Isolation and purification of chemical constituents of liquid culture of symbiotic Chaetomium globosum ML-4 of oyster was performed through silica gel column chromatography, gel filtration over Sephadex LH-20, preparative TLC and HPLC. Five compounds were obtained and their structures were determined as chaetoglobosin V(1), chaetoglobosin Vb(2), tyrosol(3), 5-methyluracil(4)and uracil(5), respectively, based on HR-MS and NMR data and comparison with literatures. In vitro cytotoxicity of compounds against human hepatocellular carcinoma cell line SMMC-7721 were measured byMTT method, and results showed that compound 1 could obviously inhibit the proliferation of SMMC-7721 cells with an IC₅₀ value of 60.5 mg•L⁻¹, while the IC₅₀ value of positive control cisplatin was 19.96 mg•L⁻¹. Further studies discovered that compound 1 could lead to G2 phase arrest in SMMC-7721 cells and induce SMMC-7721 cells apoptosis. The ratio of Bcl-2/Bax in SMMC-7721 cells was decreased. The expression of protein Caspases-3,-8,-9 was improved and the expression and phosphorylation level of Akt were reduced. Aforementioned results revealed that in vitro antitumor activity of compound 1 against SMMC-7721 cells were related to G2 phase cell cycle arrest and induced-apoptosis. The induced-apoptosis was involved in both the mitochondrial pathway and the death receptor pathway and connected with activity decline of PI3K/Akt signaling pathway.

Tyrosol and caffeic acid are biophenols that contribute to the beneficial properties of virgin olive oil. The influence of hydroxypropyl-β-cyclodextrin (HPβ-CD) on their respective antioxidant capacities was analyzed. The ORAC antioxidant activity of tyrosol (expressed as μM Trolox equivalents/μM Tyrosol) was 0.83 ± 0.03 and it increased up to 1.20 ± 0.11 in the presence of 0.8 mM HPβ-CD. However, the ORAC antioxidant activity of caffeic acid experienced no change. The different effect of HPβ-CD on each compound was discussed. In addition, the effect of increasing concentrations of different cyclodextrins in the development of ORAC-fluorescence (ORAC-FL) assays was studied. The ORAC signal was higher for HPβ-CD, followed by Mβ-CD, β-CD, γ-CD and finally α-CD. These results could be explained by the formation of inclusion complexes with fluorescein.

A capillary zone electrophoresis (CZE) method for the simultaneous determination of 20 polyphenols in wine was developed. The separation was performed using fused-silica capillaries of 75 μm i.d. and a 30 mM sodium tretraborate buffer solution at pH 9.2 with 5% isopropanol as a background electrolyte. A capillary voltage of +25 kV with pressure-assisted (3.5 kPa) separation from minute 18 was applied, thus achieving a total analysis time of <25 min. Instrumental quality parameters such as limits of detection (LOD, values between 0.3 and 2.6 mg/L), linearity (r(2)>> 0.990), and run-to-run and day-to-day precisions (RSD values lower than 6.5 and 15.7%, respectively) were established. Three different calibration procedures were evaluated for polyphenol quantitation in wines: external calibration using standards prepared in Milli-Q water, standard addition, and pseudomatrix-matched calibration using wine as a matrix. For a 95% confidence level, no statistical differences were observed, in general, between the three calibration methods (p values between 0.11 and 0.84), whereas for some specific polyphenols, such as cinnamic acid, syringic acid, and gallic acid, results were not comparable when external calibration was used. The CZE method using pseudomatrix-matched calibration was then proposed and applied to the analysis of polyphenols in 49 Spanish wines, showing satisfactory results and a wide compositional variation between wines. Electrophoretic profiles and other compositional data (e.g., peak areas of selected peaks) were considered as fingerprints of wines to be used for characterization and classification purposes. The corresponding data were analyzed by principal component analysis (PCA) to extract information on the most significant features contributing to wine discrimination according to their origins. Results showed that a reasonable distribution of wines depending on the elaboration areas was found, tyrosol and gallic, protocatechuic, p-coumaric, and caffeic acids being some representative discriminant compounds.

Candida krusei has been known to exhibit communal interactions such as pellicle formation and crawling out of nutritional broth. We noticed another possible interaction on agar surfaces, where C. krusei yeast cells formed mycelia along agar surfaces toward each other. We report here the results of experiments to study this interaction.

When C.krusei yeast cells are plated in parallel streaks, they form mycelia along agar surfaces toward other yeasts. They also detect the presence of Candida albicans and Candida glabrata across agar surfaces, while the latter two react neither to their own kind, nor to C. krusei. Secreted molecule(s) are likely involved as C.krusei does not react to heat killed C. krusei. Timing and rate of mycelia formation across distances suggests that mycelia start forming when a secreted molecule(s) on agar surface reaches a certain concentration. We detected farnesol, tyrosol and tryptophol molecules that may be involved with mycelial formation, on the agar surfaces between yeast streaks. Unexpectedly the amounts detected between streaks were significantly higher than would have expected from additive amounts of two streaks. All three Candida species secreted these molecules. When tested on agar surface however, none of these molecules individually or combined induced mycelia formation by C. krusei.

Our data confirms another communal interaction by C. krusei, manifested by formation of mycelia by yeast cells toward their own kind and other yeasts on agar surfaces. We detected secretion of farnesol, tyrosol and tryptophol by C. krusei but none of these molecules induced this activity on agar surface making it unlikely that they are the ones utilized by this yeast for this activity.

The olive pulp fraction contained in the residue generated in olive oil extraction by a two-step centrifugation process can be upgraded by using the cellulose fraction to produce ethanol and recovering high value phenols (tyrosol and hydroxytyrosol). Olive pulp was pretreated in a laboratory scale stirred autoclave at different temperatures (150-250 degrees C). Pretreatment was evaluated regarding cellulose recovery, enzymatic hydrolysis effectiveness, ethanol production by a simultaneous saccharification and fermentation process (SSF), and phenols recovery in the filtrate. The pretreatment of olive pulp using water at temperatures between 200 degrees C and 250 degrees C enhanced enzymatic hydrolysis. Maximum ethanol production (11.9 g/L) was obtained after pretreating pulp at 210 degrees C in a SSF fed-batch procedure. Maximum hydroxytyrosol recovery was obtained in the liquid fraction when pretreated at 230 degrees C.

OBJECTIVE

To assess the effect of tyrosol on the production of hydrolytic enzymes (by Candida biofilm cells) and acid (by Streptococcus mutans biofilms), as well as to quantify single and mixed biofilms of these species formed on acrylic resin (AR) and hydroxyapatite (HA).

METHODS

Candida and S. mutans biofilms were formed on AR and HA in the presence of tyrosol during 48 hours. Next, acid proteinase, phospholipase and hemolytic activities of Candida biofilm cells were determined, while acid production by S. mutans biofilms was assessed by pH determination. The effect of tyrosol on mature biofilms (96 hours) was evaluated through quantification of total biomass, metabolic activity, number of colony-forming units and composition of biofilms' extracellular matrix. Data were analyzed by one- and two-way ANOVA, followed by Tukey's and Holm-Sidak's tests (α = 0.05).

RESULTS

Treatments with tyrosol were not able to significantly reduce hydrolytic enzymes and acid production by Candida and S. mutans. Tyrosol only significantly reduced the metabolic activity of single biofilms of Candida species.

CONCLUSIONS

Tyrosol on its own had a limited efficacy against single and mixed-species oral biofilms. Its use as an alternative antimicrobial for topical therapies still demands more investigation.

There is limited knowledge regarding the impact of naturally occurring lipid-phenolic interactions on the biological properties of phenolics in virgin olive oil. Free and bound phenolics were isolated via sequential methanolic extraction at 30 and 60 °C, and were identified and quantified using reversed phase high performance liquid chromatography, liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), and gas chromatography. Decreased oleic acid concentrations and increased concentrations of palmitoleic acid, stearic, linoleic, and linolenic acids were observed in virgin olive oil after removal of free and bound lipid phenolic compounds. The presence of p-hydroxybenzoic acid and tyrosol bound to glycerides was determined via LC-MS/MS, which indicates natural lipid-phenolic interactions in virgin olive oil. Both free and lipid bound phenolic extracts exerted antiproliferative activities against the CRC1 and CRC5 colorectal cancer cell lines. The present work indicates that naturally occurring lipid-phenolic interactions can affect the biological properties of phenolics in virgin olive oil.

Characterization of phenolic compounds in olive oil has not been achieved as yet, owing to the complexities of their chemical structures and analytical matrix. The aim of this work is to optimize and validate a method for simultaneous separation and quantification of 13 phenolic compounds from extra-virgin olive oil: tyrosol, hydroxytyrosol, oleuropein glycoside, ferrulic acid, p-coumaric acid, cinnamic acid, p-hydroxybenzoic acid, gallic acid, caffeic acid, luteolin, apigenin, vanillic acid and 3,4-dihydroxybenzoic acid. A statistical central composite design, response surface analysis and the simultaneous optimization method of Derringer and Suich were used to separate all the peaks. These multivariate procedures were efficient in determining the optimal separation condition, using five peak-pair resolutions and runtime as responses. The optimized method employed a fused-silica capillary of 50 μm i.d.× 60 cm effective length with extended light path, 50 mmol L(-1) boric acid electrolyte, 10.2 pH, 25°C, injection of 50 mbar for 25s with application of reverse voltage (-30 kV for 5s) before setting the running voltage (+30 kV) with detection at 210 nm and a run time of 12 min. Peak resolutions are found to be very sensitive to pH values outside the 10.15-10.25 range but acceptable electropherograms can be obtained for a wide range of boric acid concentrations within this pH interval.

Many phenylalanine- and tyrosine-producing strains have used plasmid-based overexpression of pathway genes. The resulting strains achieved high titer and yield of phenylalanine and tyrosine. Chromosomally-engineered, plasmid-free producers have shown lower titer and yield than plasmid-based strains, but the former are advantageous in terms of cultivation cost and public health/environmental risk. Therefore, here, we engineered the Escherichia coli chromosome to create superior phenylalanine- and tyrosine-overproducing strains that did not depend on plasmid-based expression. Integration into the E. coli chromosome of two central metabolic pathway genes (ppsA and tktA) and eight shikimate pathway genes (aroA, aroB, aroC, aroD, aroE, aroG^fbr , aroL, and pheA^fbr ), controlled by the T7lac promoter, resulted in excellent titer and yield of phenylalanine; fbr indicates that the enzyme encoded by the gene was feedback-resistant. The generated strain could be changed to be a superior tyrosine-producing strain by substitution of pheA^fbr to tyrA^fbr A rational approach revealed that integration of seven genes (ppsA, tktA, aroA, aroB, aroC, aroG^fbr , and pheA^fbr ) was necessary as the minimum gene set for high-yield phenylalanine production in E. coli MG1655 (tyrR, adhE, ldhA, pykF, pflDC, and ascF deletant). The phenylalanine- and tyrosine-producing strains were further applied to generate phenyllactic acid-, 4-hydroxyphenyllactic acid-, tyramine-, and tyrosol-producing strains; yield of these aromatic compounds increased proportionally to the increase in phenylalanine and tyrosine yields.Importance Plasmid-free strains for aromatic compound production are desired in the aspect of industrial application. However, yield of phenylalanine and tyrosine have been considerably lower in plasmid-free strains than in plasmid-based strains. The significance of this research is that we succeeded to generate superior plasmid-free phenylalanine- and tyrosine-producing strains, by engineering E. coli chromosome, which was comparable to plasmid-based strains. The generated strains have a potential to generate superior strains for production of aromatic compounds. Actually, we demonstrated that 4 kinds of aromatic compounds could be produced from glucose with high yield (eg. 0.28 g-tyrosol/g-glucose).

Tyrosol is extensively used in the pharmaceutical industry as an important natural product from plants. In this study, an exogenous pathway involved in catalyzing tyrosine to tyrosol was introduced into Saccharomyces cerevisiae. Furthermore, The pyruvate decarboxylase gene pdc1 was deleted to redirect the flux distribution at the pyruvate node, and a bifunctional NAD⁺-dependent fused chorismate mutase/prephenate dehydrogenase from E. coli (EcTyrA) and its' tyrosine inhibition resistant mutant (EcTyrA^M53I/A354V) were heterologously expression in S. cerevisiae to tuning up the chorismate metabolism effectively directed the metabolic flux toward tyrosol production. Finally, the tyrosol yield of the engineered strain GFT-4 was improved to 126.74 ± 6.70 mg/g DCW at 48 h, increased 440 times compared with that of the control strain GFT-0 (0.28 ± 0.01 mg/g DCW). The new synergetic engineering strategy developed in this study can be further applied to increase the production of high value-added aromatic compounds derived from aromatic amino acid or shikimate in S. cerevisiae.

BACKGROUND

Olive oil is the major source of tyrosol which is a natural phenolic antioxidant. Olive oil constitutes a major component of the Mediterranean diet that is linked to a reduced incidence of chronic diseases.

OBJECTIVE

This study evaluates the effects of tyrosol on altered glycoprotein components in streptozotocin-induced diabetic rats.

METHODS

Diabetes mellitus was induced in male Wistar rats by streptozotocin (40 mg/kg body weight). These rats were administered tyrosol (20 mg/kg body weight) and glibenclamide (600 μg/kg body weight) orally daily for 45 days. Plasma glucose, plasma insulin, glycoprotein components such as hexose, hexosamine, sialic acid and fucose in the plasma, liver and kidney, and histopathogy of tissues were analyzed.

RESULTS

Diabetic rats revealed significant (p < 0.05) increase in the levels of glucose, hexose, hexosamine, sialic acid and fucose (277.17, 152.45, 100.43, 79.69 and 49.29 mg/dL) in the plasma; decrease in the levels of palsma insulin (6.12 μU/mL) and sialic acid (4.36 and 5.03 mg/g) in the liver and kidney; significant (p < 0.05) increase in hexose (49.33 and 46.82 mg/g), hexosamine (22.68 and 33.20 mg/g) and fucose (31.63 and 32.44 mg/g) in the liver and kidney. Further, periodic acid-Schiff staining of tissues revealed positive-stain accumulation in diabetic rats. Tyrosol treatment showed significant (p < 0.05) effects on all the biochemical parameters and histopathology studied in streptozotocin- nduced diabetic rats. Also, the in vitro study revealed the antioxidant effect of tyrosol.

CONCLUSIONS

Thus, tyrosol protects streptozotocin-induced diabetic rats from the altered glycoprotein components. Further, this study can be extrapolated to humans.

The neuroprotective effect of virgin olive oil (VOO) polyphenols has been related to their antioxidant effect. The main objective was to analyze how tyrosol and hydroxytyrosol contribute to the antioxidant and neuroprotective effects of VOO in a model of hypoxia-reoxygenation in rat brain slices. Rats were treated per os (po) (10 or 20 mg/kg/day) with hydroxytyrosol ethyl ether (HTEE), tyrosol ethyl ether (TEE), or 3,4-di-o-methylidene-hydroxytyrosol ethyl ether (MHTEE), used as a negative control for antioxidant effects. Lipid peroxidation was inhibited with HTEE, TEE, and MHTEE (from 5.0 ± 1.5 to 2.6 ± 1.5, 4.5 ± 1.5, and 4.8 ± 1.5 nmol/mg protein, respectively). However, all three compounds had similar neuroprotective effects: from 2.8 ± 0.07 to 1.8 ± 0.02 arbitrary units for HTEE, 1.4 ± 0.09 arbitrary units for TEE, and 1.3 ± 0.2 arbitrary units for MHTEE. All three compounds inhibited 3-nitrotyrosine production (from 3.7 ± 0.3 to 1.2 ± 0.03 nmol/0.1 g tissue for HTEE, 1.0 ± 0.2 nmol/0.1 g tissue for TEE, and 1.3 ± 0.1 nmol/0.1 g tissue for MHTEE), prostaglandin E2 production (from 55.7 ± 2.2 to 46.4 ± 1.9 pg/0.1 g tissue for HTEE, 24.7 ± 1.3 pg/0.1 g tissue for TEE, and 27.6 ± 2.6 pg/0.1 g tissue for MHTEE), whereas only HTEE inhibited IL1β production (from 35.7 ± 1.5 to 21.6 ± 0.8 pg/0.1 g tissue). Pearson correlation coefficients related neuroprotective effect with an antioxidant effect for HTEE (R = 0.72, p < 0.001), and inhibition of nitrosative stress (R = 0.78, 0.67, and 0.66 for HTEE, TEE, and MHTEE, respectively, p < 0.001) and inflammatory mediators (R = 0.72, 0.79, and 0.64 for HTEE, TEE, and MHTEE, respectively, p < 0.001) with all three compounds.

This work aims to analyze the chemical and biological evaluation of two extracts obtained by olive mill wastewater (OMW), an olive oil processing byproduct. The exploitation of OMW is becoming an important aspect of development of the sustainable olive oil industry. Here we chemically and biologically evaluated one liquid (L) and one solid (S) extract obtained by liquid-liquid extraction followed by acidic hydrolysis (LLAC). Chemical characterization of the two extracts indicated that S has higher phenol content than L. Hydroxytyrosol and tyrosol were the more abundant phenols in both OMW extracts, with hydroxytyrosol significantly higher in S as compared to L. Both extracts failed to induce cell death when challenged with endothelial cells and vascular smooth muscle cells in cell viability experiments. On the contrary, the higher extract dosages employed significantly affected cell metabolic activity, as indicated by the MTT tests. Their ability to counteract H₂O₂-induced oxidative stress and cell death was assessed to investigate potential antioxidant activities of the extracts. Fluorescence measurements obtained with the reactive oxygen species (ROS) probe H₂DCF-DA indicated strong antioxidant activity of the two OMW extracts in both cell models, as indicated by the inhibition of H₂O₂-induced ROS generation and the counteraction of the oxidative-induced cell death. Our results indicate LLAC-obtained OMW extracts as a safe and useful source of valuable compounds harboring antioxidant activity.

Keywords: antioxidant activity; olive mill wastewaters; phenolic extract; reactive oxygen species; vascular cells.

In this work, we investigated the interactions of human mesenchymal stem cells (hMSCs) with three-dimensional (3D) printed scaffolds displaying different scaffold architectures. Pressure-assisted microsyringe system was used to fabricate scaffolds with square (SQR), hexagonal (HEX), and octagonal (OCT) architectures defined by various degrees of curvatures. OCT represents the highest degree of curvature followed by HEX, and SQR is composed of linear struts without curvature. Scaffolds were fabricated from poly(L-lactic acid) and poly(tyrosol carbonate). We found that hMSCs attached and spread by taking the shape of the individual struts, exhibiting high aspect ratios (ARs) and mean cell area when cultured on OCT scaffolds as compared with those cultured on HEX and SQR scaffolds. In contrast, cells appeared bulkier with low AR on SQR scaffolds. These significant changes in cell morphology directly correlate with the stem cell lineage commitment, such that 80 ± 1% of the hMSCs grown on OCT scaffolds differentiated into osteogenic lineage, compared with 70 ± 4% and 62 ± 2% of those grown on HEX and SQR scaffolds, respectively. Cells on OCT scaffolds also showed 2.5 times more alkaline phosphatase activity compared with cells on SQR scaffolds. This study demonstrates the importance of scaffold design to direct stem cell differentiation, and aids in the development of novel 3D scaffolds for bone regeneration.

Collecting dietary intake data is associated with challenges due to the subjective nature of self-administered instruments. Biomarkers may objectively estimate the consumption of specific dietary items or help assess compliance in dietary intervention studies. Our aim was to use a panel of plasma and urine biomarkers to assess the validity of self-reported dietary intake using a modified Mediterranean Diet Scale (mMDS) among firefighters participating in Feeding America's Bravest (FAB), an MD cluster-randomized controlled trial. In our nested biomarker pilot study, participants were randomly selected from both the MD intervention group (n = 24) and the control group (n = 24) after 12-months of dietary intervention. At baseline data collection for the pilot study (t = 12-months of FAB), participants in the control group crossed-over to receive the MD intervention (active intervention) for 6-months. Participants in the intervention group continued in a self-sustained continuation phase (SSP) of the intervention. Food frequency questionnaires (FFQ), 13-item-mMDS questionnaires, 40 plasma fatty acids, inflammatory biomarkers and urinary hydroxytyrosol and tyrosol were analyzed at both time points. Spearman's correlation, t-tests and linear regression coefficients were calculated using SAS software. Overall, the mMDS derived from the FFQ was highly correlated with the specific 13-domain-mMDS (r = 0.74). The concordance between the two questionnaires for low and high adherence to MD was high for all the participants in the parent trial (κ = 0.76). After 6 months of intervention in the pilot study, plasma saturated fatty acid decreased in both groups (active intervention: -1.3 ± 1.7; p = 0.002; SSP: -1.12 ± 1.90; p = 0.014) and oleic acid improved in the SSP (p = 0.013). Intake of olive oil was positively associated with plasma omega-3 (p = 0.004) and negatively with TNF-α (p < 0.001) at baseline. Choosing olive oil as a type of fat was also associated with higher levels of plasma omega-3 (p = 0.019) at baseline and lower TNF-α (p = 0.023) at follow up. Intake of red and processed meats were associated with lower serum omega-3 (p = 0.04) and fish consumption was associated with lower IL-6 at baseline (p = 0.022). The overall mMDS was associated with an increase in plasma omega-3 (p = 0.021). Good correlation was found between nutrient intake from the FFQ and the corresponding plasma biomarkers (omega-3, EPA and DHA). In this MD randomized controlled trial, some key plasma biomarkers were significantly associated with key MD diet components and the overall mMDS supporting the validity of the mMDS questionnaire as well as compliance with the intervention.

In the present study, hybrid nanoflowers (HNFs) based on copper (II) or manganese (II) ions were prepared by a simple method and used as nanosupports for the development of effective nanobiocatalysts through the immobilization of lipase B from Pseudozymaantarctica. The hybrid nanobiocatalysts were characterized by various techniques including scanning electron microscopy (SEM), energy dispersion spectroscopy (EDS), X-ray diffraction (XRD), Raman spectroscopy, and Fourier transform infrared spectroscopy (FTIR). The effect of the addition of carbon-based nanomaterials, namely graphene oxide and carbon nanotubes, as well as magnetic nanoparticles such as maghemite, on the structure, catalytic activity, and operational stability of the hybrid nanobiocatalysts was also investigated. In all cases, the addition of nanomaterials during the preparation of HNFs increased the catalytic activity and the operational stability of the immobilized biocatalyst. Lipase-based magnetic nanoflowers were effectively applied for the synthesis of tyrosol esters in non-aqueous media, such as organic solvents, ionic liquids, and environmental friendly deep eutectic solvents. In such media, the immobilized lipase preserved almost 100% of its initial activity after eight successive catalytic cycles, indicating that these hybrid magnetic nanoflowers can be applied for the development of efficient nanobiocatalytic systems.

Phenylalkanols such as tyrosol and hydroxytyrosol (h-tyrosol), which possess antioxidant and anticancer properties, were phosphatidylated by phospholipase D (PLD)-catalyzed transphosphatidylation. After a 24-hour reaction of phosphatidylcholine (PC) and tyrosol with PLD, a new product was detected by TLC and identified to phosphatidyl-tyrosol by high-resolution MS and NMR analyses. The optimum reaction conditions were as follows: soyPC 50μmol, tyrosol 500μmol, ethyl acetate 1.6ml, PLD 1.6U, 0.2m sodium acetate buffer (pH 5.6) 0.8ml, 37°C for 24 hours. Under the optimum reaction conditions, the yields of phosphatidyl-tyrosol, hydroquinone (HQ), 2-(4-aminophenyl)ethanol (4APE), h-tyrosol and 2-phenylethanol (PEA) were 87±3.7, 13±1.3, 90±2.3, 64±5.5 and 85±1.0mol%, respectively. Furthermore, from the results of transphosphatidylation of soyPC with several phenylethanols and phenylpropanols, we established the following details about the reaction specificity of transphosphatidylation by PLD from Streptomyces sp.: (1) para-amino and para-hydroxyl groups in the benzene ring of PEA derivatives do not affect the transphosphatidylation by PLD, whereas meta-hydroxyl group slightly inhibits the transphosphatidylation. (2) Meta- and ortho-methyl groups in the benzene ring of PEA derivatives also slightly inhibit the transphosphatidylation. (3) Secondary and tertiary alcohols and hydroquinone are difficult to transphosphatidylate by PLD.

Water infusions of Ligustrum delavayanum and Ligustrum vulgare leaves and eight phenolics isolated therefrom have been assayed in vitro on ofloxacin-induced genotoxicity in the unicellular flagellate Euglena gracilis. The tested compounds luteolin, quercetin, luteolin-7-glucoside, luteolin-7-rutinoside, quercetin-3-rutinoside, apigenin-7-rutinoside, tyrosol and esculetin inhibited the mutagenic activity of ofloxacin (43 microM) in E. gracilis. Water infusions from leaves of L. delavayanum and L. vulgare showed higher antimutagenic effect (p(t) < 0.001). The activity of these samples against ofloxacin (86 microM)-induced genotoxicity was lower, but statistically significant (p(t) < 0.05), excluding the water infusion of L. delavayanum leaves (p(t) < 0.01). Efficacy of quercetin, luteolin-7-rutinoside, apigenin-7-rutinoside was insignificant. The antimutagenic effect of most phenolics we studied could be clearly ascribed to their DPPH scavenging activity, substitution patterns and lipophilicity.

Dysregulated receptor tyrosine kinase c-Met and its ligand HGF is valid and attractive molecular target for therapeutic blockade in cancer. Inspired by the chemical structure of the naturally occurring olive secoiridoid (-)-oleocanthal (1) and its documented anticancer activity against c-Met-dependent malignancies, a previous study reported tyrosol sinapate (4) as a c-Met inhibitor hit. This study reports additional semisynthetic optimization and SAR of 4 to improve its selective activity against c-Met-dependent breast cancer by increasing its capacity to inhibit c-Met phosphorylation. Forty-three compounds (5-47) were synthesized, among which the novel analog homovanillyl sinapate (HVS-16) was distinguished for its remarkable activity. HVS-16 substantially impaired c-Met-mediated proliferation, migration, and invasion across human breast cancer cell lines in two- and three-dimensional culture systems, while similar treatment doses were found to have effect neither on the non-tumorigenic human mammary epithelial cell growth nor on the c-Met independent breast cancer cell viability. HVS-16 showed a dose-dependent inhibition of ligand-mediated c-Met activation in human breast cancer cells. Docking studies revealed that HVS-16 fits very well inside c-Met crystal structures, satisfying critical interactions at the ATP binding site. This study identified important structural pharmacophoric features in HVS-16 and correlated its postulated binding pose with c-Met kinase assay data that would guide future olive secoiridoid bioisostere lead design. Results presented herein suggest HVS-16 as a promising c-Met inhibitor validated hit with potential to control invasive breast malignancies with aberrant c-Met activity.

OBJECTIVE

To study the chemical constituents from the roots of Rehmannia glutinosa.

METHODS

The compounds were isolated by various chromatographic methods and identified by spectroscopic analysis.

RESULTS

Twelve compounds were isolated and their structures were identified as 5-hydroxymethyl-pyrrole-2-carbaldehyde (1), 5-hydroxymethyl furfural (2), tyrosol (3), 5,6-dihydroxy-beta-ionone (4), 6-O-E-feruloyl ajugol (5), acteoside (6), leucosceptoside A (7), martynoside (8), isomartynoside (9), purpureaside C (10), jionoside A1 (11), and jionoside B1 (12).

CONCLUSIONS

Compounds 1, 3 and 9 were isolated from the genus Rehmannia for the first time.

Rhodiola plants are a valuable resource in traditional Chinese medicine. The objective of this study was to evaluate the correlation between ribosomal DNA internal transcribed spacer (ITS) sequences and the three active components in Rhodiola plants. For this, we determined ITS sequence polymorphisms and the concentrations of active components salidroside, tyrosol, and gallic acid in different Rhodiola species from the Tibetan Plateau. In a total of 23 Rhodiola samples, 16 different haplotypes were defined based on their ITS sequences. Analysis of the active components in these same samples revealed that salidroside was not detected in species with haplotypes H4, H5, or H10, tyrosol was not detected with haplotypes H3, H5, H7, H10, H14, or H15, and gallic acid was detected in with all haplotypes except H14 and H15. In addition, the concentrations of salidroside, tyrosol and gallic acid varied between samples with different haplotypes as well as those with the same haplotype, implying that no significant correlation exists between haplotype and salidroside, tyrosol or gallic acid concentrations. However, a statistically significant positive correlation was observed for among these three active components.

The autoregulatory d1 factor of the yeast, Saccharomyces cerevisiae, that induces the transition of vegetative cells into refractory resting forms, has been isolated from the cell-free culture medium as an individual crystalline compound. It has been shown to be 2-(4-hydroxyphenyl)ethane-1-ol which is also known as tyrosol. When added to the producer culture at 5-15 microM concentration, tyrosol stimulated the endogenous respiration of cells, but inhibited at 20-80 microM concentration. At 200-800 microM concentration, it induced the occurrence of resting forms. The action of tyrosol was not specific, for it also inhibited the cell respiration of the bacteria, Escherichia coli and Bacillus cereus, at 64-86 microM concentration.

A new isochromanone, named fraxitoxin, was isolated together with (-)-mellein and tyrosol from liquid cultures of Diplodia fraxini, a pathogen involved in the etiology of canker and dieback disease of Fraxinus spp. in Europe. It was characterized as 5-methoxy-3-methylisochroman-1-one using spectroscopic methods (essentially NMR and HR-EI-MS). Its absolute configuration (R) at C(3) was assigned by electronic circular dichroism (ECD) measurements and calculations. Phytotoxic activity of the compound was evaluated on ash, cork and holm oak leaves at concentration of 1 mg/ml by the leaf-puncture assay. Interestingly, fraxitoxin caused necrotic lesions only on ash leaves.

The innovative combination of ultrasound (Us) with a thermal exchanger to produce high quality extra virgin olive oil (EVOO) was studied using Nuclear Magnetic Resonance (NMR) spectroscopy and multivariate analysis (MVA). Major and minor metabolomic components of Apulian Coratina EVOO obtained using the two methods were compared. Early and late olive ripening stages were also considered. An increased amount of polyphenols was found for EVOOs obtained using the Us with respect to the conventional method for both early and late ripening stages (900.8 ± 10.3 and 571.9 ± 9.9 mg/kg versus 645.1 ± 9.3 and 440.8 ± 10.4 mg/kg). NMR spectroscopy showed a significant increase (P < 0.05) in polyunsaturated fatty acids (PUFA) as well as in the tyrosol and hydroxytyrosol derivatives, such as oleocanthal, oleacein, and elenolic acid, for both ripening stages. In conclusion, NMR spectroscopy provides information about the metabolomic components of EVOOs to producers, while the Us process increases the levels of healthy bioactive components.

Keywords: Coratina oil; Extravergin olive oil; NMR spectroscopy; Polyphenols; Ultrasound-assisted extraction system.