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The study of drug synergy plays a prominent role in the search for drug combinations with beneficial interactions. Firstly, in this process, the drug-effect response of individual parts and the mixture needs to be derived. This function is usually well described by Hill (or other logistic or sigmoid) curve. Due to its boundedness, it allows the measured data to be normalized. The normalized data can then be processed by interaction analysis using the Loewe, Bliss, or other models to evaluate possible synergy or antagonism of two or more drugs. However, sometimes, the drug-effect responses observed in pharmaceutical research do not appear to be bounded. Theoretically, the drug-effect curve cannot grow to infinity, but it may be impossible to determine its upper bound within the observed region. In this case, standard models cannot be used, since they assume that data are normalized. The approach of this article bypasses the need to normalize the data, allowing its broader application and usefulness in finding potential synergies in pharmaceutical research.

Resveratrol and caffeic acid belong to plant polyphenols and are known for their antioxidant effects. The aim of our research was to study their impact on Maillard reaction. This one occurs when the reducing saccharides react with amino groups of biomolecules including proteins, alter their protein conformation and transform to the variety of advanced glycation end products (AGEs). AGEs exhibit browning and generate fluorescence. There exist expectations that this oxidative protein glycosylation could be prevented by antioxidants. In this study, we incubated bovine serum albumin (BSA) with glucose for 7 days at 37°C and measured characteristic fluorescence and UV absorbance of the formed AGEs. Surprisingly, resveratrol and caffeic acid enhanced transformation of BSA to glycation products, which was confirmed either when cupric Cu(II) or ferric Fe(III) ions in nanomolar concentration were added to the system as pro-oxidant agent.

Peppermint belongs to one of most popular medicinal plants in pharmacy as well as in the food industry. Aim: For the conventional usage, the aerial part, especially leaves, is used. This investigation was aimed at the determination of phenolic compound in peppermint rhizomes infusion and the comparison with the phenolics in leaves infusions. Methods: For the separation and identification of the phenolic compounds, the Sykam HPLC-DAD connected with Microsaic 4500MiD®, a single quadrupole mass spectrometer, was used. Results: Three compounds in rhizomes and eight compounds in leaves were identified and quantified. In rhizomes, rosmarinic acid was determined as the main secondary metabolite, but its content was three times lower than that in leaves. Infusion of peppermint leaves was richer in flavonoids content with eriocitrin as a major phenolic compound. Conclusion: Rhizomes of peppermint may also be used as a potential source of rosmarinic acid and caffeic acid derivatives.

Exogenous insulin, used as a therapeutic agent for diabetes, forms insoluble deposits containing amyloid fibrillar structures near the administration site. We have analyzed the in vitro anti-amyloid activity of four green tea constituents: (-)-epigallocatechin gallate (EGCG), (-)-epicatechin (EC), gallic acid (GA), caffeine (CF), and their equimolar mixtures. Regarding individually tested compounds, only EGCG inhibited the fibrillization process. The individual EC, GA, and CF molecules were ineffective. The presence of EGCG in equimolar combinations with GA, EC, or CF was required for the inhibitory activity of most mixtures. Molecular docking revealed that EGCG interacts with an essential amyloidogenic region of insulin chain B. Individually inactive GA had a potentiating effect on the activity of EGCG. In contrast, EC and CF had a negative impact on the activity of the mixtures. We have observed diverse morphology and the amount of insulin amyloid aggregates formed in the presence of studied compounds. The distinct types of amyloid aggregates created in vitro in the presence of EGCG and other green tea constituents were characterized. Results indicate that the biological activity of individual molecules is not directly applicable to the pooled samples effects prediction.

The berries of Vaccinium myrtillus L. are usually collected in the wild for the purpose of being a food source. They are naturally high in phenolic compounds, which possess antioxidative properties, so the berries are therefore often labeled as &ldquo, functional foods&rdquo, This study evaluated seven samples of bilberry fruits from different locations in Slovakia for the content of the main phenolic compounds (anthocyanins, flavonoids and tannins) using European Pharmacopoeia 9 spectrophotometric methods. A thorough analysis of environmental factors showed that several phenolic constituents are closely corresponding with their respective environments, as well as with each other. The environmental factors with statistically significant correlations in this study are altitude, habitat type, sunlight exposure, and soil carbon content. Our findings suggest that the berries collected at sunny sites with no topsoil damage contain more phenolic compounds. The lowest amounts of phenolic compounds were found in samples from dense forests or with visible soil erosion and windthrow damage. The negative effect of windthrow damage on the levels of secondary metabolites in bilberry fruits has been described for the first time. This study observed no relationship between the amount of phenolic compounds and soil pH, soil nitrogen levels, or slope exposition.

Bacterial infections of skin and wounds may seriously decrease the quality of life and even cause death in some patients. One of the largest concerns in their treatment is the growing antimicrobial resistance of bacterial infectious agents and the spread of resistant strains not only in the hospitals but also in the community. This trend encourages researchers to seek for new effective and safe therapeutical agents. The pharmaceutical industry, focusing mainly on libraries of synthetic compounds as a drug discovery source, is often failing in the battle with bacteria. In contrast, many of the natural compounds, and/or the whole and complex plants extracts, are effective in this field, inactivating the resistant bacterial strains or decreasing their virulence. Natural products act comprehensively; many of them have not only antibacterial, but also anti-inflammatory effects and may support tissue regeneration and wound healing. The European legislative is in the field of natural products medicinal use formed by European Medicines Agency (EMA), based on the scientific work of its Committee on Herbal Medicinal Products (HMPC). HMPC establishes EU monographs covering the therapeutic uses and safe conditions for herbal substances and preparations, mostly based on folk medicine, but including data from scientific research. In this review, the medicinal plants and their active constituents recommended by EMA for skin disorders are discussed in terms of their antibacterial effect. The source of information about these plant products in the review is represented by research articles listed in scientific databases (Science Direct, PubMed, Scopus, Web of Science, etc.) published in recent years.

The aim of this study was to analyze the binding interactions between a common antihypertensive drug (ramipril, R) and the widely distributed plant flavonoid quercetin (Q), in the presence of human serum albumin (HSA). From the observed fluorescence spectra of the (HSA + R) system we can assume that ramipril is also one of the Site 3 ligands&mdash, similar to fusidic acid&mdash, the binding of which has been proven by RTG crystallography. Our claim is supported by near-UV CD spectroscopy, microscale themophoresis and molecular modeling. The presence of R slightly inhibited the subsequent binding of Q to HSA and, on the contrary, the pre-incubation of HSA with Q caused a stronger binding of R, most likely due to allosteric interactions. At high concentrations, R is also able to displace Q from its binding site. The dissociation constant KD for the binding of R is more than hundredfold larger than for Q, which means that R is a very weak binder to HSA. The knowledge of qualitative and quantitative parameters of R, as well as the methods used in this study, are important for future research into HSA binding. This study shows the importance of implementing other methods for KD determination. Microscale thermophoresis has proved to be a novel, practical and accurate method for KD determination on HSA, especially in cases when fluorescence spectroscopy is unable to produce usable results.

Aldose reductase inhibitors are considered to be potential therapeutic agents for chronic diabetic complications. Diabetes mellitus can be accompanied by elevated blood levels of free fatty acids, which can cause lipotoxicity. Herbal extracts and their constituents are promising agents which have the potential for alleviating these complications. Our study was focused on the influence on these effects by flower infusions from Cornus mas L. and Cornus kousa F.Buerger ex Hance. Initially, phenolic compounds were quantified in the dried flowers. Next, we studied the ability of flower infusions from both plants to inhibit aldose reductase in vitro, the protective role in the cell model of lipotoxicity, and the cytotoxic action on fibroblast cell line NIH-3T3 by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2 H-tetrazolium bromide assay. Both species are rich in phenolics; C. kousa flowers contain slightly higher amounts of phenolic acids (20.8%) and flavonoids (56.1%) than C. mas (20.2%) and (47.4%), respectively. Both extracts showed effective inhibition, expressed as half-maximal inhibitory concentration (IC50) (the concentration of inhibitor required to exhibit 50% inhibition of the enzyme reaction), of aldose reductase in non-toxic low concentrations (IC50= 3.06 μg/mL for C. mas and IC50= 2.49 μg/mL for C. kousa, respectively). In contrast, these concentrations of both extracts had almost no effects in the lipotoxicity cell model. To our knowledge, this study is the first report on C. mas and C. kousa flowers’ aldose reductase inhibitory activity and influence upon lipotoxicity.

Beneficial effects of red wine polyphenols on cardiovascular health are well known. The aim of our research was an interaction study of four red wine polyphenols – resveratrol (R), quercetin (Q), kaempferol (KF) and isorhamnetin (IR) of their ability to activate endothelial NO synthase (eNOS) promoter when used alone and in equimolar mixtures. To determine their activity, we performed a luciferase reporter gene assay on EA.hy926 cells stably transfected with a luciferase reporter gene construct containing eNOS promoter. The Bradford assay was also performed to account the cytotoxicity and/or the cell number differences. The median effect equation, as an interaction analysis evaluating synergy or antagonism of the combinations was done according to mass-action law principle. Isobolographic method was performed on selected double mixtures and dose reduction index was calculated for all mixtures. All single polyphenols activated eNOS promoter. The EC50 values were in micromolar concentrations ranging from 3.44 μM (R2 = 0.96) for kaempferol to 9.89 μM for isorhamnetin (R2 = 0.94). All mixtures activated eNOS promoter, but their interactions varied from synergy (Q+R, Q+IR+KF, Q+R+KF and Q+R+IR+KF), through additive (R+IR+KF) to antagonistic interaction (R+IR, R+KF, Q+IR, Q+KF, IR+KF and R+Q+IR). In this study, we show for the first time that red wine polyphenols activated eNOS promoter when used alone and in mixtures with different type of interactions.

This overview paper describes in its introductory part bitter-tasting herbal drugs in the context of their traditional use in the loss of appetite and digestive disorders. Then, it mentions current knowledge on signals origin in bitter taste receptors leading to different, unexpected physiological processes. Newly observed biological effects of bitter tastants are correlated with modern trends of their complex study with an aim to develop new drugs primarily acting on bitter taste receptors.Key words: T2R bitter taste receptors GPCR bitters glucose homeostasis asthma ghrelin GLP-1 solitary chemosensory cells.

Protein-binding interactions are displacement reactions which have been implicated as the causative mechanisms in many drug–drug interactions. Thus, the aim of presented study was to analyse human serum albumin-binding displacement interaction between two ligands, hypoglycaemic drug gliclazide and widely distributed plant flavonoid quercetin. Fluorescence analysis was used in order to investigate the effect of substances on intrinsic fluorescence of human serum albumin (HSA) and to define binding and quenching properties of ligand–albumin complexes in binary and ternary systems, respectively. Both ligands showed the ability to bind to HSA, although to a different extent. The displacement effect of one ligand from HSA by the other one has been described on the basis of the quenching curves and binding constants comparison for the binary and ternary systems. According to the fluorescence data analysis, gliclazide presents a substance with a lower binding capacity towards HSA compared with quercetin. Results also showed that the presence of quercetin hindered the interaction between HSA and gliclazide, as the binding constant for gliclazide in the ternary system was remarkably lower compared with the binary system. This finding indicates a possibility for an increase in the non-bound fraction of gliclazide which can lead to its more significant hypoglycaemic effect. Additionally, secondary and tertiary structure conformational alterations of HSA upon binding of both ligands were investigated using synchronous fluorescence, circular dichroism and FT-IR. Experimental data were complemented with molecular docking studies. Obtained results provide beneficial information about possible interference upon simultaneous co-administration of the food/dietary supplement and drug.

Vascular smooth muscle cell (VSMC) proliferation contributes to the development of atherosclerosis. Red wine consumption due to the polyphenol content has been reported to counteract atherosclerosis progression possibly through inhibition of VSMC proliferation, among other mechanisms. In this study we investigate the antiproliferative activity of four wine polyphenols: resveratrol, quercetin, ethyl gallate, and (+)-catechin in rat aortic VSMC. All four polyphenols inhibited serum-induced VSMC proliferation when applied as a single treatment. To further address a potential synergistic action of the investigated polyphenols, the antiproliferative effect of different combinations in equimolar, as well as equipotent ratios were quantified. The IC₅₀ values of single polyphenols regarding the inhibition of VSMC proliferation ranged from 49.58 µM to 86.06 µM. However, apparent inhibitory efficacy of each compound increased by a factor of 10.4 in the quadruple equipotent mixture, as calculated from the dose-reduction index. Thus, the effective IC₅₀ values of each of the four mixture constituents ranged from 4.76 µM to 8.27 µM. The calculated combination index (CI, where CI, =, or1 indicate synergy, additivity, or antagonism, respectively) values of equimolar combinations of the polyphenols indeed indicated mainly synergy (CI ranging from 0.24 ± 0.01 to 1.51 ± 0.13). Optimized equipotent mixture showed enhanced synergy (CI ranging from 0.18 ± 0.04 to 1.36 ± 0.26). In conclusion, we show for the first time that four major polyphenols from wine synergistically inhibit VSMC proliferation.

Stobadine·2HCl and its two hydrophilic derivatives SM1dM9dM10·2HCl and SME1i- ProC2·HCl were tested in the function of antioxidants on hyaluronan (HA) degradation induced by the Weissberger oxidative system (ascorbate plus Cu(II)). As a primary method, rotational vis- cometry was applied, where the substance tested was added before or 1 h after the initiation of HA degradation. The most effective scavengers ofOH and peroxy-type radicals were recorded to be stobadine·2HCl and SME1i-ProC2·HCl, respectively. The most effective scavenger, determined by applying the ABTS assay, was stobadine·2HCl.

High-molecular-weight hyaluronan (HA) samples were exposed to free-radical chain-degradation reactions induced by ascorbate in the presence of Cu(II) ions – the so-called Weissberger's oxidative system. The concentrations of both reactants [ascorbate, Cu(II)] were comparable to those that may occur during an early stage of the acute phase of joint inflammation. The time-dependent changes of the viscosity of the HA solution in the absence of the substance tested were monitored by rotational viscometry. However, when the anti- or pro-oxidative effects of the antioxidants/drugs were investigated, their dose-dependency was also examined. Additionally, the anti-oxidative activities of these substances were screened by the well-established ABTS and DPPH decolorization assays. The actions of the disease-modifying anti-rheumatic drugs, namely bucillamine and d -penicillamine, were compared to those of l -cysteine and of SA981, the oxidized metabolite of bucillamine. The results indicated that bucillamine was the most efficient scavenger of hydroxyl- and/or peroxyl-type radicals, even at the lowest drug concentration. In contrast, SA981 demonstrated no scavenging activity against the aforementioned free radicals. d -Penicillamine and l -cysteine showed a dual effect, i.e. a pronounced anti-oxidative effect was, after a given time period, followed by a significant pro-oxidative effect.

The aim of the study was a HPLC evaluation of the lipoxygenase activity inhibiting activity of a water infusion of Ligustrum vulgare L. leaves and selected isolates from it. The antiradical activity of the water infusion was determined using DPPH, ABTS and FRAP tests. Oleuropein and echinacoside concentrations in the water infusion were determined by HPLC. Water infusion, echinacoside and oleuropein were used for an antilipoxygenase activity assay using lipoxygenase isolated from rat lung cytosol fraction. Activity of 8-LOX, 12-LOX and 15-LOX were monitored through formation of 8-HETE, 12-HETE and 15-HETE, respectively. The water infusion exhibited the highest activity against all lipoxygenases, followed by oleuropein. Echinacoside was ineffective against LOXs in lower concentrations, while higher concentration showed similar inhibition on 8-LOX and 12-LOX. 15-LOX was affected more and the presence of echinacoside remarkably decreased its activity.

Contribution to the TLC separation of ursolic and oleanolic acid mixtureThe aim of the study was to develop a rapid, simple, effective and reproducible TLC method for separation of a naturally occurring mixture of ursolic and oleanolic acids. Because of the similarity of chemical structures,in situderivatisation by iodine was necessary to separate these triterpenic acids. Separation was achieved on silica gel plates. After derivatisation, a chromatographic plate was developed with the mobile phase consisting of light petrol, ethyl acetate and acetone (8.2:1.8:0.1, v/v/v) following visualisation by spraying with sulphuric acid in diethylether (25%, v/v) and heating to 120°C for 5 min. The method used enabled chromatographical differentiation of ursolic and oleanolic acid mixtures in all tested mutual ratios.

Simple and rapid TLC and column chromatographic methods have been established for separation of luteolin-7-rutinoside and luteolin-7-neohesperidoside on polyamide. A flavonoid mixture was isolated from the butanolic fraction of a methanolic extract of leaves of Ligustrum vulgare L. HPLC analysis of the isolated mixture showed the presence of two compounds in a ratio of approximately 1:1. TLC and column separation was achieved on polyamide with CHCl 3 -MeOH-2-butanone-acetylacetone 9:4:2:1 as mobile phase. After successful separation, the isolated compounds were identified by UV, MS, and NMR spectroscopy and by acid hydrolysis then comparison with authentic samples. Luteolin-7-neohesperidoside was isolated from Ligustrum vulgare L. for the first time.

Hyaluronan (HA) molecules were exposed to free radical-mediated degradation performed by the reaction mixture Cu(II) and ascorbate, the so-called Weissberger biogenic oxidative system, which mimics the situation of acute inflammation. To achieve protection of HA from degradation, herbal extracts such as Agrimonia herba, Cynare folium, and Ligustri folium were selected.Time- and dose-dependent changes of dynamic viscosity of the HA solutions in the presence and absence of the herbal extracts were recorded by rotational viscometry (RV). Radical scavenging capacity of the extracts was investigated by the spectrocolorimetric ABTS and DPPH assays.The results of RV revealed that the extracts of Agrimonia herba and Cynare folium were effective in inhibiting the degradation of HA. On the other hand, the extract of Ligustri folium increased the rate of HA degradation. The highest radical scavenging capacity of ABTS(*+) and DPPH(*) was observed in Agrimonia herba extract followed by the extracts of Ligustri folium and Cynare folium.

To assess protective effects of the tara (Caesalpinia spinosa) extract against hyaluronan (HA) degradation evoked by cupric ions and ascorbate.Uninhibited/inhibited HA degradation was assayed by a decrease in dynamic viscosity of the HA solutions, whereas as a method rotational viscometry was used. To determine radical scavenging capacity of the tara extract, the ABTS and DPPH assays were performed.The results of rotational viscometry showed remarkable protective effects of the tara extract against the degradation of HA. In the ABTS and DPPH assays the IC50 values of the tara extract 1.59 and 30.8 μg/mL indicated quite high radical scavenging properties.The tara extract is an efficient antioxidant as demonstrated by the methods used.

A human transformed line of HeLa cells was treated with extracts prepared from Gymnocladus dioicus, Holodiscus discolor, Stephanandra tanakae, Ligustrum delavayanum, Ligustrum vulgare and Staphylea pinnata. The study evaluating the influence of the active extracts on HeLa cell division also reveals their acute, delayed and combined effect. A 35% degradation of HeLa cells was found after 72 h treatment with 62.5 microg/mL of the extract isolated from Stephanandra tanakae. A 100% lysis of HeLa cells was observed after 72 h treatment with a 125 microg/mL concentration of the extract prepared from Gymnocladus dioicus. The extracts from Ligustrum devayanum and Ligustrum vulgare were specifically effective only with HeLa cells. On the other hand, the extract prepared from Gymnocladus dioicus was effective on the bacteria and on the HeLa cells.

The mutagenicity (bleaching activity) of ofloxacin (43 microM) and acridine orange (AO) (13.5 microM) in Euglena gracilis is inhibited by plant phenolics. Caffeic acid (CA), p-coumaric acid (PCA), ferulic acid (FA) and gentisic acid (GA) (25, 50, 100 and 250 microM) exhibited a significant concentration-dependent inhibitory effect against ofloxacin-induced mutagenicity, which was very effectively eliminated by the highest concentration of all four of those phenolic acids. The mutagenicity of AO was also significantly reduced in the presence of CA, PCA and FA (25, 50, 100 and 250 microM). However, GA exhibited no significant activity, even at the concentration of 250 microM. Based on the UV spectrophotometric measurements, we suggest that the antimutagenic effect of CA, PCA, FA and GA resulted from the scavenging of reactive oxygen species (ROS) produced by ofloxacin. On the other hand, the reduction of AO-induced mutagenicity correlates with the binding capabilities of CA, PCA and FA, with the exception of GA.

Leaf decoctions from Ligustrum vulgare (LV) and Ligustrum delavayanum (LD) were studied for candidacidal activity, phagocytic activity studied on human mononuclear cells (MO) and complement activated by the classical pathway. Candidacidal activity was studied on Candida albicans SC 1539 incubated with MO. The decoction of LD increased the candidacidal activity of MO, whereas LV did not show any effect. The phagocytic activity of MO was decreased by the decoction of LV, whereas LD did not change the activity. The phagocytic index of MO incubated with LD decoction was increased, but use of the LV decoction did not show significant changes. Decoctions from LD and LV significantly decreased the haemolytic activity of complement activated by the classical pathway (conc. 0.78 mg/mL).

The chemical composition of the oils obtained from Thuja occidentalis L. ‘malonyana’ and T. occidentalis ‘malonyana’ (selecton Misak) were determined by GC/MS for the first time. In total 33 components were statistically evaluated by multivariate methods to examine the complex variation of the data set. The main compounds in all oil samples (α-thujone, β -thujone, fenchone) represent more than 50% of the examined oils. Comparison of T. occidentalis ‘malonyana’ and T. occidentalis ‘malonyana’ (selecton Misak) oils showed no substantial differences in the chemical composition. Oil composition was very close to that of T. occidentalis of North American origin.

The antimutagenicity of 14 naturally occurring flavonoids (20 mumol/l) on ofloxacin (43 mumol/l and 86 mumol/l)-induced bleaching (mutagenicity) was studied in Euglena gracilis. The flavonoids chrysin, techtochrysin, chrysin-5-methylether galangin, galangin-5-methylether, pinocembrin and pinobanksin possess considerable antimutagenic properties against ofloxacin-induced bleaching of E. gracilis. Apigenin and isalpinin had only weak antimutagenic potency. Pinobanksin-5-methylether and pinobanksin-3-acetate showed very weak or no antimutagenic effect. However, kempferol, quercetin-3-methylether and quercetin-3,3'-dimethylether showed co-mutagenic or no antimutagenic effect depending on the concentration of ofloxacin. Two possible modes of action of the flavonoids on ofloxacin-induced bleaching of E. gracilis are discussed.

The free radical scavenging activity of the water infusions, different organic solvent extracts and some constituents from Ligustrum vulgare and Ligustrum delavayanum leaves was assessed with the aid of DPPH radical. Among the samples screened, water infusions had the strongest free radical scavenging capacity. From the tested compounds scavenging active flavonoid aglycones are present in the most active chloroform fractions from both leaves samples.

Philadelphus coronarius L. is big, leggy and deciduous old-fashioned shrub known for its fragrant white flowers in the late spring. Some members of genus Philadelphus L. are known for their antibacterial, antiradical and immunomodulatory effects. Therefore, these herbs represent prospective sources for the isolation of active substances with desired effects. We have investigated the cytotoxicity effects of water extracts from leaves and branches of Philadelphus coronarius L. (Hydrangeaceae). A431 cells (human skin carcinoma cell line) and the human breast adenocarcinoma cell line (MCF-7) were treated with various doses of individual extracts (0,1-100 microg dry matter/ml) for 24 h and 72 h. The highest toxic effects of both plant parts extracts were observed on MCF-7 cells regardless the time of treatment. Cells A431 were less sensitive to toxic effects of leaves and branches extracts but the time dependence was present with the tendency of increased toxicity after chronic treatment. There were no differences in the extent of toxic effects between branches and leaves extracts. The results obtained so far will provide the basis for the future studies with isolated active substances from these extracts.

The amyloidoses are diseases associated with nonnative folding of proteins and characterized by the presence of protein amyloid aggregates. The ability of quercetin, resveratrol, caffeic acid, and their equimolar mixtures to affect amyloid aggregation of hen egg white lysozyme in vitro was detected by Thioflavin T fluorescence assay. The anti-amyloid activities of tested polyphenols were evaluated by the median depolymerization concentrations DC50 and median inhibition concentrations IC50 . Single substances are more efficient (by at least one order) in the depolymerization of amyloid aggregates assay than in the inhibition of the amyloid formation with IC50 in 10(-4) to 10(-5) M range. Analyzed mixture samples showed synergic or antagonistic effects in both assays. DC50 values ranged from 10(-5) to 10(-8) M and IC50 from 10(-5) to 10(-9) M, respectively. We observed that certain mixtures of studied polyphenols can synergistically inhibit production of amyloids aggregates and are also effective in depolymerization of the aggregates. Synergic or antagonistic effects of studied mixtures were correlated with protein-small ligand docking studies and AFM results. Differences in these activities could be explained by binding of each polyphenol to a different amino acid sequence within the protein. Our results indicate that synergic/antagonistic anti-amyloid effects of studied mixtures depend on the selective binding of polyphenols to the known amyloidogenic sequences in the lysozyme chain. Our findings of the effective reduction of amyloid aggregation of lysozyme by polyphenol mixtures in vitro are of the utter physiological relevance considering the bioavailability and low toxicity of tested phenols.

The present work summarizes results of isolation and identification of polar constituents of the methanolic extract of Ligustrum vulgare L. leaves and of the evaluation of inhibiting activity of selected isolates on rat lung cytosol fraction lipoxygenase. Six different compounds were isolated from the ethylacetate and butanol portions of the methanolic extract (hydroxytyrosol and its glucoside, ligustroflavon, oleuropein, acteoside, echinacoside). The inhibitory activity of oleuropein, echinacoside and the water infusion of Ligustrum vulgare leaves tested on LOX was expressed as IC50. Kinetic parameters (K M, V max) and type of inhibition were determined. As the most effective in competitive inhibition of LOX, oleuropein was proved.