Tyrosol (TR), a major polyphenol found in extra virgin olive oil (EVOO), exerts several antioxidant effects. However, only scarce evidences are present regarding its activity on adipocytes and obesity. This study evaluated the role of TR in adipogenesis. Murine 3T3-L1 preadipocytes were incubated with TR (300 and 500 μM), and TR administration inhibited adipogenesis by downregulation of several adipogenic factors (leptin and aP2) and transcription factors (C/EBPα, PPARγ, SREBP1c, and Glut4) and by modulation of the histone deacetylase sirtuin 1. After complete differentiation, adipocytes treated with 300 and 500 μM TR showed a reduction of 20% and 30% in lipid droplets, respectively. Intracellular triglycerides were significantly reduced after TR treatment (p < 0.05). Mature adipocytes treated with TR at 300 and 500 μM showed a marked decrease in the inflammatory state and oxidative stress as shown by the modulation of specific biomarkers (TNF, IL6, ROS, and SOD2). TR treatment also acted on the early stage of differentiation by reducing cell proliferation (~40%) and inducing cell cycle arrest during Mitotic Expansion Clonal (first 48 h of differentiation), as shown by the increase in both S1 phase and p21 protein expression. We also showed that TR induced lipolysis by activating the AMPK-ATGL-HSL pathway. In conclusion, we provided evidence that TR reduces 3T3-L1 differentiation through downregulation of adipogenic proteins, inflammation, and oxidative stress. Moreover, TR may trigger adipose tissue browning throughout the induction of the AMPK-ATGL-UCP1 pathway and, subsequently, may have promise as a potential therapeutic agent for the treatment and prevention of obesity.

In this study, the untargeted phytochemical profile of non-edible parts of pomegranate (i.e., flowers, leaves and peels) was depicted by using untargeted metabolomics. Each sample was in vitro digested and then fermented simulating a large intestinal process, then monitoring the changes of phenolic bioaccessibility. The most abundant compounds in pomegranate extracts were polyphenols, terpenoids, sterols, alkaloids and amino acids. The untargeted metabolomic approach revealed a transformation of flavonoids over 24 h of in vitro fermentation. In particular, an increase in the % bioaccessibility for phenolic acids and tyrosols in flowers (probably due to the insoluble dietary fibre content, i.e., 27.1 g 100 g^-1) was observed. Accordingly, the highest % bioaccessibility was detected after 24 h of in vitro large intestine fermentation for flower (i.e., 53.8%). Results showed that non-edible parts of pomegranate could be used as ingredients for the development of potential value-added food and industrial products.

An endophytic fungus Pestalotiopsis microspora isolated from the fruits of Manilkara zapota was cultured in potato dextrose broth media. Chromatographic separation of the EtOAc extract of the broth and mycelium led to the isolation of a new azaphilonoid named pitholide E (1), in addition to previously identified pitholide B (2), pitholide D (3), pestalotin (LL-P880α) (4), PC-2 (5), LL-P880β (6), tyrosol (7) and 4-oxo-4H-pyran-3-acetic acid (8). An endophytic fungus P. microspora from M. zapota and the isolation of compounds 1-5, 7 and 8 from P. microspora are reported here for the first time.

BACKGROUND

The aim of the present work was to investigate the influence of fruit ripening on oil quality in an attempt to establish an optimum harvesting time for Chétoui olives, the second main olive variety cultivated in Tunisia.

RESULTS

Our results showed that many analytical parameters, i.e., peroxide value, UV absorbance at 232 and 270 nm, chlorophyll pigments, carotenoids and oleic acid contents decreased during ripening, whilst oil content and linolenic acid increased. Free acidity remained practically stable with a very slight rise at the highest maturity index. The trend of oxidative stability, total phenols and o-diphenols, showed an increase at the early stages followed by a reduction at more advanced stages of maturity. The major phenolic compounds, such as hydroxytyrosol, ligstroside aglycon, elenolic acid, acetoxy-pinoresinol and oleuropein aglycon, seemed to have the same behaviour. In the case of tyrosol, a strong decrease was observed directly related with the ripening progress.

CONCLUSIONS

On the basis of the evolution of the analytical parameters studied, the best stage of Chétoui olive fruits for oil processing seems to be at ripeness index higher than 2.0 and lower than 3.0.

Purpose: Several studies have demonstrated the properties of hydroxytyrosol, a phenolic compound present in olive oils and olives with a well-characterized impact on human health. Nevertheless, some knowledge gaps remain on its bioavailability and metabolism; overall concerning to the real rate per cent of absorption and biovailability of dietary hydroxytyrosol and the influence of the dietary food-containing hydroxytyrosol on it.

Methods: A double-blind study was performed including 20 volunteers who ingested 5 mg of hydroxytyrosol through diverse food matrices, to discover the influence on pharmacokinetics and bioavailability of HT metabolites (hydroxytyrosol acetate, 3,4-dihydroxyphenylacetic acid (DOPAC), tyrosol, and homovanillic alcohol) of the distinct matrices by UHPLC-ESI-QqQ-MS/MS.

Results: The HT pharmacokinetics after consumption of different food matrices was strongly dependent on the food matrix. In this aspect, the intake of extra virgin olive exhibited significantly higher plasma concentrations after 30 min of oral intake (3.79 ng/mL) relative to the control. Regarding the hydroxytyrosol bioavailability, the intake of extra virgin olive oil, as well as fortified refined olive, flax, and grapeseed oils provided significantly higher urinary contents (0.86, 0.63, 0.55, and 0.33 µg/mg creatinine, respectively) compared with basal urine, whereas hydroxytyrosol metabolites showed no significant changes. No differences were found between men and women.

Conclusions: The metabolic profile of hydroxytyrosol is influenced by the food matrix in which is incorporated, with the oily nature for the final bioavailability being relevant. Extra virgin olive oil was identified as the best matrix for this compound. The results described contribute to the understanding of the relevance of the food matrices for the final absorption of hydroxytyrosol and hence, the achievement of the highest health protection potential.

Keywords: Bioavailability; Food matrix; Human; Hydroxytyrosol metabolites; In vivo; Pharmacokinetic.

One new hydronaphthalenone derivative (1) was isolated from the broth extract of the endophytic fungus Daldinia eschscholtzii PSU-STD57 together with five known compounds, isosclerone (2), 8-methoxy-1-naphthol, 1,8-dimethoxynaphthalene, 2,6-dihydroxyphenyl-1-butan-1-one and tyrosol. The structures were assigned by spectroscopic methods. All the compounds were tested for antimicrobial activity against Staphylococcus aureus, methicillin-resistant S. aureus and Microsporum gypseum.

Potential metabolites of bioactive compounds are important for their biological activities and as authentic standards for metabolic studies. The phenolic compounds contained in olive oil are an important part of the human diet, and therefore their potential metabolites are of utmost interest. We developed a convenient, scalable, one-pot chemoenzymatic method using the arylsulfotransferase from Desulfitobacterium hafniense for the sulfation of the natural olive oil phenols tyrosol, hydroxytyrosol, and of their monoacetylated derivatives. Respective monosulfated (tentative) metabolites were fully structurally characterized using LC-MS, NMR, and HRMS. In addition, Folin-Ciocalteu reduction, 1,1-diphenyl-2-picrylhydrazyl radical scavenging, and antilipoperoxidant activity in rat liver microsomes damaged by tert-butylhydroperoxide were measured and compared to the parent compounds. As expected, the sulfation diminished the radical scavenging properties of the prepared compounds. These compounds will serve as authentic standards of phase II metabolites.

Red Globe grape polyphenol oxidase, partially purified using phase partitioning with Triton-X114, was used to study the oxidation of hydroxytytosol (HT) and its related compounds tyrosol (TS), tyrosol acetate (TSA), and hydroxytyrosol acetate (HTA). The enzyme showed activity toward both monophenols (monophenolase activity) and o-diphenols (diphenolase activity) with a pH optimum (pH 6.5) that was independent of the phenol used. However, the optimal temperature for diphenolase activity was substrate-dependent, with a broad optimum of 25-65 °C for HT, compared with the maximum obtained for HTA (40 °C). Monophenolase activity showed the typical lag period, which was modulated by pH, substrate and enzyme concentrations, and the presence of catalytic amounts of o-diphenols. When the catalytic power (Vmax/K(M)) was determined for both activities, higher values were observed for o-diphenols than for monophenols: 9-fold higher for the HT/TS pair and 4-fold higher for HTA/TSA pair. Surprisingly, this ratio was equally higher for TSA (2.2-fold) compared with that of TS, whereas no such effect was observed for o-diphenols. This higher efficiency of TSA could be related to its greater hydrophobicity. Acetyl modification of these phenols not only changes the kinetic parameters of the enzyme but also affects their antioxidant activity (ORAC-FL assays), which is lower in HTA than in HT.

BACKGROUND

Olive biophenols are emerging as a valued class of natural products finding practical application in the food, pharmaceutical, beverage, cosmetic and nutraceutical industries due to their powerful biological activity which includes antioxidant and antimicrobial properties. Olive mill waste water (OMWW), a by-product in olive oil manufacturing, is rich in biophenols such as hydroxytyrosol and tyrosol. The amount of biophenols depends on the cultivar, the geographical area of cultivation, and the seasonal conditions. The goal of this study was to develop a straightforward method to assess the economic value of OMWW via quantification of hydroxytyrosol and tyrosol.

RESULTS

The amount of hydroxytyrosol and tyrosol phenolic compounds in the OMWW from four different cultivars grown in four different regions of Sicily was analyzed using liquid-liquid and solid-liquid analytical protocols developed ad hoc. Results showed significant differences amongst the different cultivars and their geographical origin. In all samples, the concentration of hydroxytyrosol was generally from 2 to 10 times higher than that of tyrosol. In general, the liquid-liquid extraction protocol gave higher amounts of extracted biophenols. The cultivar Cerasuola had the highest amount of both hydroxytyrosol and tyrosol. The cultivar Nocellara Etnea had the lowest content of both biophenols.

CONCLUSIONS

A quick method to assess the economic value of olive mill waste water via quantification of hydroxytyrosol and tyrosol in olive phenolic enriched extracts is now available.

Acrylamide and phenolic compounds on both fresh and cooked olives were monitored by HPLC/MS-MS and reversed-phase-HPLC methods along different procedures: elaboration process, high hydrostatic pressure (HHP), cooking treatment and bioavailability evaluation. Acrylamide was not detected during the elaboration process and after HHP treatment. Hydroxytyrosol, tyrosol, oleuropein and verbascoside were the most important phenols after HHP treatment. The frying and baking processes on olives enhanced the formation of acrylamide and a significant reduction in the phenolic compounds. The frying process produced lower acrylamide concentration and less reduction of phenolic compounds than the baking process, while in the gastrointestinal digestion these compounds were slightly reduced if compared to the initial stage. As a conclusion, the best way to ingest high quantities of phenols and reduce acrylamide consumption is by ingesting the olives when they are fresh. In case the olives need to be cooked, specific time and temperature conditions shall be applied.

In a cytopathic effect inhibition assay, a standardized Rhodiola rosea root and rhizome extract, also known as roseroot extract (SHR-5), exerted distinct anti-influenza A virus activity against HK/68 (H3N2) (IC₅₀ of 2.8 µg/mL) without being cytotoxic. For fast and efficient isolation and identification of the extract's bioactive constituents, a high-performance countercurrent chromatographic separation method was developed. It resulted in a three-stage gradient elution program using a mobile phase solvent system composed of ethyl acetate/n-butanol/water (1 : 4 : 5 → 2 : 3 : 5 → 3 : 2 : 5) in the reversed-phase mode. The elaborated high-performance countercurrent chromatographic method allowed for fractionation of the complex roseroot extract in a single chromatographic step in a way that only one additional orthogonal isolation/purification step per fraction yielded 12 isolated constituents. They cover a broad polarity range and belong to different structural classes, namely, the phenylethanoid tyrosol and its glucoside salidroside, the cinnamyl alcohol glycosides rosavin, rosarin, and rosin as well as gallic acid, the cyanogenic glucoside lotaustralin, the monoterpene glucosides rosiridin and kenposide A, and the flavonoids tricin, tricin-5-O-β-D-glucopyranoside, and rhodiosin. The most promising anti-influenza activities were determined for rhodiosin, tricin, and tricin-5-O-β-D-glucopyranoside with IC₅₀ values of 7.9, 13, and 15 µM, respectively. The herein established high-performance countercurrent chromatographic protocol enables fast and scalable access to major as well as minor roseroot constituents. This is of particular relevance for extract standardization, quality control, and further in-depth pharmacological investigations of the metabolites of this popular traditional herbal remedy.

Olive mill wastewater (OMW) as a by-product of olive oil extraction process has significant polluting properties mainly related to high organic load, increased COD/BOD ratio, high phenolic content and relatively acidic pH. Raw OMW from Slovenian Istria olive oil mill and its polar fraction were investigated in this study. Chemical characterization of OMW polar fraction identified tyrosol as the most abundant phenolic product, followed by catechol. Lethal and sub-lethal effects of OMW matrix and its polar fraction were tested using a battery of bioassays with model organisms: bacteria Vibrio fischeri, algae Chlorella vulgaris, water fleas Daphnia magna, zebrafish Danio rerio embryos, clover Trifolium repens and wheat Triticum aestivum. Raw OMW sample was the most toxic to V. fischeri (EC₅₀ = 0.24% of OMW sample final concentration), followed by D. magna (EC₅₀ = 1.43%), C. vulgaris (EC₅₀ = 5.20%), D. rerio (EC₅₀ = 7.05%), seeds T. repens (EC₅₀ = 8.68%) and T. aestivum (EC₅₀ = 11.58%). Similar toxicity trend was observed during exposure to OMW polar fraction, showing EC₅₀ values 2.75-4.11 times lower comparing to raw OMW. Tested samples induced also sub-acute effects to clover and wheat (decreased roots, sprouts elongation); and to zebrafish embryos (increased mortality, higher abnormality rate, decreased hatching and pigmentation formation rate). A comprehensive approach using a battery of bioassays, like those used in this study should be applied during ecotoxicity monitoring of untreated and treated OMW.

The photocatalytic ozonation of a mixture of 3 phenols (gallic acid, tyrosol and syringic acid) has been conducted under different operating conditions. The individual adsorption of the phenol type compounds onto titanium dioxide (photocatalyst) has been first evaluated. Equilibrium conditions are attained in less than an hour while the isotherm curves reveal that adsorption intensity increases in order: syringic acid < tyrosol < gallic acid. When the photocatalytic ozonation is applied, an optimum in titanium dioxide concentration is experienced (1.5 g L(-1)). Direct comparison of the photocatalytic ozonation to other less sophisticated oxidation systems (i.e., single ozonation, catalytic ozonation, photo-ozonation, etc.) indicates a higher efficiency of the former in terms of ozone uptake.

Qin Pi (<i>Fraxinus chinensis</i> Roxb.) is commercially used in healthcare products for the improvement of intestinal function and gouty arthritis in many countries. Three new secoiridoid glucosides, (8<i>E</i>)-4''-<i>O</i>-methylligstroside (<b>1</b>), (8<i>E</i>)-4''-<i>O</i>-methyldemethylligstroside (<b>2</b>), and 3'',4''-di-<i>O</i>-methyl-demethyloleuropein (<b>3</b>), have been isolated from the stem bark of <i>Fraxinus chinensis</i>, together with 23 known compounds (<b>4</b>-<b>26</b>). The structures of the new compounds were established by spectroscopic analyses (1D, 2D NMR, IR, UV, and HRESIMS). Among the isolated compounds, (8<i>E</i>)-4''-<i>O</i>-methylligstroside (<b>1</b>), (8<i>E</i>)-4''-<i>O</i>-methyldemethylligstroside (<b>2</b>), 3'',4''-di-<i>O</i>-methyldemethyloleuropein (<b>3</b>), oleuropein (<b>6</b>), aesculetin (<b>9</b>), isoscopoletin (<b>11</b>), aesculetin dimethyl ester (<b>12</b>), fraxetin (<b>14</b>), <em>tyrosol</em> (<b>21</b>), 4-hydroxyphenethyl acetate (<b>22</b>), and (+)-pinoresinol (<b>24</b>) exhibited inhibition (IC₅₀ ≤ 7.65 μg/mL) of superoxide anion generation by human neutrophils in response to formyl-L-methionyl-L-leuckyl-L-phenylalanine/cytochalasin B (fMLP/CB). Compounds <b>1</b>, <b>9</b>, <b>11</b>, <b>14</b>, <b>21</b>, and <b>22</b> inhibited fMLP/CB-induced elastase release with IC₅₀ ≤ 3.23 μg/mL. In addition, compounds <b>2</b>, <b>9</b>, <b>11</b>, <b>14</b>, and <b>21</b> showed potent inhibition with IC₅₀ values ≤ 27.11 μM, against lipopolysaccharide (LPS)-induced nitric oxide (NO) generation. The well-known proinflammatory cytokines<i>,</i> tumor necrosis factor-alpha (TNF-α) and interleukin 6 (IL-6), were also inhibited by compounds <b>1</b>, <b>9</b>, and <b>14</b>. Compounds <b>1</b>, <b>9</b>, and <b>14</b> displayed an anti-inflammatory effect against NO, TNF-α, and IL-6 through the inhibition of activation of MAPKs and IκBα in LPS-activated macrophages. In addition, compounds <b>1</b>, <b>9</b>, and <b>14</b> stimulated anti-inflammatory M2 phenotype by elevating the expression of arginase 1 and Krüppel-like factor 4 (KLF4). The above results suggested that compounds <b>1</b>, <b>9</b>, and <b>14</b> could be considered as potential compounds for further development of NO production-targeted anti-inflammatory agents.

Keywords: Fraxinus chinensis; Oleaceae; anti-inflammatory activity; secoiridoid; stem bark.

Table olives contain a wide range of polyphenols responsible for protective effects on health that have been associated with a lower prevalence of chronic diseases. A new method to identify and quantify these compounds in table olives, by means of methanol:ethanol (1:1; v/v) extraction followed by LC-ESI-MS/MS, has been developed and validated. The chromatographic column Eclipse-XDB-C18, never used before in this kind of application, provided the best results using Milli-Q water with 0.025% acetic acid and acetonitrile with 5% acetone as eluents. This method allows the quantification of 17 polyphenols, namely, hydroxytyrosol, tyrosol, salidroside, hydroxytyrosol acetate, catechol, vanillic acid, caffeic acid, o-coumaric acid, p-coumaric acid, verbascoside; oleuropein; pinoresinol, apigenin, luteolin, luteolin-7-O-glucoside, quercetin and rutin. The new method has been validated and shows linear correlations (R²>0.996), recoveries superior to 95%, high sensitivity, adequate precision and accuracy (RSD < 15%) as well as a short chromatographic analysis of 9 min. Its application to the analysis of Marfil table olives enabled the quantification of 15 polyphenols, among which hydroxytyrosol (384.1 ± 81.2 mg/kg), tyrosol (201.2 ± 3.8 mg/kg), luteolin (88.0 ± 3.8 mg/kg) and salidroside (85.9 ± 3.2 mg/kg) stand out. Furthermore, this method allows to assess whether the intake of a certain number of olives can meet the health claim associated to olive oil polyphenols (Reg. EU n.432/2012). Our results indicate that the daily intake of only 7 olives, which corresponds to 8 g of edible portion, provide an amount of hydroxytyrosol and derivatives (e.g. oleuropein complex and tyrosol) of 5 mg, according to the health claim of the EU. In view of the results, it could be stated that table olives are an excellent source of bioactive compounds, thus emerging as a promising functional food.

This report describes the fabrication and application of a novel graphene/poly(urea-formaldehyde) composite modified electrode as a sensitive amperometric detector of CE. The composite electrode was fabricated on the basis of the in situ polycondensation of a mixture of graphenes and urea-formaldehyde prepolymers on the surface of a platinum disc electrode. It was coupled with CE for the separation and detection of salidroside and tyrosol in Rhodiola, a traditional Chinese medicine, to demonstrate its feasibility and performance. Salidroside and tyrosol have been well separated within 6 min in a 40 cm long capillary at a separation voltage of 12 kV using a 50 mM borate buffer (pH 9.8). The prepared graphene-based CE detector offered significantly lower detection potential, yielded enhanced signal-to-noise characteristics, and exhibited high resistance to surface fouling and enhanced stability. It showed long-term stability and reproducibility with relative standard deviations of less than 5% for the peak current (n = 15).

Food plays a central role in health, especially through consumption of plant-derived foods. Functional foods, supplements, and nutraceuticals are increasingly entering the market to respond to consumer demand for healthy products. They are foods, supplements, and ingredients which offer health benefits beyond the standard nutritional value. Some benefits are associated with phenolic compounds and phytochemicals with antioxidant properties. An olive pâté (OP) was added with antioxidants derived from olive mill wastewater (OMWW) to obtain a functional product rich in phenolic compounds. The olive pâté is produced from the ground olive pericarp, which shows an excellent natural antioxidant content. The OMWW is a waste product from oil processing, which is also rich in phenolic compounds. The result was a product rich in trans-resveratrol, OH tyrosol, and tyrosol in concentrations such as satisfying the European community's claims regarding the possible antioxidant action on plasma lipids with excellent shelf-life stability. The total phenolic content was assayed by a colorimetric method, the antioxidant activity by the ABTS [(2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)] test, the phenolic profile by Q Exactive Orbitrap LC-MS/MS. The shelf-life stability was confirmed by yeast, molds, and total microbial count, pH, and water activity determinations, and the best pasteurization parameters were determined. The palatability was judged as excellent.

Keywords: Olive Pâté; Q Exactive Orbitrap LC-MS/MS; antioxidants; nutraceutical; olive mill wastewater; olive oil.

Discovery of potential lead molecule is a challenging, and complex process which require lots of money, patience, and manpower. Human beings are using natural products, predominantly secondary metabolites, for this purpose since ancient time and they are still working on them as a potent source for drug discovery due to their wide structural diversity. Phenolic phytochemicals such as hydroxy<em>tyrosol</em> and <em>tyrosol</em> are natural antioxidant and involved in many biological disease cure. Herein, we have carried out the quantum chemical calculations for conformational analysis, geometry optimization and computation of electronic as well as optical properties of hydroxy<em>tyrosol</em> and its analogues (<b>1a-1k</b>) in terms of density functional theory by using Gaussian 09 program suite. The eventual charge transfer within the molecules has been confirmed by the analysis of frontier molecular orbitals. The molecular docking studies of <b>1a-1k</b> with cyclooxygenase-2 showed the noticeable binding affinity as compared to other nonsteroidal anti-inflammatory drugs <i>viz</i>. aspirin, naproxen and celecoxib. Computation of pharmacokinetics and pharmacological properties confirmed the lead/drug like potential of these screened molecules. Furthermore, the molecular dynamics simulation of best three docked ligands (<b>1f</b>, <b>1h</b> and <b>1k</b>)-receptor complex and their binding free energy calculations reveals that these molecules bind in the catalytic cavity of cyclooxygenase-2 and found stable during the 100 ns of simulation. Communicated by Ramaswamy H. Sarma.

Keywords: ADMET; Hydroxytyrosol; MM-PBSA; density functional theory; dynamics simulations.

Rhodiola is widely consumed in traditional folk medicine and nutraceuticals. To establish a procedure for the hydrogen (¹H)-NMR spectroscopic fingerprinting of secondary metabolites from three different Rhodiola species, the variation among three Rhodiola species were studied using ¹H-NMR metabolomics combined with multivariate data analysis. Gene expression programming (GEP) was used to generate a formula to distinguish Rhodiola crenulata from two other Rhodiola species. Finally, HPLC was used to demonstrate the results. Same metabolites were compared by quantitative ¹H-NMR (qNMR). Three Rhodiola species were clearly discriminated by ¹H-NMR fingerprinting involved 22 nuclear magnetic signals of chemical constituents. y = d₁₆₆ × 2 + C₁ + d₅₆ + d₂₃₆ - d₁₂₈ × C₂ can be used to distinguish R. crenulata from two other Rhodiola species by GEP. The gallic acid concentration in R. crenulata was significantly higher than in the other. Rhodiola species as was the level of salidroside. R. crenulata also exhibited substantially higher levels of α-glucose. The fatty acid level in Rhodiola kirilowii was lower than the other species. These findings demonstrated that ¹H-NMR fingerprinting combined with principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), hierarchical cluster analysis (HCA) and GEP can be used to distinguish different Rhodiola species and these methods were applicable and effective approaches for metabolic analysis, species differentiation, and quality assessment. In addition, gallic acid, salidroside, α-D-glucose, glycine, alanine, caffeic acid and tyrosol and are the discriminators.

Introduction: Pseudomonas aeruginosa is considered a dangerous pathogen, as it causes many human diseases, besides that it is resistant to almost all types of antibacterial agents. So, new strategies to overcome P. aeruginosa infection have evolved to attenuate its virulence factors and inhibit its quorum-sensing (QS) activity.

Purpose: This study investigated the effect of tyrosol and EDTA as anti-quorum-sensing and antivirulence agents against P. aeruginosa PAO1.

Methods: Anti-quorum activity of sub-minimum inhibitory concentrations (sub-MICs) of tyrosol and EDTA was tested using Chromobacterium violaceum (CV 12,472) biosensor bioassay. Miller assay was used to assess the inhibition of QS signal molecules by β-galactosidase activity determination. Also, their effects on the production of protease, lipase, lecithinase, and motility were tested. The inhibitory effects of these molecules on QS regulatory genes and exotoxins genes expression were evaluated by real-time PCR.

Results: Tyrosol and EDTA at sub-MICs inhibited the production of violacein pigment. Both compounds inhibited QS molecules production and their associated virulence factors (protease, lipase, lecithinase, and motility) (P≤ 0.05). Besides, the expression levels of QS regulatory genes (lasI, lasR, rhƖI, rhIR, pqsA, and pqsR) and exotoxins genes (exoS and exoY) were significantly reduced (P≤ 0.05).

Conclusion: Both tyrosol and EDTA can be used to fight P. aeruginosa infection as anti-quorum-sensing and antivirulence agents at their sub-MICs.

Keywords: C. violaceum; PAO1; QS regulatory genes; exotoxins; real-time PCR; signal molecules.

Rats were fed a high-fat diet and treated with acrylamide (21 mg/kg BW), harmane (7 mg/kg BW), N^Ɛ-(carboxymethyl)lysine (CML; 10 mg/kg BW), or the co-accumulation of the three chemicals to investigate the effects of harmful Maillard reaction products (MRPs) generated by heat processing of potatoes. Damage to the liver, kidney, gastrocnemius, and nervous system was confirmed by serum biochemical testing and histopathology examination after six weeks of feeding. Serum metabolomic analysis was performed using gas chromatography time-of-flight mass spectrometry (GC-TOF-MS). The metabolites detected by GC-TOF-MS were processed using chemometric techniques, and the results were evaluated using a t-test (p < 0.05) and calculated variable importance in projection (VIP) value (>1.5). Significant changes (increase or decrease) were identified in 13 metabolites, compared with the control. These metabolites were gluconolactone, tyrosol, 5-hydroxytryptophan, erythrose 4-phosphate, quinolinic acid, 2-ketobutyric acid, tyrosine, 3-hydroxy-3-methylglutaric acid, desaminotyrosine, L-cysteine, ribonic acid, cysteamine, and tryptophan. Among these, cysteamine and tryptophan are metabolites unique to the co-accumulation of all three test compounds in the treated rats. This study demonstrates that these harmful MRPs exert toxic effects on the liver, kidney, and nervous system via different metabolic pathways. Moreover, co-accumulation of these harmful MRPs caused differing toxic effects and changes in a variety of metabolites. These changes may contribute to the interaction of those harmful MRPs and also affect toxicity at the metabolic level.

Keywords: Acrylamide; Co-accumulation; Harmane; Metabolites; N(Ɛ)-(Carboxymethyl)lysine; Serum metabolomics.

Most of the yeast bypasses the developmental stage from simple unicellular yeast to elongated structure like hyphae. Regulation of this transition is governed by various quorum sensing and signalling molecules produced under different conditions of growth, that differ significantly, both physiologically and chemically. The evidence of fungal quorum sensing was uncovered ten years ago after the discovery of farnesol as first eukaryotic quorum sensing molecules in Candida albicans. In addition to farnesol, tyrosol was identified as second quorum sensing molecules in C. albicans controlling physiological activities. After the discovery of farnesol and tyrosol, regulation of morphogenesis through the production of chemical signalling molecules such as isoamyl alcohol, 2-phenylethyl alcohol, 1-dodecanol, E-nerolidol, etc. is reported in C. albicans. Some of the evidence suggests that the budding yeast Saccharomyces cerevisiae exhibits this type of regulation and the signals are regulated by aromatic alcohols which are the end product of amino acid metabolism. The effects of these molecules on morphogenesis are not similar in both yeasts, making comparisons hard. It is hypothesized that these signals works in microorganisms to derive a competitive advantage. Here, we present an example for utilization of competitive strategy by C. albicans and S. cerevisiae over other microorganisms.

Keywords: Candida albicans; Quorum sensing; alcohol dehydrogenase; biofilms; fusel alcohols; saccharomyces cerevisiae.

Hydroxytyrosol, a naturally occurred orthodiphenolic antioxidant molecule found in olive oil and olive mill wastewaters, was obtained from the wet hydrogen peroxide photocatalytic oxidation of its monophenolic precursor tyrosol. The liquid-phase oxidation of tyrosol to hydroxytyrosol was performed by use of an iron-containing heterogeneous catalyst (Al-Fe)PILC with the assistance of UV irradiation at 254 nm and at room temperature. The spectroscopic and HPLC data of the synthesized compound proved to coincide fully with those of a pure sample obtained by continuous countercurrent extraction. This reaction was found to be light-induced. The hydroxytyrosol synthesis reaction reached its maximum yield of 64.36% under the optimized operating conditions of 3.6 mM tyrosol, 0.5 g L(-1) catalyst, and 10(-2) M H2O2 with the assistance of UV light. Increasing the initial hydrogen peroxide concentration more than 10(-2) M has a diminishing return on the reaction efficiency. Catalyst can be recuperated by means of filtration and then reused in a next run after regeneration since its activity did not significantly decrease (<10%). The reaction synthesis is operationally simple and could find application for industrial purposes.