Context: Rhodiola crenulata (Hook. f. et Thoms.) H. Ohba (Crassulaceae) is used to prevent and treat acute mountain sickness. However, the mechanisms underlying its effects on the central nervous system remain unclear.

Objective: To investigate the effect of Rhodiola crenulata on cellular metabolism in the central nervous system.

Materials and methods: The viability and Hif-1α levels of microglia and neurons at 5% O₂ for 1, 3, 5 and 24 h were examined. We performed the binding of salidroside (Sal), rhodiosin, tyrosol and p-hydroxybenzyl alcohol to Hif-1α, Hif-1α, lactate, oxidative phosphorylation and glycolysis assays. Forty male C57BL/6J mice were divided into control and Sal (25, 50 and 100 mg/kg) groups to measure the levels of Hif-1α and lactate.

Results: Microglia sensed low oxygen levels earlier than neurons, accompanied by elevated expression of Hif-1α protein. Salidroside, rhodiosin, tyrosol, and p-hydroxybenzyl alcohol decreased BV-2 (IC₅₀=1.93 ± 0.34 mM, 959.74 ± 10.24 μM, 7.47 ± 1.03 and 8.42 ± 1.63 mM) and PC-12 (IC₅₀=6.89 ± 0.57 mM, 159.28 ± 8.89 μM, 8.65 ± 1.20 and 8.64 ± 1.42 mM) viability. They (10 μM) reduced Hif-1α degradation in BV-2 (3.7-, 2.5-, 2.9- and 2.5-fold) and PC-12 cells (2.8-, 2.8-, 2.3- and 2.0-fold) under normoxia. Salidroside increased glycolytic capacity but attenuated oxidative phosphorylation. Salidroside (50 and 100 mg/kg) treatment increased the protein expression of Hif-1α and the release of lactate in the brain tissue of mice.

Conclusions: These results suggest that Sal induces metabolic reprogramming by regulating the Hif-1α signalling pathway to activate compensatory responses, which may be the core mechanism underlying the effect of Rhodiola crenulata on the central nervous system.

Keywords: Rhodiola crenulata; glycolysis; lactate; oxidative phosphorylation.

Biofilm formation conferring pathogenicity is a survival strategy for Pseudomonas aeruginosa. P. aeruginosa's virulence may differ due to differences in host-microbe interactions and the growth environment. The epithelial cell line within the respiratory system and the keratinocytes on the skin form the first physical barrier of defence. P. aeruginosa spp. biofilm formation and virulence factor secretion with and without quorum quenching (QQ) treatment was studied in co-culture using A549 and HaCaT cell lines; pyocyanin and rhamnolipid productions and elastolytic activity as virulence factors were quantified by independent assays. Biofilm formation was evaluated under dynamic conditions by quantifying total carbohydrates, alginate, proteins and eDNA. A sandwich ELISA was performed to study IL-8 secretion by the epithelial cells. The difference in gene expression of the quorum sensing (QS) and virulence factors between strains during individual and combination treatments was analysed by qPCR. Combination treatment by farnesol and tyrosol was more effective against P. aeruginosa biofilms when grown in co-cultures. The strain RBHi was found to be 3 to 4 times more virulent compared to PAO1 and NCTC 10,662, respectively, and combination treatment was more effective against RBHi strain when grown in co-culture with A549 cell line. The addition of quorum quenchers (QQs) individually and in combination reduced IL-8 secretion by A549 cells. Relative mRNA expression showed upregulation of the QS genes and virulence factors. Co-culture of P. aeruginosa and HaCaT cell line showed a general decrease in gene expression, especially in the case of P. aeruginosa RBHi when treated with farnesol and tyrosol combination.Key points• Differentiating the interactions of biofilm formed by different phenotypes of P. aeruginosa, NCTC 10,662 (non-mucoid), PAO1 (semi mucoid) and RBHi (heavily mucoid).• Biofilm formed by these P. aeruginosa strains on two commonly afflicted tissues represented by A549 (lung) and HaCaT (skin) cell lines.• Anti-biofilm/anti-virulence roles of quorum quenchers, tyrosol and farnesol in co-cultures.

Keywords: Acyl-homoserine lactones; Biofilms; Farnesol; Pseudomonas aeruginosa; Quorum quenchers; Tyrosol.

Tyrosol (Ty) and hydroxytyrosol (HTy) are valuable dietary phenolic compounds present in olive oil and wine, widely used for food, nutraceutical and cosmetic applications. Ty and HTy are endowed with a number of health-related biological activities, including antioxidant, antimicrobial and anti-inflammatory properties. In this work, we developed a sustainable, biocatalyzed flow protocol for the chemo- and regio-selective oxidation of Ty into HTy catalyzed by free tyrosinase from Agaricus bisporus in a gas/liquid biphasic system. The aqueous flow stream was then in-line extracted to recirculate the water medium containing the biocatalyst and the excess ascorbic acid, thus improving the cost-efficiency of the process and creating a self-sufficient closed-loop system. The organic layer was purified in-line through a catch-and-release procedure using supported boronic acid that was able to trap HTy and leave the unreacted Ty in solution. Moreover, the acetate derivatives (TyAc and HTyAc) were produced by exploiting a bioreactor packed with an immobilized acyltransferase from Mycobacterium smegmatis (MsAcT), able to selectively act on the primary alcohol. Under optimized conditions, high-value HTy was obtained in 75% yield, whereas TyAc and HTyAc were isolated in yields of up to 80% in only 10 min of residence time.

Keywords: acetate derivatives; biocatalysis; flow chemistry; hydroxytyrosol; oxidation; tyrosol.

Three-dimensional (3D) printing has emerged as a valuable tool in medicine over the past few decades. With a growing number of applications using this advanced processing technique, new polymer libraries with varied properties are required. Herein, we investigate tyrosol-based poly(ester-arylate)s as biodegradable inks in fused deposition modeling (FDM). Tyrosol-based polycarbonates and polyesters have proven to be useful biomaterials due to their excellent tunability, nonacidic degradation components, and the ability to be functionalized. Polymers are synthesized by polycondensation between a custom diphenol and commercially available diacids. Thermal properties, degradation rates, and mechanical properties are all tunable based on the diphenol and diacid chosen. Evaluation of material print as it relates to chemical structure, molecular weight, and thermal properties was explored. Higher-molecular-weight polymers greater than 50 kDa exhibit thermal degradation during printing and at some points are too viscous to print. It was determined that polymers with lower processing temperatures and molecular weights were printable regardless of the structure. An exception to this was pHTy6 that was printed at 65 kDa with minimal degradation. This is most likely due to its low melting temperature and, as a result, lower printing temperatures. Additionally, chemical improvements were made to incorporate thiol-alkene click chemistry as a means for postprint curing. Low-molecular-weight pHTy6 was end-capped with alkene functionality. This material was then formulated with either a dithiol for chain extension or tetrathiol for cross-linking. Scaffolds were cured after printing for 5, 15, 30 and 60 min intervals where longer cure times resulted in a tougher material. This design builds on the library of biologically active materials previously explored and aims to bring new biomaterials to the field of 3D-printed personal medicine.

Keywords: 3D printing; biodegradable; direct ink writing; polymer; thiol−ene click chemistry; tyrosol.

This study was dedicated to the optimization and preparation of chitosan-coated liposomes (chitosomes) as promising nanocarriers for retention of olive leaf extract optimized by Response surface methodology (RSM) based on central composite design. Accordingly, the best sample was chosen for further tests with the encapsulation efficiency, stability and electrical conductivity of 94%, 98% and 9.545 mS respectively. The average size of the optimal chitosome and nanoliposome were lower than 100 nm and the zeta potential was altered from a negative charge to positive after addition coating process with chitosan. Moreover, the differential scanning calorimetry of blank and loaded chitosome revealed the increase of fluidity and lower temperature of phase transition in loaded chitosome compared to blank one. FTIR spectra demonstrated that electrostatic interactions and hydrogen bonds occur between phospholipid polar groups, chitosan amine moieties and major olive leaf extract polyphenols including oleuropein and hydroxy tyrosol. Furthermore, the optimal loaded chitosome had the highest stability during 25 days at the temperature of 4 °C. Finally, the in vitro release tests were best fitted with Peppas-Sahlin and Kopcha models in food simulants and gastrointestinal simulated juice respectively revealing erosion-based release model.

Supplementary information: The online version contains supplementary material available at (10.1007/s13197-021-04972-2).

Keywords: Chitosome; Olive leaf extract; Optimization; Release; Response surface methodology.

Bacterial tyrosinases, as in the case of other bacterial oxidative enzymes, have been found to possess biochemical characteristics that typically make them more suited to applications requiring special operational conditions such as alkaline pH, high or low temperature, the presence of organic solvents, and the presence of inhibitors. Even though a great deal is known about fungal tyrosinases, bacterial tyrosinases still vastly remain underexplored for their potential application in organic synthesis. A literature survey in particular highlights the gaps in our knowledge pertaining to their biochemical properties. Bacterial tyrosinases have not only shown promise in the synthesis of medically important compounds such as L-3,4-dihydroxyphenylalanine (L-DOPA) and melanin but have also seen application in cross-linking reactions of proteins and the polymerization of environmental pollutants. Their ability to catalyse o-hydroxylation reactions have shown some degree of promise in the biocatalytic conversion of resveratrol to piceatannol, tyrosol to hydroxytyrosol, and many more. In this review, we will explore the world of bacterial tyrosinases, their current applications, and future perspectives for the application of these enzymes in organic synthesis.

Keywords: Bacteria; Cross-linking; L-DOPA; Melanin; Organic synthesis; Tyrosinase.

<i>Biscogniauxia rosacearum,</i> recognized for the first time as a pathogen involved in grapevine trunk diseases in Paveh (west of Iran) vineyards, produced <i>meso</i>-2,3-butanediol (<b>1</b>) as the only phytotoxin. Nectriapyrone (<b>2</b>), (3<i>R</i>)-5-methylmellein (<b>3</b>), (3<i>R</i>)-5-methyl-6-methoxymellein (<b>4</b>), and <em>tyrosol</em> (<b>5</b>) were instead produced as phytotoxins from a strain of the same fungus isolated from oak trees in Zagros forests of Gilan-e Gharb, Kermanshah Province. They were identified comparing their ¹H and ¹³C NMR, ESIMS, and specific optical rotation data with those already reported in the literature. The phytotoxicity of metabolites (<b>1-5</b>) was estimated by leaf puncture assay on <i>Quercus ilex</i> L. and <i>Hedera helix</i> L., and by leaf absorption assay on grapevine (<i>Vitis vinifera</i> L.) at a concentration of 5 × 10^-3 and 10^-3 M. Tested on grapevine, <i>meso</i>-2,3-butanediol (<b>1</b>) and (3<i>R</i>)-5-methyl-6-methoxymellein (<b>4</b>) resulted to be the most phytotoxic compounds. On <i>Q. ilex</i>, nectriapyrone (<b>2</b>) and <em>tyrosol</em> (<b>5</b>) showed severe necrosis at the highest concentration while none of the compounds (<b>1</b>-<b>5</b>) was active on <i>H. helix</i>. Furthermore, the phytotoxicity of compounds <b>3</b> and <b>4</b> was also compared with that of some related natural melleins to perform a structure-activity relationship (SAR) study. The results of this study were also discussed.

Keywords: Biscogniauxia rosacearum; SAR studies; grapevine; isocoumarines; oak trees; phytotoxins.

Phenolic compounds constitute an important family of natural bioactive compounds responsible for the medicinal properties attributed to Bryophyllum plants (genus Kalanchoe, Crassulaceae), but their production by these medicinal plants has not been characterized to date. In this work, a combinatorial approach including plant tissue culture, untargeted metabolomics, and machine learning is proposed to unravel the critical factors behind the biosynthesis of phenolic compounds in these species. The untargeted metabolomics revealed 485 annotated compounds that were produced by three Bryophyllum species cultured in vitro in a genotype and organ-dependent manner. Neurofuzzy logic (NFL) predictive models assessed the significant influence of genotypes and organs and identified the key nutrients from culture media formulations involved in phenolic compound biosynthesis. Sulfate played a critical role in tyrosol and lignan biosynthesis, copper in phenolic acid biosynthesis, calcium in stilbene biosynthesis, and magnesium in flavanol biosynthesis. Flavonol and anthocyanin biosynthesis was not significantly affected by mineral components. As a result, a predictive biosynthetic model for all the Bryophyllum genotypes was proposed. The combination of untargeted metabolomics with machine learning provided a robust approach to achieve the phytochemical characterization of the previously unexplored species belonging to the Bryophyllum subgenus, facilitating their biotechnological exploitation as a promising source of bioactive compounds.

Keywords: Kalanchoe; artificial intelligence; bioactive compounds; mineral nutrition; phytochemistry; plant biotechnology; plant tissue culture; polyphenols; secondary metabolism.

Ceramides are a class of sphingolipids which have recently been shown to be better cardiovascular disease (CVD) risk predictors than traditional CVD risk biomarkers. Tyrosol (TYR) is a dietary phenolic compound known to possess cardioprotective effects per se or through its in vivo active metabolite hydroxytyrosol. The purpose of this study was to evaluate the effects of the co-administration of white wine (WW) and TYR on circulating levels of ceramides and other lipids in humans at high CVD risk. Volunteers underwent a randomized controlled crossover clinical trial (4-week duration per intervention) with three different interventions: control, WW, and WW enriched with a capsule of TYR (WW + TYR). Endothelial function cardiovascular biomarkers and plasma lipidomic profile were assessed before and after each intervention. It was found that the WW + TYR intervention resulted in lower levels of three ceramide ratios, associated with an improvement of endothelial function (Cer C16:0/Cer C24:0, Cer C18:0/Cer C24:0, and Cer C24:1/Cer C24:0), when compared to the control intervention. Moreover, WW + TYR was able to minimize the alterations in plasma diacylglycerols concentrations observed following WW. Overall, the results obtained show that the antioxidant TYR administered with WW exerts beneficial effects at the cardiovascular level, in part by modulating blood lipid profile.

Keywords: cardiovascular disease; ceramides; ethanol; randomized clinical trial; tyrosol; white wine.

Heavy metals act as co-factors for several microbial enzymes and are required in low concentrations for proper biological functioning of yeasts. Since concentrations beyond permitted threshold can damage cell's functionality and viability, metal tolerance in yeasts towards such heavy metals is therefore desirable during fermentation. Tyrosol, a quorum sensing molecule in yeasts, protects yeasts from oxidative stress induced by various factors, but the performance of the molecule under heavy metal induced stress is not known. In this investigation, metal tolerance of four species of endemic yeasts from northeast India, viz Wickerhamomyces anomalus, Candida tropicalis, Saccharomyces cerevisiae and Candida glabrata, isolated from traditional starter culture cakes were tested towards zinc (Zn+2), manganese (Mn+2), cobalt (Co+2) and copper (Cu+2) in presence and absence of tyrosols retrieved from these isolates. The decreasing order of tolerance of isolates were found to be Mn+2 > Zn+2 > Co+2 > Cu+2. Under the influence of tyrosols, isolates showed enhanced growth in their upper metal tolerance limit. C. tropicalis showed enhanced growth (2-48 fold, p<0.0001) in all the tested metal consisting medium (2 mM Zn+2, 5 mM Mn+2, 2 mM Co+2 and 1 mM Cu+2) whilst W. anomalus, C. glabrata, S. cerevisiae showed increased growth (3-17 fold, p<0.0001) in Zn+2 (2 mM), Mn+2 (5 mM) and Cu+2 (1 mM) augmented medium. Overall result suggests that tyrosol exert protective effect under heavy metal induced stress, which could be useful in enhancing the quality of fermented products.

Keywords: Yeasts; cobalt (Co); copper (Cu); heavy metals; manganese (Mn); tyrosol; zinc (Zn).

Microglia are the main immune cells in the brain playing a critical role in neuroinflammation, and numerous pieces of evidence have proved that energy metabolism is closely associated with inflammation in activated microglia. Salidroside (Sal) isolated from Tibetan medicine Rhodiola crenulate can inhibit microglial hypoxia inflammation (HI). However, whether the inhibition is due to the intervening energy metabolic process in microglia is not clear. In this work, the hypoxic microenvironment of BV2 microglial cells was simulated using deferoxamine (DFO) in vitro and the change of cell metabolites (lactate, succinate, malate, and fumarate) was real-time online investigated based on a cell microfluidic chip-mass spectrometry (CM-MS) system. Meanwhile, for confirming the metabolic mechanism of BV2 cells under hypoxia, the level of HI-related factors (LDH, ROS, HIF-1α, NF-κB p65, TNF-α, IL-1β, and IL-6) was detected by molecular biotechnology. Integration of the detected results revealed that DFO-induced BV2 cell HI was associated with the process of energy metabolism, in which cell energy metabolism changed from oxidative phosphorylation to glycolysis. Furthermore, administration of Sal treatment could effectively invert this change, and two metabolites of Sal were identified: tyrosol and 4-hydroxyphenylacetic acid. In general, we illustrated a new mechanism of Sal for reducing BV2 cell HI injury and presented a novel analysis strategy that opened a way for real-time online monitoring of the energy metabolic mechanism of the effect of drugs on cells and further provided a superior strategy to screen natural drug candidates for HI-related brain disease treatment.

Keywords: hypoxia inflammation; metabolic mechanism; microfluidic chip-mass spectrometry; microglia; salidroside.

Phytochemical investigation on the concentrate of Huangjing wine, resulted in the isolation of three new <em>tyrosol</em> derivatives 4<i>'''</i>-hydroxyphenethyl 2-(R)-hydroxy-3-phenylpropionate (<b>1</b>), 4<i>'''</i>-hydroxyphenethyl(4<i>'</i>-hydroxy-3<i>'</i>-methoxyphenyl)propionate (<b>2</b>) and 4<i>''</i>-hydroxyphenethyl ethyl succinate (<b>3</b>), together with 5 known compounds, ferulic acid (<b>4</b>), L-phenyllactic acid (<b>5</b>), hydroxy<em>tyrosol</em> (<b>6</b>), dihydroferulic acid (<b>7</b>), cyclo(L-Pro-D-Tyr) (<b>8</b>). Their structures were elucidated using spectroscopic analysis and by comparison with the literature data. All compounds displayed antioxidant effect in the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical. Among them, the new compound <b>2</b> exhibited obvious antioxidant effect, and new compounds <b>1</b> and <b>3</b> exhibited medium antioxidant effect.

Keywords: Huangjing wine; anti-oxidation; extraction and isolation; structural identification; tyrosol derivative.

Lupin seeds can represent a valuable source of phenolics and other antioxidant compounds. In this work, a comprehensive analysis of the phytochemical profile was performed on seeds from three Lupinus species, including one cultivar (Lupinus albus) and two wild accessions (Lupinus cossentinii and Lupinus luteus), collected from the northern region of Tunisia. Untargeted metabolomic profiling allowed to identify 249 compounds, with a great abundance of phenolics and alkaloids. In this regard, the species L. cossentinii showed the highest phenolic content, being 6.54 mg/g DW, followed by L. luteus (1.60 mg/g DW) and L. albus (1.14 mg/g DW). The in vitro antioxidant capacity measured by the ABTS assay on seed extracts ranged from 4.67 to 17.58 mg trolox equivalents (TE)/g, recording the highest values for L. albus and the lowest for L. luteus. The DPPH radical scavenging activity ranged from 0.39 to 3.50 mg TE/g. FRAP values varied between 4.11 and 5.75 mg TE/g. CUPRAC values for lupin seeds ranged from 7.20 to 8.95 mg TE/g, recording the highest for L. cossentinii. The results of phosphomolybdenum assay and metal chelation showed similarity between the three species of Lupinus. The acetylcholinesterase (AChE) inhibition activity was detected in each methanolic extract analyzed with similar results. Regarding the butyrylcholinesterase (BChE) enzyme, it was weakly inhibited by the Lupinus extracts; in particular, the highest activity values were recorded for L. albus (1.74 mg GALAE/g). Overall, our results showed that L. cossentinii was the most abundant source of polyphenols, consisting mainly in tyrosol equivalents (5.82 mg/g DW). Finally, significant correlations were outlined between the phenolic compounds and the in vitro biological activity measured, particularly when considering flavones, phenolic acids and lower-molecular-weight phenolics.

Keywords: Lupinus; UHPLC-QTOF-MS; acetylcholine esterase inhibition; bioactive compounds; foodomics; functional components; radical scavenging; untargeted profiling.

Tyrosol and hydroxytyrosol derived from virgin olive oil and olives extract, have wide applications both as functional food components and as nutraceuticals. However, they have low bioavailability due to their low absorption and high metabolism in human liver and small intestine. Acetylation of tyrosol and hydroxytyrosol can effectively improve their bioavailability and thus increase their potential use in the food and cosmeceutical industries. There is no report on the bioproductin of tyrosol acetate and hydroxytyrosol acetate so far. Thus, it is of great significance to develop microbial cell factories for achieving tyrosol acetate or hydroxytyrosol acetate biosynthesis. In this study, a de novo biosynthetic pathway for the production of tyrosol acetate and hydroxytyrosol acetate was constructed in Escherichia coli. First, an engineered E. coli that allows production of tyrosol from simple carbon sources was established. Four aldehyde reductases were compared, and it was found that yeaE is the best aldehyde reductase for tyrosol accumulation. Subsequently, the pathway was extended for tyrosol acetate production by further overexpression of alcohol acetyltransferase ATF1 for the conversion of tyrosol to tyrosol acetate. Finally, the pathway was further extended for hydroxytyrosol acetate production by overexpression of 4-hydroxyphenylacetate 3-hydroxylase HpaBC.

Keywords: Escherichia coli; Hydroxytyrosol acetate; Metabolic engineering; Tyrosol acetate.

Hydroxytyrosol is an important fine chemical and is widely used in food and medicine as a natural antioxidant. Production of hydroxytyrosol through synthetic biology is of important significance. Here we cloned and functionally characterized a hydroxylase encoding gene HpaBC from Escherichia coli BL21, and both subunits of this enzyme can be successfully expressed to convert the tyrosol into hydroxytyrosol. A HpaBC gene integration expression cassette under the tac promoter was constructed, and integrated into the genome of a tyrosol hyper-producing E. coli YMG5A*R using CRISPR-Cas9 technology. Meanwhile, the pathway for production of acetic acid was deleted, resulting in a recombinant strain YMGRD1H1. Shake flask fermentation showed that strain YMGRD1H1 can directly use glucose to produce hydroxytyrosol, reaching a titer of 1.81 g/L, and nearly no by-products were detected. A titer of 2.95 g/L was achieved in a fed-batch fermentation conducted in a 5 L fermenter, which is the highest titer for the de novo synthesis of hydroxytyrosol from glucose reported to date. Production of hydroxytyrosol by engineered E. coli lays a foundation for further construction of hydroxytyrosol cell factories with industrial application potential, adding another example for microbial manufacturing of aromatic compounds.

Keywords: CRISPR-Cas9; Escherichia coli; hydroxylase; hydroxytyrosol; tyrosol.

Tremella fuciformis is a dimorphic fungus which can undertake the reversible transition between yeast and pseudohypha forms. G protein α subunit (Gα) carries different signals to regulate a variety of biological processes in eukaryotes, including fungal dimorphism. In this study, a novel Gα subunit encoded gene, TrGpa1, was firstly cloned from T. fuciformis. The TrGpa1 open reading frame has 1059 nucleotides, and encodes a protein which belongs to the group I of Gα_i superfamily. Furthermore, the role of TrGpa1 in the T. fuciformis dimorphism was analysed by gene overexpression and knockdown. Stable integration of the target gene into the genome was confirmed by PCR and Southern blot hybridization. Transformants with the highest and lowest TrGpa1 expression levels were selected via quantitative real-time PCR analysis and Western blot. Each transformant was compared with the wild-type strain about the morphological change under different environmental factors, including pH values, temperature, cultivation time, inoculum size, and quorum-sensing molecules (farnesol and tyrosol). Comparing with the wild-type strain, the overexpression transformant always had higher ratios of pseudohyphae, while the knockdown transformant had less proportions of pseudohyphae. Therefore, the TrGpa1 is involved in the dimorphism of T. fuciformis and plays a positive role in promoting pseudohyphal growth.

Keywords: Dimorphism; G protein α subunit; Gene function; Morphological change; Tremella fuciformis.

Objectives: The present study aimed to determine the phenolic compounds present in the water-soluble extracts of Thymus munbyanus subsp. ciliatus using high pressure liquid chromatography-time-of-flight mass spectrometry (MS). These phenolic compounds were further isolated and characterized for their antioxidant activities.

Materials and methods: The aerial parts of T. munbyanus subsp. ciliatus were air dried, powdered, and extracted using water:methanol three times. The concentrated hydromethanolic extract was further dissolved in H₂O, filtered, and successively extracted using ethyl acetate, chloroform, and n-butanol. T. munbyanus extracts were further purified using column chromatography, and the purified extracts were subjected to in vitro antioxidant assays.

Results: Two previously undescribed compounds, namely methyl 2,3,5,6-tetrahydroxybenzoate and 4-hydroxy-5-methoxy-2-oxo-2H-pyran-3-carboxylic acid, and 14 known compounds, including 3 flavonoids; namely 3',5,5',7-tetrahydroxyflavanone, luteolin, and isorhamnetin-3-O-β-glucoside; a sterol glucoside named daucosterol; and 10 phenolic compounds, namely salicylic acid, ferulic acid, pluchoic acid, ethyl caffeate, methyl caffeate, protocatechuic acid, rosmarinic acid, p-coumaric acid, tyrosol, and protocatechuic aldehyde, were isolated from ethyl acetate and n-butanol extracts. The isolated compounds were characterized using 1D-2D^-1H^-13C nuclear magnetic resonance and MS methods.

Conclusion: The compounds isolated from ethyl acetate and n-butanol extracts exhibited excellent antioxidant and 2,2-diphenyl-1-picrylhydrazyl scavenging activities. All these results highlighted the antioxidant potential of the isolated phenolic compounds and extracts, which could be further utilized for different pharmacological applications.

Keywords: Thymus munbyanus subsp. ciliatus; antioxidant activity; isolated compounds; phenolics.

Cydonia oblonga Mill., normally known as the quince fruit, has been widely used in agro-food industries mainly to produce jams and jellies. However, other parts of the plants are still underutilized and not completely assessed for their nutraceutical profile. Therefore, in this work, the polyphenolic profile of C. oblonga was investigated using an untargeted metabolomics approach based on high-resolution mass spectrometry. Several compounds were identified in the different parts of the plants, including flavonoids (i.e., anthocyanins, flavones, flavan-3-ols, and flavonols), phenolic acids (both hydroxycinnamics and hydroxybenzoics), low-molecular-weight phenolics (tyrosol equivalents), lignans, and stilbenes. Overall, C. oblonga leaves showed the highest in vitro antioxidant potential, as revealed by 2,2-difenil-1-picrylhydrazyl (DPPH), 2,2'-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP), and cupric ion reducing antioxidant capacity (CUPRAC) assays, being 189.5, 285.6, 158.9, and 348.8 mg Trolox Equivalent/g, respectively. The enzymes acetyl- and butyryl-cholinesterases were both inhibited by the different plant parts of C. oblonga, with stems showing the higher inhibitory potential. Interestingly, the fruit extracts were the only parts inhibiting the α-glucosidase, with a value of 1.36 mmol acarbose equivalents (ACAE)/g. On the other hand, strong tyrosinase inhibition was found for stems and leaves, being 72.11 and 68.32 mg Kojic acid Equivalent/g, respectively. Finally, a high number of significant (0.05 < p < 0.01) correlations were outlined between phenolics (mainly anthocyanins, flava-3-ols, and tyrosol equivalents) and the different biological assays. Taken together, our findings suggest a potential exploitation of C. oblonga leaves and stems for the food, pharmaceutical, and cosmetic industries.

Keywords: Cydonia oblonga; antioxidants; enzyme inhibitions; foodomics; nutraceuticals; polyphenols.

The INFOGEST standardized method was applied to assess the potential bioaccessibility and bioaccessibility of the phenolic compounds from a Galician extra-virgin olive oil (EVOO). The in vitro digestion model involves three steps and generates two fractions after each one: an aqueous fraction (namely, water phase (Wp)) and an oily fraction (namely, oily phase (Op)). The results showed that secoiridoids were the most abundant family in the Galician EVOO polar fraction, representing 98% of the total phenolic compounds. After oral digestion, phenolic acids and simple phenols were mainly detected in Wp, while lignans and flavonoids were mostly found in Op. After gastric digestion, extensive hydrolysis of secoiridoids was observed to generate free tyrosol, hydroxytyrosol, and hydroxytyrosol acetate. The instability of secoiridoids after intestinal digestion was again responsible for the release of simple phenols, which were mainly recovered in Wp together with flavonoids. In contrast, lignans were stable to duodenal conditions and remained in Op.

Keywords: antioxidant capacity; bioaccessibility; extra-virgin olive oil; in vitro digestion; phenolic compounds; α-glucosidase inhibition.

Hydroxytyrosol (HT) is the component primarily responsible for the neuroprotective effect of extra virgin olive oil (EVOO). However, it is less effective on its own than the demonstrated neuroprotective effect of EVOO, and for this reason, it can be postulated that there is an interaction between several of the polyphenols of EVOO. The objective of the study was to assess the possible interaction of four EVOO polyphenols (HT, tyrosol, dihydroxyphenylglycol, and oleocanthal) in an experimental model of hypoxia-reoxygenation in rat brain slices. The lactate dehydrogenase (LDH) efflux, lipid peroxidation, and peroxynitrite production were determined as measures of cell death, oxidative stress, and nitrosative stress, respectively. First, the polyphenols were incubated with the brain slices in the same proportions that exist in EVOO, comparing their effects with those of HT. In all cases, the cytoprotective and antioxidant effects of the combination were greater than those of HT alone. Second, we calculated the concentration-effect curves for HT in the absence or presence of each polyphenol. Tyrosol did not significantly modify any of the variables inhibited by HT. Dihydroxyphenylglycol only increased the cytoprotective effect of HT at 10 µM, while it increased its antioxidant effect at 50 and 100 µM and its inhibitory effect on peroxynitrite formation at all the concentrations tested. Oleocanthal increased the cytoprotective and antioxidant effects of HT but did not modify its inhibitory effect on nitrosative stress. The results of this study show that the EVOO polyphenols DHPG and OLC increase the cytoprotective effect of HT in an experimental model of hypoxia-reoxygenation in rat brain slices, mainly due to a possibly synergistic effect on HT's antioxidant action. These results could explain the greater neuroprotective effect of EVOO than of the polyphenols alone.

Keywords: extra virgin olive oil; hydroxytyrosol; neuroprotection; polyphenols.

A considerable amount of literature has been published claiming the cardiovascular benefits of moderate (red) wine drinking, which has been considered a distinguishing trait of the Mediterranean diet. Indeed, red wine contains relevant amounts of polyphenols, for which evidence of their biological activity and positive health effects are abundant; however, it is also well-known that alcohol, even at a low level of intake, may have severe consequences for health. Among others, it is directly related to a number of non-communicable diseases, like liver cirrhosis or diverse types of cancer. The IARC classifies alcohol as a Group 1 carcinogen, causally associated with the development of cancers of the upper digestive tract and liver, and, with sufficient evidence, can be positively associated with colorectum and female breast cancer. In these circumstances, it is tricky, if not irresponsible, to spread any message on the benefits of moderate wine drinking, about which no actual consensus exists. It should be further considered that other hallmarks of the Mediterranean diet are the richness in virgin olive oil, fruits, grains, and vegetables, which are also good sources of polyphenols and other phytochemicals, and lack the risks of wine. All of these aspects are reviewed in this article.

Keywords: alcohol; olive oil; phytochemicals; resveratrol; tyrosol.

Olive mill wastewater (OMW) contains valuable and interesting bioactive compounds, among which is hydroxytyrosol, which is characterized by a remarkable antioxidant activity. Due to the health claims related to olive polyphenols, the aim of this study was to obtain an extract from OMW with an increased level of hydroxytyrosol by means of microbial enzymatic activity. For this purpose, four commercial adsorbent resins were selected and tested. The beta-glucosidase and esterase activity of strains of Wickerhamomyces anomalus, Lactiplantibacillus plantarum, and Saccharomyces cerevisiae were also investigated and compared to those of a commercial enzyme and an Aspergillus niger strain. The W. anomalus strain showed the best enzymatic performances. The SP207 resin showed the best efficiency in selective recovery of hydroxytyrosol, tyrosol, oleuropein, and total phenols. The bioconversion test of the OMW extract was assessed by using both culture broths and pellets of the tested strains. The results demonstrated that the pellets of W. anomalus and L. plantarum were the most effective in hydroxytyrosol increasing in phenolic extract. The interesting results suggest the possibility to study new formulations of OMW phenolic extracts with multifunctional microorganisms.

Keywords: Lactiplantibacillus plantarum; Wickerhamomyces anomalus; adsorbent resins; beta-glucosidase; esterase; hydroxytyrosol; oleuropein; tyrosol.

Dietary polyphenols have been the focus of major interest for their potential benefits on human health. Several preclinical studies have been conducted to provide a rationale for their potential use as therapeutic agents in preventing or ameliorating cognitive decline. However, results from human studies are scarce and poorly documented. The aim of this review was to discuss the potential mechanisms involved in age-related cognitive decline or early stage cognitive impairment and current evidence from clinical human studies conducted on polyphenols and the aforementioned outcomes. The evidence published so far is encouraging but contrasting findings are to be taken into account. Most studies on anthocyanins showed a consistent positive effect on various cognitive aspects related to aging or early stages of cognitive impairment. Studies on cocoa flavanols, resveratrol, and isoflavones provided substantial contrasting results and further research is needed to clarify the therapeutic potential of these compounds. Results from other studies on quercetin, green tea flavanols, hydroxycinnamic acids (such as chlorogenic acid), curcumin, and olive oil tyrosol and derivatives are rather promising but still too few to provide any real conclusions. Future translational studies are needed to address issues related to dosage, optimal formulations to improve bioavailability, as well as better control for the overall diet, and correct target population.

Keywords: Anthocyanins; Catechins; Cognitive decline; Cognitive impairment; Nutraceuticals; Polyphenols; Resveratrol; chlorogenic acid; isoflavones; quercetin.