regarding to cytotoxicity assays, all phenolic fractions from tomato by-product, as well as, BP, AHP, AKHP from saladette tomato and AHP from grape tomato showed important anti-proliferative activity against MCF-7 breast cancer cell line (IC50 < 20 µg/mL). From the results, we can conclude that tomato and tomato by-product are a good source of natural compound with important antioxidant and anti-proliferative activity.

Extraction of Lipophilic Antioxidants from Native Tomato Using Green Technologies.

Research background: Tomato (Solanum lycopersicum L.) fruit is highly consumed worldwide and contains high amounts of carotenoids and tocopherols, two powerful antioxidants. Native tomato genotypes are rarely used in large-scale market but serve as a reservoir to diversify the species gene pool and can be employed to obtain functional compounds. Extraction methods are currently changing towards cleaner procedures that are more efficient and environmentally friendly, including avoiding toxic or polluting solvents. Experimental approach: In this study, factorial and fractional factorial designs were used to evaluate the efficiency of digestive enzymes, sonication and green solvents to obtain lipophilic antioxidant extracts from native tomato. To monitor the efficiency of the extraction process, spectrophotometric quantification of total carotenoids and antioxidant activity was carried out, and then individual quantification of carotenoids and tocopherols in the extracts was done by HPLC. Results and conclusions: Digestive enzymes and sonication increased the carotenoid content and the antioxidant activity of the obtained extracts when applied individually. However, when these treatments were applied together and in combination with isopropyl acetate, a green solvent, the obtained extracts had the highest carotenoid and tocopherol contents as well as the maximal antioxidant activity. A correlation analysis suggested that antioxidant activity resulted from synergistic effects rather than individual compounds. Tomato extracts were obtained through a rapid and environmentally friendly extraction method and their antioxidant activity was enhanced. Novelty and scientific contribution: Tomato fruits have been the subject of numerous studies; however, functional compound extraction through environmentally friendly methods remains an attractive use of native tomato fruit, enhancing its limited production and harnessing a large amount of tomato product industry. There are few reports where environmentally friendly extraction methods are combined; even rarer are those where green solvents are also used. In this work, the combination of different environmentally friendly extraction methods improved the extraction of carotenoids and tocopherols and allowed to establish a more efficient process. These results could stimulate the use of clean technologies and make the native tomato more attractive for industrial or compound extraction processes.

Chemical Characterization and Evaluation of the Antihyperglycemic Effect of Lychee (Litchi chinensis Sonn.) cv. Brewster.

Lychee is a fruit of Asian origin with an exquisite flavor and an attractive reddish color. However, according to recent reports, the consumption of this fruit reduces the levels of blood glucose with adverse effects on human health such as encephalopathy and hypoglycemic. The objective of this work was to determine if the peel, pulp, and seed of "Brewster" lychee fruits harvested at two stages of maturity had antihyperglycemic effect. This effect was determined by an oral glucose tolerance test using Wistar rats. In addition, ultraviolet-visible spectrophotometry and high-resolution liquid chromatography were used to quantify phenolic compounds, flavonoids, organic acids (OAs), sugars, and antioxidant activity. Results indicated that stage I pulp (immature fruits) and stage II peel and seed (export mature fruits) reduced blood glucose levels, and the effects of the former two were synergistic with metformin. The pulp of mature fruits (stage II), however, lacked a hypoglycemic effect. Additionally, the peel and the seeds of these fruits presented a high antioxidant activity (as determined by DPPH [2,2-diphenyl-2-picryl-hydracyl] and ABTS+ [2,2-azino-bis-(3-ethylbenzothiazoline)-6-sulfonic acid] methods), which correlated well with the total content of phenolic compounds. The highest content of polyphenolics, flavonoids, and OAs was found in the extracts of the peel and seeds of both stages of maturity. It was therefore concluded that "Brewster" mature lychees are safe for human consumption, and both the seed and the peel can be useful sources for obtaining new compounds with antihyperglycemic activity.

Gene Cloning, Recombinant Expression, Characterization, and Molecular Modeling of the Glycolytic Enzyme Triosephosphate Isomerase from Fusarium oxysporum.

Triosephosphate isomerase (TPI) is a glycolysis enzyme, which catalyzes the reversible isomerization between dihydroxyactetone-3-phosphate (DHAP) and glyceraldehyde-3-phosphate (GAP). In pathogenic organisms, TPI is essential to obtain the energy used to survive and infect. Fusarium oxisporum (Fox) is a fungus of biotechnological importance due to its pathogenicity in different organisms, that is why the relevance of also biochemically analyzing its TPI, being the first report of its kind in a Fusarium. Moreover, the kinetic characteristics or structural determinants related to its function remain unknown. Here, the Tpi gene from F. oxysporum was isolated, cloned, and overexpressed. The recombinant protein named FoxTPI was purified (97% purity) showing a molecular mass of 27 kDa, with optimal activity at pH 8.0 and and temperature of 37 °C. The values obtained for Km and Vmax using the substrate GAP were 0.47 ± 0.1 mM, and 5331 µmol min-1 mg-1, respectively. Furthemore, a protein structural modeling showed that FoxTPI has the classical topology of TPIs conserved in other organisms, including the catalytic residues conserved in the active site (Lys12, His94 and Glu164). Finally, when FoxTPI was analyzed with inhibitors, it was found that one of them inhibits its activity, which gives us the perspective of future studies and its potential use against this pathogen.

Phytochemical Profile, Toxicity, and Pharmacological Potential of Peels from Four Species of Tropical Fruits.

Tropical fruit peels are generally discarded as waste, yet they contain bioactive substances that could have various uses; in addition, their pharmacological potential remains unexplored. This study aims to characterize the phytochemical profile, toxicity, and pharmacological potential of methanol extracts obtained from the peels of the following tropical fruit species: Annona squamosa L. (purple sugar apple), Annona reticulata L. (custard apple), Chrysophyllum cainito L. (green star apple), and Melicoccus bijugatus Jacq. (mamoncillo). Methanol peel extracts were obtained by maceration. All extracts contained flavonoids, anthraquinones, and triterpenoids as determined by colorimetric methods. A. squamosa and C. cainito exhibited the highest content of total phenols as assayed by the Folin-Ciocalteu method. M. bijugatus showed the highest content of total sugars (fructose, glucose, and sucrose) as determined by high-performance liquid chromatography. A. squamosa and C. cainito presented the highest antioxidant capacities (according to 2,2'-diphenyl-1-picrylhydrazyl, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid, and cupric reducing antioxidant capacity assays), displayed moderate toxicity against HCT-116 cells, and increased the vinblastine susceptibility of MCF-7/Vin+. A. squamosa and M. bijugatus extracts demonstrated modulation of acetylcholinesterase activity, whereas those of A. reticulata showed anti-inflammatory activity by inhibiting protein denaturation. These results confirm that tropical fruit peels can be valuable sources of bioactive compounds, and our findings provide new information about their pharmacologic potential so that they can be used as raw material for the development of new drugs aimed at treating a variety of ailments.

Antioxidant Capacity In Vitro and In Vivo of Various Ecotypes of Mexican Plum (Spondias purpurea L.).

Spondias purpurea L. is a fruit native to Mexico, however, it is found as far away as Brazil. It possesses a high commercial potential owing to its sensorial and nutritional qualities and its low cost of production. There exists a variety of ecotypes that have not been characterized and their adequate selection process, according to their strongest functional characteristics, will allow the establishment of improvement programs for this genetic resource. The object of this study was the chemical characterization and the determination of the in vitro and in vivo antioxidant capacity of 7 Spondias purpurea L. ecotypes. Differences were observed in the antioxidant capacity and the content of functional compounds among all the ecotypes analyzed. A high total phenolic content and a low flavonoid and carotenoid content were found, both in the epicarp and in the pulp. In each ecotype, the hydrophilic phase presented up to 40 times greater antioxidant capacity compared to the lipophilic phase. The hydrophilic phase of the epicarp of "Costeña Tierra Colorada" had the greatest antioxidant capacity and highest total phenolic content, whereas "Jocote" presented the lowest antioxidant capacity and total phenolic content. A positive correlation was observed between phenol levels and the antioxidant capacity in the epicarp. Regarding antioxidant activity in vivo, it was observed that in all analyzed concentrations of hydrophilic extracts of the epicarp of "Costeña Tierra Colorada" and in the highest "Jocote" concentrations, they provided thermo-protection against heat stress as well as a general well-being to the worm as evidenced by their high mobility.

Studies on phytochemical, antioxidant, anti-inflammatory, hypoglycaemic and antiproliferative activities of Echinacea purpurea and Echinacea angustifolia extracts.

CONTEXT: Echinacea (Asteraceae) is used because of its pharmacological properties. However, there are few studies that integrate phytochemical analyses with pharmacological effects. OBJECTIVE: Evaluate the chemical profile and biological activity of hydroalcoholic Echinacea extracts. MATERIALS AND METHODS: Density, dry matter, phenols (Folin-Ciocalteu method), flavonoids (AlCl3 method), alkylamides (GC-MS analysis), antioxidant capacity (DPPH and ABTS methods), antiproliferative effect (SRB assay), anti-inflammatory effect (paw oedema assay, 11 days/Wistar rats; 0.4 mL/kg) and hypoglycaemic effect (33 days/Wistar rats; 0.4 mL/kg) were determined in three Echinacea extracts which were labelled as A, B and C (A, roots of Echinacea purpurea L. Moench; B, roots, leaves, flowers and seeds of Echinacea purpurea; C, aerial parts and roots of Echinacea purpurea and roots of Echinacea angustifolia DC). RESULTS: Extract C showed higher density (0.97 g/mL), dry matter (0.23 g/mL), phenols (137.5 ± 2.3 mEAG/mL), flavonoids (0.62 ± 0.02 mEQ/mL), and caffeic acid (0.048 mg/L) compared to A and B. A, B presented 11 alkylamides, whereas C presented those 11 and three more. B decreased the oedema (40%) on day 2 similar to indomethacin. A and C showed hypoglycaemic activity similar to glibenclamide. Antiproliferative effect was only detected for C (IC50 270 µg/mL; 8171 µg/mL; 9338 µg/mL in HeLa, MCF-7, HCT-15, respectively). DISCUSSION AND CONCLUSION: The difference in the chemical and pharmacological properties among extracts highlights the need to consider strategies and policies for standardization of commercial herbal extracts in order to guarantee the safety and identity of this type of products.

Conformational and reactivity study of dithiophenyl-fucosyl ketals with theoretical chemical methods.

Carbohydrates can be used as substrates to synthesize new complex molecules; these molecules contain several chiral centers that can be used in organic synthesis. D-Fucose diphenyl thioacetal reacts differentially with acetone, and this paper describes a study of the mechanism of this reaction using theoretical chemistry methods. The conformer distribution was studied using a Monte Carlo method for the reaction products, and the obtained conformers were validated by calculating the hydrogen spin-spin coupling constants with the DFT/B3LYP/DGDZVP method. Results agreed with the experimental coupling constants with an adequate root mean squared deviation. The free energies and enthalpies of formation of the resulting global minimum conformers were calculated with the same method and with the thermochemical compound method CBS-4 M. This technique, combined with the conformational analysis, allowed comparison of the formation enthalpies of the compounds involved in this reaction, and, with this information, we can postulate the correct reaction pathway. Graphical abstract Reaction pathway.

Studies of (-)-pironetin binding to α-tubulin: conformation, docking, and molecular dynamics.

A comprehensive conformational analysis for the anticancer agent pironetin (1) was achieved by molecular modeling using density functional theory calculations at the B3PW91/DGTZVP level in combination with calculated and experimental (1)H-(1)H coupling constants comparison. Two solvent-dependent conformational families (L and M) were revealed for the optimum conformations. Docking studies of the pironetin-tubulin complex determined a quantitative model for the hydrogen-bond interactions of pironetin through the αAsn249, αAsn258, and αLys352 amino groups in α-tubulin, which supported the formation of a covalent adduct between the αLys352 and the ß carbon atom of the α,ß-unsaturated lactone. Saturation-transfer difference NMR spectroscopy confirmed that pironetin binds to tubulin, while molecular dynamics exposed a distortion of the tubulin secondary structure at the H8 and H10 α-helices as well as at the S9 ß-sheet, where αLys352 is located. A large structural perturbation in the M-loop geometry between the αIle274 and αLeu285 residues, an essential region for molecular recognition between α-α and ß-ß units of protofilaments, was also identified and provided a rationale for the pironetin inhibitory activity.

Structural reassignment, absolute configuration, and conformation of hypurticin, a highly flexible polyacyloxy-6-heptenyl-5,6-dihydro-2H-pyran-2-one.

The structural reassignment, absolute configuration, and conformational behavior of the highly flexible natural product hypurticin (pectinolide E), 6S-[3'S,5'R,6'S-triacetoxy-1Z-heptenyl]-5S-acetoxy-5,6-dihydro-2H-pyran-2-one (1), were ascertained by a molecular modeling protocol, which includes extensive conformational searching, geometry optimization by DFT B3LYP/DGDZVP calculations, and comparison between the theoretical (DFT) and experimental (1)H-(1)H NMR coupling constants. Hyptolide (2), a related cytotoxic 5,6-dihydro-2H-pyran-2-one that increased the S phase of the HeLa cell cycle, was employed as a reference substance to validate the theoretical protocol designed to characterize the 3D properties of compound 1. The related synthetic derivative, tri-O-acetyl-3,6-dideoxy-d-glucose diphenyldithioacetal (14), was prepared by a six-step reaction sequence starting from d-glucose and served as an enantiopure building block to reinforce the structural and configurational assignment of 1. This protocol proved to be an important tool for the structural characterization of highly flexible bioactive polyoxygenated natural products.