A silica-chitosan support was employed for laccase immobilization. The hybrid support was obtained using calcium ion as linking agent that coordinates silanol and hydroxyl groups of chitosan. The insoluble biocatalyst was then packed in a column and used in a flow system for phenol removal. The immobilized enzyme reactor (IMER) showed a good storage stability (70% of activity in 70 days) and good reusability (90-50% of catalytic activity at the 4th reuse in function of chitosan type). The best performance for the phenol removal was obtained with a low molecular weight chitosan from crab shells at pH 5 and with a flow rate of 0.7 mL/min. The apparent Michaelis-Menten (V_max^app, K_m^app) and the inherent (V_max^inh, K_m^inh) constants were also determined to evaluate the influence of the phenol structure on the performance of the system. The enzymatic oxidation of a phenol mixture (4-methylcatechol, catechol, caffeic acid, syringic acid, vanillic acid, p-coumaric acid, and tyrosol) was followed for 21 h in a continuous mode by HPLC. The phenol mixture removal of 90% was also confirmed by Folin-Ciocalteu assay.

Keywords: continuous mode; immobilized laccase; silica-chitosan support.

Two series of 3,5-disubstituted isoxazoles (6a-e) and 1,4-disubstituted triazoles (8a-e) derivatives have been synthesized from tyrosol (1), a natural phenolic compound, detected in several natural sources such as olive oil, and well-known by its wide spectrum of biological activities. Copper-catalyzed microwave-assisted 1,3-dipolar cycloaddition reactions between tyrosol-alkyne derivative 2 and two series of aryl nitrile oxides (5a-e) and azides (7a-e) regiospecifically afforded 3,5-disubstituted isoxazoles (6a-e) and 1,4-triazole derivatives (8a-e), respectively in quantitative yields. Synthesized compounds were purified and characterized by spectroscopic means including 1D and 2D NMR techniques and HRMS analysis. The newly prepared hybrid molecules have been evaluated for their anticancer and hemolytic activities. Results showed that most derivatives displayed significant antiproliferative activity against human glioblastoma cancer cells (U87) in a dose-dependent manner. Compounds 6d (IC₅₀ = 15.2 ± 1.0 μg/mL) and 8e (IC₅₀ = 21.0 ± 0.9 μg/mL) exhibited more potent anticancer activity. Moreover, most derivatives displayed low hemolytic activity, even at higher concentrations which suggested that these classes of compounds are suitable candidates for further in vivo investigations. The obtained results allow us to consider the newly synthesized isoxazole- and triazole-linked tyrosol derivatives as promising scaffolds for the development of effective anticancer agents.

Keywords: Antiproliferative activity; Isoxazoles; Microwave-assisted synthesis; Triazoles; Tyrosol.

Extra virgin olive oil (EVOO) is a known source of antioxidants, such as phenolic compounds, useful in the prevention of non-infectious diseases (atherosclerosis, diabetes, cancer, and other diseases). In the present study, EVOO obtained using an innovative ultrasounds-based technology was found richer in total polyphenols, hydroxytyrosol and tyrosol, than EVOO obtained using a conventional mechanical technology. The urinary excretion in humans of hydroxytyrosol and tyrosol, after the administration of ultrasounds and mechanical EVOOs, respectively, was assessed and compared. The analytes were determined in urine samples, collected for 24 h, of six healthy people (3 men and 3 women, age 22-70 years and body mass index <30 kg/m²) who ingested 20 g of oil for six consecutive days. A commercial refined olive oil was also used in the study to determine the baseline excretion levels of the two metabolites. High correlation coefficients (≥0.9311) were found between the amounts of the analytes ingested daily with EVOOs and those determined in the 24-h urines. The results clearly indicated that the EVOO obtained with the ultrasound process was characterized by the highest concentration of biophenols which were consequently available in greater quantities after ingestion, indicating that it represents a high-quality product containing high levels of beneficial compounds such as biophenols readily assimilable by the human body.

Keywords: Extra virgin olive oil; Hydroxytyrosol; Polyphenols; Tyrosol; Ultrasound olive oil extraction; Urine.

Table olives, a product of olive tree (Olea europaea L.), is an important fermented product of the Mediterranean Diet. Agronomical factors, particularly the cultivar, the ripening stage and the processing method employed are the main factors influencing the nutritional and non-nutritional composition of table olives and their organoleptic properties. The important nutritional value of this product is due to its richness in monounsaturated fat (MUFA), mainly oleic acid, fibre and vitamin E together with the presence of several phytochemicals. Among these, hydroxytyrosol (HT) is the major phenolic compound present in all types of table olives. There is a scarcity of in vitro, in vivo and human studies of table olives. This review focused comprehensively on the nutrients and bioactive compound content as well as the health benefits assigned to table olives. The possible health benefits associated with their consumption are thought to be primarily related to effects of MUFA on cardiovascular health, the antioxidant (AO) capacity of vitamin E and its role in protecting the body from oxidative damage and the anti-inflammatory and AO activities of HT. The influence of multiple factors on composition of the end product and the potential innovation in the production of table olives through the reduction of its final salt content was also discussed.

Keywords: ALS, amyotrophic lateral sclerosis; AO, antioxidant; Alpha-tocopherol; BP, blood pressure; CVD, cardiovascular disease; DM-II, Diabetes Mellitus 2; EFSA, European Food Safety Authority; FM, fat mass; GSH, glutathione; HDL-c, high-density lipoprotein cholesterol; HT, hydroxytyrosol; LDL-c, low-density lipoprotein cholesterol; MD, Mediterranean Diet; MUFA, monounsaturated fat; Mediterranean Diet; Monounsaturated fat; NO, nitric oxide; NaCl, sodium chloride; NaOH, sodium hydroxide; Nrf2, nuclear factor erythroid 2-related factor 2; OL, oleuropein; OO, olive oil; PKC, protein kinase C; PUFA, polyunsaturated fat; Phenolic compounds; RDA, Recommended Dietary Allowance; ROS, reactive oxygen species; TC, total cholesterol; TG, triacylglycerol; TG, triglyceride; Table olives; Ty, tyrosol; WHO, World Health Organization; cv, cultivar; e.p, edible portion; α-TOH, alpha-tocopherol.

The necessity for developing novel cytostatic agents with improved activities and reduced side-effects to tackle cancer prompted us to investigate mitochondria-targeted compounds, an approach that is gaining attention for the selective transportation of cytotoxic agents. We envisioned the possibility of conjugating a phenethyl alcohol motif, decorated with a series of phenol-based substituents on the aryl moiety, with a triphenyl phosphonium scaffold (a mitochondria-directed vector), through a hydrocarbon chain of different lengths. Thus, such compounds that incorporate the phenethyl skeleton can be considered as masked phenolic compounds derived from relevant natural counterparts found in olive tree (e.g. tyrosol, hydroxytyrosol). Title compounds exhibited very strong in vitro antiproliferative activities against the panel of six human tumor cell lines tested, with GI₅₀ values ranging from the nanomolar (0.026 ± 0.010 μM for 36) to the submicromolar range in most of the cases; this represents an improvement of up to 350-fold compared to classical chemotherapeutic agents, like 5-fluorouracil or cisplatin. Interestingly, decrease in the linker length led to an increase of GI₅₀ values against non-tumor cells, thus allowing a remarkable improvement of selectivity (SI up to 269). The very promising antiproliferative activities prompted us to further investigate their behaviour against multidrug resistant cell lines (MDR). The results indicated a reduced sensitivity of the multidrug resistant cells to compounds, probably due to P-gp-mediated efflux of these antiproliferative agents. Interestingly, activities were completely restored to the same levels by co-administration of tariquidar, a well-known inhibitor of P-gp. Flow cytometry analysis on sensitive cell lines revealed a decrease in the percentage of cells in G1 phase accompanied by increase in S and G2/M phases. In addition, a significant increase in subG1 area, was observed. These results are compatible with the necrotic and apoptotic cell death detected in the Annexin V assay, and with the depolarization of the mitochondria membrane. Thus, the new mitochondriotropic agents reported herein can be considered as promising antiproliferative agents, endowed with remarkable potency and selectivity, including MDR cells, upon co-administration with a pump-efflux inhibitor.

Keywords: Antiproliferative agents; Chemosensitizer; Mitocans; Mitochondriotropics; Multidrug resistant cells; Phosphonium salts.

The present study delineates the effects of incorporation of 1% diallyl sulfide (DAS) into extra virgin olive oil (EVOO) on the physico-chemical characteristics, in vitro antioxidant, and in vivo hepatoprotective properties in CCl₄-induced acute liver injury in mice. Results showed that the DAS-rich EVOO exhibited good oxidative stability over one-month storage and preserved its original quality-related parameters including major components (oleic acid, linoleic acid, and palmitic acid), and minor components (tocopherols, chlorophylls and carotenoids, tyrosol, hydroxytyrosol, elenolic acid, oleuropein and its aglycone, pinoresinol, vanilic acid, cinnamic acid, ferulic acid, luteolin, apigenin, and sterols). Compared with EVOO or DAS, the DAS-rich EVOO displayed the highest DPPH and ABTS-radical scavenging activities and showed the strongest cellular antioxidant activity (CAA). In connection with its free radical scavenging activity and CAA, DAS-rich EVOO significantly normalized the serum ALT and AST levels and prevented the increase in interleukin-6 in CCl₄-intoxicated mice. The manifest anti-inflammatory and hepatoprotective effects of DAS-rich EVOO were further supported by liver histopathological examinations. Overall, the EVOO enrichment with DAS could open up opportunities for the development of novel functional food with improved antioxidant and hepatoprotective properties.

Keywords: carbon tetrachloride; cellular antioxidant activity; diallyl sulfide‐rich extra virgin olive oil; functional foods; inflammation; liver injury.

Tyrosol (T) and hydroxytyrosol (HOT) and their glycosides are promising candidates for applications in functional food products or in complementary therapy. A series of phenylethanoid glycofuranosides (PEGFs) were synthesized to compare some of their biochemical and biological activities with T and HOT. The optimization of glycosylation promoted by environmentally benign basic zinc carbonate was performed to prepare HOT α-L-arabino-, β-D-apio-, and β-D-ribofuranosides. T and HOT β-D-fructofuranosides, prepared by enzymatic transfructosylation of T and HOT, were also included in the comparative study. The antioxidant capacity and DNA-protective potential of T, HOT, and PEGFs on plasmid DNA were determined using cell-free assays. The DNA-damaging potential of the studied compounds for human hepatoma HepG2 cells and their DNA-protective potential on HepG2 cells against hydrogen peroxide were evaluated using the comet assay. Experiments revealed a spectrum of different activities of the studied compounds. HOT and HOT β-D-fructofuranoside appear to be the best-performing scavengers and protectants of plasmid DNA and HepG2 cells. T and T β-D-fructofuranoside display almost zero or low scavenging/antioxidant activity and protective effects on plasmid DNA or HepG2 cells. The results imply that especially HOT β-D-fructofuranoside and β-D-apiofuranoside could be considered as prospective molecules for the subsequent design of supplements with potential in food and health protection.

Keywords: DNA damage; DNA-protective effect; antioxidant activity; basic zinc carbonate; cytotoxicity; hydroxytyrosol; phenylethanoid glycofuranosides; tyrosol.

Nitric oxide (NO) is an important signaling molecule involved in many pathophysiological processes. NO mediates vasodilation and blood flow in the arteries, and its action contributes to maintaining vascular homeostasis by inhibiting vascular smooth muscle contraction and growth, platelet aggregation, and leukocyte adhesion to the endothelium. Dietary antioxidants and their metabolites have been found to be directly and/or indirectly involved in the modulation of the intracellular signals that lead to the production of NO. The purpose of this study was to investigate the contribution of conjugated metabolites of hydroxytyrosol (HT) and tyrosol (TYR) to the release of NO at the vascular level, and the related mechanism of action, in comparison to their parental forms. Experiments were performed in human aortic endothelial cells (HAEC) to evaluate the superoxide production, the release of NO and production of cyclic guanosine monophosphate (cGMP), the activation of serine/threonine-protein kinase 1 (Akt1), and the activation state of endothelial nitric oxide synthase (eNOS). It was observed that the tested phenolic compounds enhanced NO and cGMP concentration, inhibiting its depletion caused by superoxide overproduction. Moreover, some of them enhanced the activation of Akt (TYR, HT metabolites) and eNOS (HT, HVA, TYR-S, HT-3S). Overall, the obtained data showed that these compounds promote NO production and availability, suggesting that HT and TYR conjugated metabolites may contribute to the effects of parental extra virgin olive oil (EVOO) phenolics in the prevention of cardiovascular diseases.

Keywords: conjugate metabolites; glucuronide; hydroxytyrosol; nitric oxide; olive oil; phenolic acids; sulfate; superoxide; tyrosol.

We investigated the antihyperlipidemic effects of tyrosol in streptozotocin (STZ)-induced diabetic rats. Rats were injected intraperitoneally with STZ (40 mg/kg), and these established experimental rats were treated with tyrosol (20 mg/kg) and glibenclamide (600µg/kg) for 45 days. The observed results revealed that tyrosol treatment significantly reduced plasma glucose, plasma, and liver total cholesterol, triglycerides, free fatty acids, phospholipids, plasma low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, atherogenic index, and significantly increased plasma insulin and high-density lipoprotein cholesterol in STZ-induced diabetic rats. The activity of3-hydroxy 3-methylglutaryl coenzyme A reductase significantly reduced in the liver, whereas the activities of lipoprotein lipase and lecithin cholesterol acyltransferase were significantly increased in the plasma of tyrosol treated STZ-induced diabetic rats. Histological examination showed that tyrosol treatment remarkably reduced lipid accumulation in the liver of STZ-induced diabetic rats. The present study revealed that tyrosol exhibits potent antihyperlipidemic effects in STZ-induced diabetic rats.

Keywords: Diabetes Mellitus; Insulin; Lipids; Streptozotocin; Tyrosol.

Rhodiola rosea L. rhizome has been used as a traditional medicine to treat fatigue, depression, and cognitive dysfunction. We aimed to authenticate R. rosea L. rhizome using the DNA barcoding technique and to quantify its main compounds, total phenolics, total flavonoids, and antioxidant capacity, and then to investigate their neuroprotective effects. The sequences of internal transcribed spacer and trnH-psbA of R. rosea L. rhizomes showed a 99% identity with those of NCBI GenBank database according to BLAST searches. Analysis using reversed-phase HPLC revealed five main compounds in R. rosea L. rhizome. Rhodiola rosea L. rhizome and two bioactive compounds, salidroside and tyrosol, showed free radical scavenging activity. Rhodiola rosea L. rhizome and its identified compounds protected neuronal PC-12 cells against oxidative stress and showed moderate acetylcholinesterase inhibition. Taken together, these results suggest that R. rosea L. rhizomes with bioactives can be used as a functional ingredient with potential for neuroprotection.

Supplementary information: The online version of this article (doi:10.1007/s10068-020-00868-7) contains supplementary material, which is available to authorized users.

Keywords: Acetylcholinesterase; DNA barcoding; Oxidative stress; Salidroside; Tyrosol.

Extra virgin olive oil (EVOO) is well known for containing relevant amounts of healthy phenolic compounds. The European Food Safety Authority (EFSA) allowed a health claim for labelling olive oils containing a minimum amount of hydroxytyrosol (OHTyr) and its derivatives, including tyrosol (Tyr). Therefore, harmonized and standardized analytical protocols are required in support of an effective application of the health claim. Acid hydrolysis performed after extraction and before chromatographic analysis has been shown to be a feasible approach. Nevertheless, other fast, green, and easy methods could be useful for on-site screening and monitoring applications. In the present research, a natural deep eutectic solvent (NADES) composed of lactic acid and glucose was used to perform a liquid/liquid extraction on EVOO samples, followed by UV-spectrophotometric analysis. The spectral features of the extracts were related with the content of total OHTyr and Tyr, determined by the acid hydrolysis method. The second derivative of spectra allowed focusing on three single wavelengths (i.e., 299 nm, 290 nm, and 282 nm) significantly related with total OHTyr, total Tyr, and their sum, respectively. In particular, the sum of OHTyr and Tyr could be determined with a root mean square error of prediction of 29.5 mg kg^-1, while the limits of quantitation and detection were respectively 11.8 and 4.9 mg kg^-1. The proposed method, therefore, represents an easy screening tool, with the use of a green, food-derived solvent, and could be considered as an attempt to pave the way for food grade analytical chemistry.

Keywords: acidic hydrolysis; derivative UV spectroscopy; green chemistry; health claim; phenolic compounds; screening methods.

European virgin olive oil with geographical indications are strictly regulated and are of importance for the producing member states, Spain, Italy, Greece, Portugal, France, Slovenia and Croatia. These products are consumed locally, or within the European Union but are also exported worldwide. The chapter stresses on the importance of combining origin indications with other certifications or opportunities raising from European legislation in the agri-food sector so that to tighten consumer loyalty for this category of products. Emphasis is given to the richness of virgin olive oil in bioactive compounds that are already covered by nutritional and health claims (oleic acid, vitamin E, "polyphenols") and to those compounds that can be exploited in the future toward the same direction (squalene, oleanolic and maslinic acids).

Keywords: Bioactives; Geographical indications; Nutritional and health claims; PDO/PGI; Quality schemes; Squalene; Total hydroxytyrosol and tyrosol content; Triterpenic acids; Virgin olive oil; Vitamin E.

In this study, four Brassicaceae microgreens species, namely kale, red cabbage, kohlrabi, and radish, were evaluated for their phytochemical compositions using spectrophotometric assays and untargeted metabolomics before and after in vitro gastrointestinal digestion. According to the in vitro spectrophotometric results, significant amounts of phenolics could be detected in each studied species, thus supporting the total antioxidant capacities recorded. Overall, metabolomics allowed annotating a total of 470 phytochemicals across the four Brassicaceae microgreens, either fresh or digested. Among polyphenols, flavonoids were the most represented class (180 compounds, including anthocyanins, flavones, flavonols, and other flavonoids), followed by phenolic acids (68 compounds, mainly hydroxycinnamic and hydroxybenzoic acids), non-flavonoid or phenolic acid-based structures (i.e., alkyl- and alkylmethoxy-phenols and tyrosol derivatives), and lignans. Also, 22 glucosinolates were annotated, including gluconapin glucoraphanin, glucobrassicin, and 4-hydroxyglucobrassicin. Noteworthy, significant differences could be observed in terms of bioaccessibility as a function of the phenolic class and the species considered. Overall, lignans exhibited the highest bioaccessibility values (14%), followed by tyrosol derivatives and flavonoids (on average, 9% and 8%, respectively). However, differences could be evidenced as a function of the species, with red cabbage having comparatively lower bioaccessibility values irrespective of the chemical class of bioactive considered. Similarly, bioaccessibility of glucosinolates significantly differed across species, ranging from 2% in kale to 43% in kohlrabi microgreens.

Keywords: Bioaccessibility; Enzyme activity; Food metabolomics; In vitro antioxidant capacity; Phenolic compounds.

Two extra virgin olive oil (EVOO) phenolic extracts (BUO and OMN) modulate DPP-IV activity. The in vitro DPP-IV activity assay was performed at the concentrations of 1, 10, 100, 500, and 1000 μg/mL, showing a dose-dependent inhibition by 6.8 ± 1.9, 17.4 ± 6.1, 37.9 ± 2.4, 57.8 ± 2.9, and 81 ± 1.4% for BUO and by 5.4 ± 1.7, 8.9 ± 0.4, 28.4 ± 7.2, 52 ± 1.3, and 77.5 ± 3.5% for OMN. Moreover, both BUO and OMN reduced the DPP-IV activity expressed by Caco-2 cells by 2.9 ± 0.7, 44.4 ± 0.7, 61.2 ± 1.8, and 85 ± 4.2% and by 3 ± 1.9, 35 ± 9.4, 60 ± 7.2, and 82 ± 2.8%, respectively, at the same doses. The concentration of the most abundant and representative secoiridoids within both extracts was analyzed by nuclear magnetic resonance (¹H-NMR). Oleuropein, oleacein, oleocanthal, hydroxytyrosol, and tyrosol, tested alone, reduced the DPP-IV activity, with IC₅₀ of 472.3 ± 21.7, 187 ± 11.4, 354.5 ± 12.7, 741.6 ± 35.7, and 1112 ± 55.6 µM, respectively. Finally, in silico molecular docking simulations permitted the study of the binding mode of these compounds.

Keywords: Caco-2 cells; DPP-IV; EVOO; phenolic extracts; secoiridoids.

Glycemic control is a mainstay of type 2 diabetes mellitus (T2DM) clinical management. Despite the continuous improvement in knowledge and progress in terms of treatment, the achievement of the physiologic metabolic profile is still an ongoing challenge in diabetic patients. Pancreatic β-cell line INS-1 832/13 was used to assess the insulin secretagogue activity of hydroxytyrosyl oleate (HtyOle) and tyrosyl oleate (TyOle), two naturally occurring lipophenols deriving from the conjugation of oleic acid (OA) and hydroxytyrosol (Hty) or tyrosol (Ty), respectively. The insulin secretion was determined under a glucose-induced insulin secretion (GSIS) condition by the ELISA method. The potential involvement of G-protein-coupled receptor 40 (GPR40), also known as free fatty acid receptor 1 (FFAR1), was investigated by both molecular docking and functional pharmacological approaches. Herein, we demonstrated that HtyOle and TyOle exerted a facilitatory activity on insulin secretion under the GSIS condition. Moreover, we provided evidence that both lipophenols are natural modulators of FFAR1 receptor. From our results, the anti-diabetes properties associated with olive oil consumption can be partly explained by the HtyOle and TyOle effects.

Keywords: FFAR1; GSIS; hydroxytyrosol; insulin secretion; lipophenols; polyphenol; tyrosol.

The extraction of phenolic compounds from olive mill wastes is important, not only to avoid environmental damages, but also because of the intrinsic value of those biophenols, well-known for their high antioxidant potential and health benefits. This study focuses on tyrosol (Tyr) and hydroxytyrosol (HT), two of the main phenolic compounds found in olive mill wastes. A new, simple, and eco-friendly extraction process for the removal of phenolic compounds from aqueous solutions using native β-cyclodextrin (β-CD) in the solid state has been developed. Several β-CD/biophenol molar ratios and biophenol concentrations were investigated, in order to maintain β-CD mostly in the solid state while optimizing the extraction yield and the loading capacity of the sorbent. The extraction efficiencies of Tyr and HT were up to 61%, with a total solid recovery higher than 90% using an initial concentration of 100 mM biophenol and 10 molar equivalents of β-CD. The photochemical stability of the complexes thus obtained was estimated from ∆E*ab curve vs. illumination time. The results obtained showed that the phenols encapsulated into solid β-CD are protected against photodegradation. The powder obtained could be directly developed as a safe-grade food supplement. This simple eco-friendly process could be used for extracting valuable biophenols from olive mill wastewater.

Keywords: extraction; hydroxytyrosol; tyrosol; β-cyclodextrin.

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.