The Effect of Different Growth Stages of Black Chokeberry Fruits on Phytonutrients, Anti-Lipase Activity, and Antioxidant Capacity.

The present study investigated the nutrients, biologically-active compounds, as well as antioxidant and anti-lipase activities of chokeberry fruits across four different stages of development, from the unripe green to mature black forms. The highest content of total phenolics (12.30% dry weight (DW)), including proanthocyanidins (6.83% DW), phenolic acids (6.57% DW), flavanols (0.56% DW), flavonols (0.62% DW), and flavanones (0.10% DW), was observed in unripe fruits. The unripe green fruits were also characterized by the highest content of protein (2.02% DW), ash (4.05% DW), total fiber (39.43% DW), and chlorophylls (75.48 mg/100 g DW). Ripe black fruits were the richest source of total carotenoids (8.53 mg/100 g DW), total anthocyanins (2.64 g/100 g DW), and total sugars (33.84% DW). The phenolic compounds of green fruits were dominated by phenolic acids (above 83% of the total content), the semi-mature fruits by both phenolic acids and anthocyanins (90%), while the mature berries were dominated by anthocyanins (64%). Unripe fruits were the most effective inhibitor of pancreatic lipase in triolein emulsion, scavenger of 2,2'-azinobis-(3-ethylbenzothiazolin-6-sulfonic acid) radical cation, and reducer of ferric ion. Biological activities were mainly correlated with total proanthocyanidins and total phenolics. Considering their strong anti-lipase and antioxidant activities, unripe chokeberry fruits may have potential applications in nutraceuticals and functional foods.

In Vitro Inhibitory Effects of Viburnum opulus Bark and Flower Extracts on Digestion of Potato Starch and Carbohydrate Hydrolases Activity.

One of the effective treatments for diabetes is to reduce and delay the absorption of glucose by inhibition of α-amylase and α-glucosidase in the digestive tract. Currently, there is a great interest in natural inhibitors from various part of plants. In the present study, the phenolic compounds composition of V. opulus bark and flower, and their inhibitory effects on in vitro potato starch digestion as well as on α-amylase and α-glucosidase, have been studied. Bark and flower phenolic extracts reduced the amount of glucose released from potato starch during tree-stage simulated digestion, with IC50 value equal to 87.77 µg/mL and 148.87 µg/mL, respectively. Phenolic bark extract showed 34.9% and 38.4% more potent inhibitory activity against α-amylase and α-glucosidase, respectively, but the activity of plant extracts was lower than that of acarbose. Chlorogenic acid (27.26% of total phenolics) and (+)-catechin (30.48% of total phenolics) were the most prominent phenolics in the flower and bark extracts, respectively. Procyanidins may be responsible for the strongest V. opulus bark inhibitory activity against α-amylase, while (+)-catechin relative to α-glucosidase. This preliminary study provides the basis of further examination of the suitability of V. opulus bark compounds as components of nutraceuticals and functional foods with antidiabetic activity.

The Effect of Simulated In Vitro Digestion on Biological Activity of Viburnum opulus Fruit Juices.

In the present study, an in vitro digestion method has been used to assay the influence of the physiological conditions in the mouth, stomach, and intestine on the stability and activity in different cell models of the main phenolic compounds from Viburnum opulus fresh juice (FJ), phenolic-rich juice (PJ), and the bioavailable fractions (DFJ and DPJ). The data obtained indicate that the V. opulus samples achieved after in vitro digestion had an influence on cellular glucose and lipid metabolism. The bioavailable fraction of both digested juices stimulated glucose uptake and decreased lipid accumulation by L6 myoblasts and HepG2 hepatocytes. Both DFJ and DPJ reduced the secretion of inflammatory cytokines by 3T3-L1 adipocytes: interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Simultaneously, DFJ and DPJ enhanced oxidative stress in MIN6 cells and decreased glucose-stimulated insulin secretion (GSIS). UPLC-MS analysis revealed qualitative and quantitative changes in hydroxycinnamic acids. In particular, the content of chlorogenic acid decreased drastically; its content in the bioavailable fraction was almost 7 times and 30 times lower than in the FJ and PJ, respectively. Our results suggested that although the phenolic compounds of V. opulus juices undergo transformation during digestion, they are still potent antioxidant agents with biological activity.

An In Vitro Study of the Effect of Viburnum opulus Extracts on Key Processes in the Development of Staphylococcal Infections.

Staphylococcus aureus is still one of the leading causes of both hospital- and community-acquired infections. Due to the very high percentage of drug-resistant strains, the participation of drug-tolerant biofilms in pathological changes, and thus the limited number of effective antibiotics, there is an urgent need to search for alternative methods of prevention or treatment for S. aureus infections. In the present study, biochemically characterized (HPLC/UPLC-QTOF-MS) acetonic, ethanolic, and water extracts from fruits and bark of Viburnum opulus L. were tested in vitro as diet additives that potentially prevent staphylococcal infections. The impacts of V. opulus extracts on sortase A (SrtA) activity (Fluorimetric Assay), staphylococcal protein A (SpA) expression (FITC-labelled specific antibodies), the lipid composition of bacterial cell membranes (LC-MS/MS, GC/MS), and biofilm formation (LIVE/DEAD BacLight) were assessed. The cytotoxicity of V. opulus extracts to the human fibroblast line HFF-1 was also tested (MTT reduction). V. opulus extracts strongly inhibited SrtA activity and SpA expression, caused modifications of S. aureus cell membrane, limited biofilm formation by staphylococci, and were non-cytotoxic. Therefore, they have pro-health potential. Nevertheless, their usefulness as diet supplements that are beneficial for the prevention of staphylococcal infections should be confirmed in animal models in the future.

Viburnum opulus L. Juice Phenolic Compounds Influence Osteogenic Differentiation in Human Osteosarcoma Saos-2 Cells.

Bone mass loss occurs with a decrease in osteoblast proliferation and differentiation, or the enhancement of bone resorption, which further leads to the impairment of bone mineral density and increase in bone fracture. Recent studies suggest that some phenolic compounds found in food play positive role in bone metabolism. High content of phenolic compounds with potential beneficial effects on bone metabolism have been identified in the Viburnum opulus fruit. The aim of the study was to determine the influence of V. opulus fresh juice (FJ) and juice purified by solid phase extraction (PJ) on osteogenesis processes with osteosarcoma Saos-2 cell lines. V. opulus purified juice revealed stronger potential as an inducer of Saos-2 osteogenic differentiation. Saos-2 cells matrix mineralization was evaluated with alkaline phosphatase (ALP) activity measurement and alizarin red S staining. Gene expression analysis showed the elevation of the mRNA levels of Runt-related transcription factor 2 (RUNX2), ALP, collagen type 1 and osteonectin, whereas the nuclear factor-κB ligand and osteoprotegerin ratio (RANKL/OPG) decreased. Furthermore, V. opulus was able to diminish the secretion of pro-inflammatory cytokines Il6 and TNFα, however had no effect on vascular endothelial growth factor (VEGF). It decreased intracellular oxidative stress and induced DNA repair, but had no effect on the growth inhibition of lactic acid beneficial microorganisms.

Molecular Mechanisms of Leonurus Cardiaca L. Extract Activity in Prevention of Staphylococcal Endocarditis-Study on in Vitro and ex Vivo Models.

Better understanding the mechanisms of Leonurus cardiaca L. extract (LCE) activity is necessary to prepare recommendations for the use of LCE-based herbal products for preventive/supportive purposes in case of infective endocarditis (IE) and other staphylococcal invasive infections. The aim of the study was to analyze molecular mechanisms of LCE effect on Staphylococcus aureus and blood platelets in the context of their interactions playing a pivotal role in such disorders. Using atomic force microscopy, we demonstrated that adhesion forces of S. aureus were markedly reduced after exposure to LCE at subinhibitory concentrations. The effect resulted from the impact of LCE on S. aureus cell morphology and the composition of phospholipids and fatty acids in bacterial membranes (assessed by HPLC), which modulated their stabilization, hydrophobicity, and charge. Moreover, using FACS we showed also that LCE significantly reduced GP IIb/IIIa expression on blood platelets, thus the disruption of platelet-fibrinogen interactions seems to explain antiplatelet effect of LCE. The obtained results prove the usefulness of LCE in the prevention of S. aureus adhesion, platelet activation, and vegetations development, however, also pointed out the necessity of excluding the cationic antibiotics from the treatment of S. aureus-associated IE and other invasive diseases, when motherwort herb is used simultaneously as an addition to the daily diet.

Comparison of Chemical Composition and Antioxidant Capacity of Fruit, Flower and Bark of Viburnum opulus.

In this work, the profiles of phenolics, fiber, pectins, sugars, organic acids and carotenoids, vitamin C, ash, protein and fat contents, as well as antioxidant capacity were compared in fruits, flowers, and bark of Viburnum opulus (VO). Antioxidant capacity was evaluated against ABTS, hydroxyl, peroxyl and superoxide free radicals, and as a reducing power by using in vitro test. The results showed great quantitative differences in the composition of the VO morphological parts tested. Fruits contained the highest concentrations of fat, organic acids, sugars, soluble dietary fiber (10.57 ± 0.54; 7.34 ± 0.06; 32.27 ± 1.25; 6.82 ± 0.38 g/100 g DW, respectively) and carotenoids (2.70 ± 0.07 mg/100 g DW). Whereas, the bark exceeded the remaining parts of the VO in terms of antioxidant capacity, ash (9.32 ± 0.17 g/100 g DW), total (59.34 ± 0.75 g/100 g DW) and insoluble dietary fiber (58.20 ± 0.73 g/100 g DW) contents as well as phenolic compounds (3.98 ± 0.04 g/100 g DW). Among the phenolic compounds quantified in this study, chlorogenic acid and (+)-catechin had the highest concentrations (> 1 g/100 g DW) in the flowers and bark, respectively.

Comparison of cytotoxic and anti-platelet activities of polyphenolic extracts from Arnica montana flowers and Juglans regia husks.

Polyphenolic compounds of plant origin are well known to be beneficial to human health: they exert protective effects on haemostasis and have a particular influence on blood platelets. However, the anti-platelet properties of polyphenolic compounds observed so far have not been weighed against their potential cytotoxic action against platelets. The aim of this study was to demonstrate that anti-platelet and cytotoxic effects on blood platelets may interfere and therefore, may often lead to confusion when evaluating the properties of plant extracts or other agents towards blood platelets. The anti-platelet and cytotoxic in vitro effects of plant extracts obtained from the husks of walnuts (J. regia) and flowers of arnica (A. montana) on platelet reactivity and viability were examined. Platelet function was assessed using standard methods (flow cytometry: P-selectin expression, activation of GPIIbIIIa complex, vasodilator-stimulated phosphoprotein, VASP index; turbidimetric and impedance aggregometry) and newly set assays (flow cytometric monitoring of platelet cytotoxicity). The results reveal that none of the studied plant extracts demonstrated cytotoxicity towards blood platelets. The phenolic acid-rich extract of A. montana (7.5 and 15 µg/ml) significantly reduced the ADP-induced aggregation in both whole blood and PRP, and decreased the platelet reactivity index (PRI; VASP phosphorylation) in whole blood, while showing excellent antioxidant capacity. The extract of J. regia husks significantly reduced ADP-induced platelet aggregation in whole blood when applied at 7.5 µg/ml, and only slightly decreased the PRI at 15 µg/ml. Both examined extracts suppressed platelet hyper-reactivity, and such influence did not interfere with cytotoxic effects of the extracts. Thus, its high polyphenol content, excellent antioxidant capacity and distinct anti-platelet properties, in combination with its lack of toxicity, make the extract of A. montana flowers a possible candidate as an anti-platelet agent or a compounding diet supplement.

Effects of Fruit Extracts on Pancreatic Lipase Activity in Lipid Emulsions.

Pancreatic lipase is the most important enzyme in digestion of triglycerides. Reduction of energy intake from dietary fat through inhibition of this enzyme may be a strategy to prevent and treat obesity. In this study, the effect of 31 fruit extracts on pancreatic lipase was investigated using triolein, sunflower oil and rapeseed oil emulsions. Surprisingly, about 30 % of the fruits tested stimulated pancreatic lipase activity in oil emulsions by over 50 %. Only six fruit extracts were found to inhibit pancreatic lipase activity with the IC(50) value varying from 21.11 to 266.48 mg of fruit equivalent/ml of emulsion. Among them, chokeberry demonstrated the highest anti-lipase activity. The inhibitory activity ranks were comparable in all lipid emulsion models and suggest that consumption of chokeberry, red gooseberry and red currant fruits may be a new dietary option for reduction of fat absorption via inhibition of pancreatic lipase.

CD39/NTPDase-1 expression and activity in human umbilical vein endothelial cells are differentially regulated by leaf extracts from Rubus caesius and Rubus idaeus.

Many experimental studies have demonstrated the favorable biological activities of plants belonging to the genus Rubus, but little is known of the role of Rubus leaf extracts in the modulation of the surface membrane expression and activity of endothelial apyrase. The aim of this study was to assess the influence of 1-15 µg/ml Rubus extracts on CD39 expression and enzymatic activity, and on the activation (ICAM-1 expression) and viability of human umbilical vein endothelial cells (HUVEC). The polyphenolic contents and antioxidative capacities of extracts from dewberry (R. caesius L.) and raspberry (R. idaeus L.) leaves were also investigated. The techniques applied were flow cytometry (endothelial surface membrane expression of ICAM-1 and CD39), malachite green assay (CD39 activity), HPLC-DAD (quantitative analysis of polyphenolic extract), ABTS, DPPH and FRAP spectrometric assays (antioxidant capacity), and the MTT test (cell viability). Significantly increased CD39 expressions and significantly decreased ATPDase activities were found in the cells treated with 15 µg/ml of either extract compared to the results for the controls. Neither of the extracts affected cell proliferation, but both significantly augmented endothelial cell ICAM-1 expression. The overall antioxidant capacities of the examined extracts remained relatively high and corresponded well to the determined total polyphenol contents. Overall, the results indicate that under in vitro conditions dewberry and raspberry leaf extracts have unfavorable impact on endothelial cells.

Influence of polyphenol extract from evening primrose (Oenothera paradoxa) seeds on human prostate and breast cancer cell lines.

There is growing interest in plant polyphenols which exhibit pleiotropic biological activities, including anti-inflammatory, antioxidant, and anticancer effects. The objective of our study was to evaluate the influence of an evening primrose extract (EPE) from defatted seeds on viability and invasiveness of three human cell lines: PNT1A (normal prostate cells), DU145 (prostate cancer cells) and MDA-MB-231 (breast cancer cells). The results revealed that after 72 h of incubation the tested extract reduced the viability of DU 145 and MDA-MB-231 with IC50 equal to 14.5 µg/mL for both cell lines. In contrast, EPE did not inhibit the viability of normal prostate cells. Furthermore, EPE reduced PNT1A and MDA-MB-231 cell invasiveness; at the concentration of 21.75 µg/mL the suppression of invasion reached 92% and 47%, respectively (versus control). Additionally, zymographic analysis revealed that after 48 h of incubation EPE inhibited metalloproteinase-2 (MMP-2) and metalloproteinase-9 (MMP-9) activities in a dose-dependent manner. For PNT1A the activities of MMP-2 and MMP-9 decreased 4- and 2-fold, respectively, at EPE concentration of 29 µg/mL. In the case of MDA-MB-231 and DU 145 the decrease in MMP-9 activity at EPE concentration of 29 µg/mL was 5.5-fold and almost 1.9-fold, respectively. In conclusion, this study suggests that EPE may exhibit antimigratory, anti-invasive and antimetastatic potential towards prostate and breast cancer cell lines.

Flavanols from Japanese quince (Chaenomeles japonica) fruit inhibit human prostate and breast cancer cell line invasiveness and cause favorable changes in Bax/Bcl-2 mRNA ratio.

Polyphenols are natural compounds of high structural diversity which translates into a very wide spectrum of biological activities, including chemoprevention. Here we report that a Japanese quince fruit flavanol preparation (JQFFP) caused favorable changes in Bax/Bcl-2 mRNA ratio, which rendered normal and cancer cells more resistant and more sensitive, respectively, to apoptosis. DU145 human prostate cancer cells were characterized by the most advantageous Bax/Bcl-2 ratio. The growth and invasiveness of MDA-MB-231 human breast cancer cells were strongly suppressed by JQFFP, which was accompanied with a decrease in MMP-9 activity and stimulation of TIMP-1 expression. Importantly, JQFFP did not decrease normal human prostate PNT1A cell number, whereas Bax/Bcl-2 ratio decreased which implies increased resistance to apoptosis. In conclusion, JQFFP exhibited a potent antiproliferative effect against cancer cells, inhibited their invasiveness, and decreased expression level of several genes involved in apoptosis, angiogenesis, and metastasis.

Procyanidins from evening primrose (Oenothera paradoxa) defatted seeds inhibit invasiveness of breast cancer cells and modulate the expression of selected genes involved in angiogenesis, metastasis, and apoptosis.

There is a growing interest in plant polyphenols (including flavanols) that exhibit pleiotropic biological activities such as antiinflammatory, antioxidant, and anticancer effects. Here, we report for the first time the inhibition of MDA-MB-231 breast cancer cell viability and invasiveness by an evening primrose flavanol preparation (EPFP). We observed a decrease in MDA-MB-231 viability of 50% vs. a control after 72 h of incubation with EPFP at a concentration of 58 µM gallic acid equivalents (GAE) and an inhibition of their invasiveness of 65% vs. a control at 75 µM GAE after 48 h of incubation. EPFP caused a 10-fold reduction in matrix metalloproteinase-9 (MMP-9) activity at 100 µM GAE. Furthermore, through modulation of mRNA expression, EPFP reduced the expression levels of the following proteins: antiapoptotic Bcl-2, angiogenic vascular endothelial growth factor (VEGF), and 2 transcription factors (c-Jun, c-Fos). Moreover, analysis by flow cytometry revealed that EPFP induced apoptosis in MDA-MB-231 cells. In conclusion, our data shows that EPFP inhibits cell viability by increasing apoptosis and decreases cell invasiveness by decreasing angiogenesis.

Influence of phytocenosis on the medical potential of moss extracts: the Pleurozium schreberi (Willd. ex Brid.) Mitt. case.

The question was asked "whether plant phytocenosis has an impact on the medical potential of the extracts from Pleurozium schreberi". Moss samples were collected from four different phytocoenoses: mixed forest (oak-pine forest), a forest tract in pine forest, 5-15-year-old pine forest and 50-year-old pine forest. Chemical composition of the extracts, antioxidative capacity (FRAP and ABTS·+ assays), as well as biological activities including cytotoxicity for the mouse fibroblasts L929 line (MTT reduction assay), biostatic/biocidal effect against selected bacteria and fungi (broth microdilution method followed by culture on solid media), and regenerative properties on human fibroblasts HFF-1 line (scratch assay) were tested. The conducted research clearly proves that phytocenosis determines the quality of moss extracts. The analyses showed that in every examined aspect the IV-7 extract (obtained from a specimen collected in a Pinus sylvestris L. forest, monoculture up to 15 years old) exhibited the highest values and the strongest activity. Other extracts of the same species but growing in other phytocenoses-in a mixed forest (IV-5), a forest tract in a Pinus sylvestris monoculture forest (IV-6) and in a P. sylvestris forest of pine monoculture about 50 years old (IV-8) showed much weaker activity and lower values of the above-mentioned parameters. At the same time, none of the tested extracts exerted a pro-regenerative effect. The P. schreberi extracts were characterized by a varied total content of phenolic compounds in the range from 0.63 ± 0.02 to 14.01 ± 0.25 mg/g of plant material. UPLC/MS analysis showed a varied phenolic profile of the extracts, with caffeoylquinic acid and quercetin triglucoside predominating in all of them.

Morphological and Photosynthetic Parameters of Green and Red Kale Microgreens Cultivated under Different Light Spectra.

Microgreens are plants eaten at a very early stage of development, having a very high nutritional value. Among a large group of species, those from the Brassicaceae family, including kale, are very popularly grown as microgreens. Typically, microgreens are grown under controlled conditions under light-emitting diodes (LEDs). However, the effect of light on the quality of grown microgreens varies. The present study aimed to determine the effect of artificial white light with varying proportions of red (R) and blue (B) light on the morphological and photosynthetic parameters of kale microgreens with green and red leaves. The R:B ratios were for white light (W) 0.63, for red-enhanced white light (W + R) 0.75, and for white and blue light (W + B) 0.38 at 230 µmol m-2 s-1 PPFD. The addition of both blue and red light had a positive effect on the content of active compounds in the plants, including flavonoids and carotenoids. Red light had a stronger effect on the seedling area and the dry mass and relative chlorophyll content of red-leaved kale microgreens. Blue light, in turn, had a stronger effect on green kale, including dry mass. The W + B light combination negatively affected the chlorophyll content of both cultivars although the leaves were significantly thicker compared to cultivation under W + R light. In general, the cultivar with red leaves had less sensitivity to the photosynthetic apparatus to the spectrum used. The changes in PSII were much smaller in red kale compared to green kale. Too much red light caused a deterioration in the PSII vitality index in green kale. Red and green kale require an individual spectrum with different proportions of blue and red light at different growth stages to achieve plants with a large leaf area and high nutritional value.

Hypolipidemic and antioxidant effects of hydroxycinnamic acids, quercetin, and cyanidin 3-glucoside in hypercholesterolemic erythrocytes (in vitro study).

OBJECTIVE: Four polyphenols: ferulic acid and p-coumaric acid (hydroxycinnamic acids), quercetin (flavonol) and cyanidin 3-glucoside (anthocyanin) were selected, and their antioxidant properties and their influence on cholesterol concentration in hypercholesterolemic and normal erythrocytes were investigated. METHODS: To determine the effect of phenolic compounds, we prospectively studied cholesterol concentration, lipid peroxidation and membranes fluidity. Whole-blood and isolated erythrocytes (2% hematocrit) were incubated for 24 h with selected compounds at concentration 1, 10 and 100 µmol/L. All investigated compounds decreased lipid peroxidation in whole blood. Cyanidin 3-glucoside and quercetin showed higher antioxidant properties than hydroxycinnamic acids (ferulic acid and p-coumaric acid). RESULTS: Incubation of whole blood of hypercholesterolemic patients with quercetin and cyanidin 3-glucoside resulted in statistically significant reduction of cholesterol concentration in erythrocytes down to 75% (at 10 µmol/L of polyphenols) and 69% (at 100 µmol/L of polyphenols) of initial values. The effect of both compounds on isolated erythrocytes was even more pronounced, reduction down to 70% (at 10 µmol/L of polyphenols) and 58% (at 100 µmol/L of polyphenols) of initial values. After incubation of isolated erythrocytes of hypercholesterolemic patients with quercetin and cyanidin 3-glucoside, increase of membrane fluidity was noticed. After incubation of isolated erythrocytes of healthy donors with investigated compounds, no changes in membrane fluidity were observed. CONCLUSION: Our results indicate that flavonols and anthocyanins have higher antioxidant properties and higher influence on cholesterol concentration in erythrocytes membranes than simple hydroxycinnamic acids.

Proanthocyanidins as the main pancreatic lipase inhibitors in chokeberry fruits.

Pancreatic lipase inhibitors are recognized as important in strategies for the management of overweight and obesity. The phytocompounds in chokeberry fruit show multidirectional pro-health effects, including anti-obesity activity. The aims of this study were to fractionate and identify the phenolic compounds of chokeberry fruit phenolic-rich extract that are active as pancreatic lipase inhibitors. Phenolic compounds were fractionated using Sephadex LH-20 resin, followed by polyphenol profile analysis using chromatographic and spectrophotometric methods, while pancreatic inhibitory activity was determined using 4-methylumbelliferyl oleate and emulsified triolein as enzyme substrates. Among the six fractions isolated from extract, two fractions rich in highly polymerized proanthocyanidins showed the greatest ability to inhibit pancreatic lipase activity. In comparison, fractions containing mainly low-molecular-weight phenolic compounds, such as phenolic acids, flavonols and anthocyanins, were 11-64 times less active. The most active fraction showed a mixed mode of pancreatic lipase inhibition, as determined by Lineweaver-Burk plot analysis, and exhibited a cumulative effect with orlistat. This study shows that black chokeberry polyphenols, particularly highly polymerized procyanidins, can effectively inhibit pancreatic lipase activity determined by in vitro methods.

Procyanidins from Japanese quince (Chaenomeles japonica) fruit induce apoptosis in human colon cancer Caco-2 cells in a degree of polymerization-dependent manner.

Plant proanthocyanidins, including procyanidins, display various biological activities. Here we report an inhibition of human colon cancer Caco-2 cell growth by the extract from Japanese quince fruit and the procyanidin-rich fractions of the extract. We observed that the amount of apoptotic Caco-2 cells increased by 52.1% vs. control after 72-h incubation with 50 µg extract/mL, as assessed by flow cytometry and image cytometry. Under the same experimental conditions the corresponding values for human colon cancer HT-29 cells and for rat normal intestinal IEC-6 cells were 5.0% and 8.1%, respectively. The extract fractions enriched with higher oligomers exhibited the highest proapoptotic activity. In conclusion, the Japanese quince procyanidins exhibited proapoptotic activity in Caco-2 cells within a submilimolar concentration range.

Glycoside Hydrolases and Non-Enzymatic Glycation Inhibitory Potential of Viburnum opulus L. Fruit-In Vitro Studies.

Phytochemicals of various origins are of great interest for their antidiabetic potential. In the present study, the inhibitory effects against carbohydrate digestive enzymes and non-enzymatic glycation, antioxidant capacity, and phenolic compounds composition of Viburnum opulus L. fruits have been studied. Crude extract (CE), purified extract (PE), and ethyl acetate (PEAF) and water (PEWF) fractions of PE were used in enzymatic assays to evaluate their inhibitory potential against α-amylase with potato and rice starch as substrate, α-glucosidase using maltose and sucrose as substrate, the antioxidant capacity (ABTS, ORAC and FRAP assays), antiglycation (BSA-fructose and BSA-glucose model) properties. Among four tested samples, PEAF not only had the highest content of total phenolics, but also possessed the strongest α-glucosidase inhibition, antiglycation and antioxidant activities. UPLC analysis revealed that this fraction contained mainly chlorogenic acid, proanthocyanidin oligomers and flavalignans. Contrary, the anti-amylase activity of V. opulus fruits probably occurs due to the presence of proanthocyanidin polymers and chlorogenic acids, especially dicaffeoylquinic acids present in PEWF. All V. opulus samples have an uncompetitive and mixed type inhibition against α-amylase and α-glucosidase, respectively. Considering strong anti-glucosidase, antioxidant and antiglycation activities, V. opulus fruits may find promising applications in nutraceuticals and functional foods with antidiabetic activity.

Characteristics of the Polyphenolic Profile and Antioxidant Activity of Cone Extracts from Conifers Determined Using Electrochemical and Spectrophotometric Methods.

The aim of the study was to analyze the polyphenolic profile of cone extracts of Douglas fir, Scots pine and Korean fir, and to study their antioxidant activity. The mechanism of electro-oxidation of polyphenols (such as procyanidins and catechins) from cone extracts was investigated using cyclic voltammetry (CV) and differential pulse voltammetry (DPV), as well as spectrophotometric methods-ABTS (2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate)), DPPH (2,2-diphenyl-1-picrylhydrazyl), FRAP (Ferric Reducing Antioxidant Power ) and CUPRAC (CUPric Reducing Antioxidant Capacity). The scientific novelty of the research is the comprehensive analysis of cone extracts in terms of antioxidant properties. Due to the high polyphenol content, the extracts showed significant ability to reduce oxidative reactions, as well as the ability to scavenge free radicals and transition metal ions. Douglas fir, Scots pine and Korean fir cone extracts can potentially be used as natural stabilizers, preservatives and antimicrobial substances in the food industry and in medications.