Abstract: Cranberry juice has been long used to prevent infections because of its effect on the adhesion of the bacteria to the host surface. Proanthocyanidins (PACs) comprise of one of the major classes of phytochemicals found in cranberry, which have been extensively studied and found effective in combating adhesion of pathogenic bacteria. The role of other cranberry constituents in impacting bacterial adhesion haven't been studied very well. In this study, cranberry juice fractions were prepared, characterized and tested for their effect on the surface adhesion of the pathogenic clinical bacterial strain E. coli B78 and non-pathogenic control E. coli HB101. The preparations tested included crude cranberry juice extract (CCE); three fractions containing flavonoid classes including proanthocyanidins, anthocyanins and flavonols; selected sub-fractions, and commercially available flavonol glycoside, quercetin-3-O-galactoside. Atomic force microscopy (AFM) was used to quantify the adhesion forces between the bacterial surface and the AFM probe after the treatment with the cranberry fractions. Adhesion forces of the non-pathogenic, non fimbriated lab strain HB101 are small (average force 0.19 nN) and do not change with cranberry treatments, whereas the adhesion forces of the pathogenic, Dr adhesion E. coli strain B78 (average force of 0.42 nN) show a significant decrease when treated with cranberry juice extract or fractions (average force of 0.31 nN, 0.37 nN and 0.39 nN with CCE, Fraction 7 and Fraction 4 respectively). In particular, the fractions that contained flavonols in addition to PACs were more efficient at lowering the force of adhesion (average force of 0.31 nN-0.18 nN between different sub-fractions containing flavonols and PACs). The sub-fractions containing flavonol glycosides (from juice, fruit and commercial quercetin) all resulted in reduced adhesion of the pathogenic bacteria to the model probe. This strongly suggests the anti adhesive role of other classes of cranberry compounds in conjunction with already known PACs and may have implications for development of alternative anti bacterial treatments.
Abstract: The protective effect of proanthocyanidin-containing polyphenol extracts from apples, avocados, cranberries, grapes, or proanthocyanidin microbial metabolites was evaluated in colonic epithelial cells exposed to p-cresol, a deleterious compound produced by the colonic microbiota from L-tyrosine. In HT29 Glc-/+ cells, p-cresol significantly increased LDH leakage and decreased ATP contents, whereas in Caco-2 cell monolayers, it significantly decreased the transepithelial electrical resistance and increased the paracellular transport of FITC-dextran. The alterations induced by p-cresol in HT29 Glc-/+ cells were prevented by the extracts from cranberries and avocados, whereas they became worse by extracts from apples and grapes. The proanthocyanidin bacterial metabolites decreased LDH leakage, ameliorating cell viability without improving intracellular ATP. All of the polyphenol extracts and proanthocyanidin bacterial metabolites prevented the p-cresol-induced alterations of barrier function. These results suggest that proanthocyanidin-containing polyphenol extracts and proanthocyanidin metabolites likely contribute to the protection of the colonic mucosa against the deleterious effects of p-cresol.
Abstract: The preventive effects of the American cranberry (Vaccinium macrocarpon) against urinary tract infections are supported by extensive studies which have primarily focused on its phenolic constituents. Herein, a phenolic-free carbohydrate fraction (designated cranf1b-F2) was purified from cranberry fruit using ion exchange and size exclusion chromatography. MALDI-TOF-MS analysis revealed that the cranf1b-F2 constituents are predominantly oligosaccharides possessing various degrees of polymerisation and further structural analysis (by GC-MS and NMR) revealed mainly xyloglucan and arabinan residues. In antimicrobial assays, cranf1b-F2 (at 1.25 mg/mL concentration) reduced biofilm production by the uropathogenic Escherichia coli CFT073 strain by over 50% but did not inhibit bacterial growth. Cranf1b-F2 (ranging from 0.625 to 10 mg/mL) also inhibited biofilm formation of the non-pathogenic E. coli MG1655 strain up to 60% in a concentration-dependent manner. These results suggest that cranberry oligosaccharides, in addition to its phenolic constituents, may play a role in its preventive effects against urinary tract infections.
Abstract: A novel methodology was developed to elucidate proanthocyanidins (PAC) interaction with extra-intestinal pathogenic Escherichia coli (ExPEC). PAC inhibit ExPEC invasion of epithelial cells and, therefore, may prevent transient gut colonization, conferring protection against subsequent extra-intestinal infections, such as urinary tract infections. Until now PAC have not been chemically labeled with fluorophores. In this work, cranberry PAC were labeled with 5-([4,6-dichlorotriazin-2-yl]amino) fluorescein (DTAF), detected by high-performance liquid chromatography with diode-array detection and characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). We report single and double fluorescent-labeled PAC with one or two chlorine atoms displaced from DTAF in alkaline pH via nucleophilic substitution. Fluorescent labeling was confirmed by fragmentation experiments using MALDI-TOF/TOF MS. Fluorescent labeled PAC were able to promote ExPEC agglutination when observed with fluorescence microscopy. DTAF tagged PAC may be used to trace the fate of PAC after they agglutinate ExPEC and follow PAC-ExPEC complexes in cell culture assays.
Abstract: Respiratory viruses are a major public health problem because of their prevalence and high morbidity rate leading to considerable social and economic implications. Cranberry has therapeutic potential attributed to a comprehensive list of phytochemicals including anthocyanins, flavonols, and unique A-type proanthocyanidins. Soy flavonoids, including isoflavones, have demonstrated anti-viral effects in vitro and in vivo. Recently, it was demonstrated that edible proteins can efficiently sorb and concentrate cranberry polyphenols, including anthocyanins and proanthocyanins, providing greatly stabilized matrices suitable for food products. The combination of cranberry and soy phytoactives may be an effective dietary anti-viral resource. Anti-viral properties of both cranberry juice-enriched and cranberry pomace polyphenol-enriched soy protein isolate (CB-SPI and CBP-SPI) were tested against influenza viruses (H7N1, H5N3, H3N2), Newcastle disease virus and Sendai virus in vitro and in ovo. In our experiments, preincubation with CB-SPI or CBP-SPI resulted in inhibition of virus adsorption to chicken red blood cells and reduction in virus nucleic acid content up to 16-fold, however, CB-SPI and CBP-SPI did not affect hemagglutination. Additionally, CB-SPI and CBP-SPI inhibited viral replication and infectivity more effectively than the commercially available anti-viral drug Amizon. Results suggest CB-SPI and CBP-SPI may have preventative and therapeutic potential against viral infections that cause diseases of the respiratory and gastro-intestinal tract.
Abstract: In this study, we have assessed the phenolic metabolism of a cranberry extract by microbiota obtained from the ascending colon and descending colon compartments of a dynamic gastrointestinal simulator (SHIME). For comparison, parallel fermentations with a grape seed extract were carried out. Extracts were used directly without previous intestinal digestion. Among the 60 phenolic compounds targeted, our results confirmed the formation of phenylacetic, phenylpropionic and benzoic acids as well as phenols such as catechol and its derivatives from the action of colonic microbiota on cranberry polyphenols. Benzoic acid (38.4mug/ml), 4-hydroxy-5-(3'-hydroxyphenyl)-valeric acid (26.2mug/ml) and phenylacetic acid (19.5mug/ml) reached the highest concentrations. Under the same conditions, microbial degradation of grape seed polyphenols took place to a lesser extent compared to cranberry polyphenols, which was consistent with the more pronounced antimicrobial effect observed for the grape seed polyphenols, particularly against Bacteroides, Prevotella and Blautia coccoides-Eubacterium rectale.
Abstract: OBJECTIVE: In the context of increasing microbial resistance and limited new antimicrobials, we aimed to study the antimicrobial effects of cranberry proanthocyanidin extracts on Escherichia coli growth, adhesion to epithelial cells, and lung infection.
DESIGN: Experimental in vitro and in vivo investigation.
SETTING: University research laboratory.
SUBJECTS: Seventy-eight 6- to 8-week-old male Balb/C mice.
INTERVENTIONS: In vitro, the effect of increasing concentrations of cranberry proanthocyanidin on bacterial growth of different clinical E. coli isolates was evaluated. Ex vivo, adhesion of E. coli to fresh human buccal epithelial cells was measured in the presence or absence of cranberry proanthocyanidin using microscopy. In vivo, lung bacterial count, pulmonary immune response (neutrophil murine chemokine keratinocyte-derived cytokine measurement and polymorphonuclear recruitment in bronchoalveolar lavage fluid), and lethality were evaluated in a pneumonia mouse model with E. coli precultured with or without cranberry proanthocyanidin. E. coli isolates originated from ventilated ICU patients with respiratory tract colonization or ventilator- associated pneumonia. They differed in number of virulence genes.
MEASUREMENTS AND MAIN RESULTS: A significant inhibition of bacterial growth was observed with increasing concentration of cranberry proanthocyanidin, affecting both time to maximal growth and maximal growth rate (p<0.0001 for both). The minimal concentration at which this effect occurred was 250 mug/mL. Cranberry proanthocyanidin significantly reduced E. coli adhesion to fresh buccal epithelial cells by up to 80% (p<0.001). Bacterial counts in homogenized lungs and bronchoalveolar lavage fluid were decreased after cranberry proanthocyanidin exposition (p<0.05 and p<0.01, respectively). Cranberry proanthocyanidin also decreased KC concentrations and polymorphonuclear cell recruitment in bronchoalveolar lavage fluid (p<0.05 for both). At identical inoculum, mortality was reduced by more than half in mice inoculated with E. coli exposed to cranberry proanthocyanidin (p<0.01).
CONCLUSION: Cranberry proanthocyanidins exhibit potent effects on growth, adhesion, and virulence of oropharyngeal and lung isolates of E. coli, suggesting that cranberry proanthocyanidin could be of clinical interest to reduce oropharyngeal colonization and prevent lung infection.
Abstract: Cranberry flavonoids (flavonols and flavan-3-ols), in addition to their antioxidant properties, have been shown to possess potential in vitro activity against several cancers. However, the difficulty of isolating cranberry compounds has largely limited anticancer research to crude fractions without well-defined compound composition. In this study, individual cranberry flavonoids were isolated to the highest purity achieved so far using gravity and high performance column chromatography and LC-MS characterization. MTS assay indicated differential cell viability reduction of SKOV-3 and OVCAR-8 ovarian cancer cells treated with individual cranberry flavonoids. Treatment with quercetin aglycone and PAC DP-9, which exhibited the strongest activity, induced apoptosis, led to caspase-3 activation and PARP deactivation, and increased sensitivity to cisplatin. Furthermore, immunofluorescence microscopy and western blot study revealed reduced expression and activation of epidermal growth factor receptor (EGFR) in PAC DP-9 treated SKOV-3 cells. In addition, quercetin aglycone and PAC DP-9 deactivated MAPK-ERK pathway, induced downregulation of cyclin D1, DNA-PK, phospho-histone H3 and upregulation of p21, and arrested cell cycle progression. Overall, this study demonstrates promising in vitro cytotoxic and anti-proliferative properties of two newly characterized cranberry flavonoids, quercetin aglycone and PAC DP-9, against ovarian cancer cells.
Abstract: BACKGROUND AND OBJECTIVE: In periodontitis, gingival epithelial cells can produce interleukin (IL)-6, a regulator of osteoclastic bone resorption, in response to IL-1beta. IL-1beta regulates cytokine expression via signaling pathways, including nuclear factor (NF)-B and mitogen activated protein kinase (MAPK)/activator protein (AP)-1. Cranberry proanthocyanidins (PACs) inhibit IL-1beta-stimulated IL-6 production, but specific mechanisms are unclear. The objectives of this study were to determine effects of cranberry PACs on NF-B and MAPK/AP-1 activation of IL-1beta-stimulated IL-6 production in gingival epithelial cells.
MATERIAL AND METHODS: Cranberry high molecular weight non-dialyzable material (NDM), rich in PACs, was derived from cranberry juice. Human gingival epithelial cells [Smulow-Glickman (S-G)] were incubated with IL-1beta in the presence or absence of NDM or inhibitors of NF-B, [nemo-binding domain (NBD) peptide] or AP-1 (SP600125), and IL-6 levels were measured by ELISA. Effects of NDM on IL-1beta-activated NF-B and AP-1 and phosphorylated intermediates in both pathways were measured in cell extracts via binding to specific oligonucleotides and specific sandwich ELISAs, respectively. Data were analyzed using ANOVA and Scheffe's F procedure for post hoc comparisons.
RESULTS: IL-1beta (> 0.1 nm) caused a time- and dose-dependent stimulation of S-G epithelial cell IL-6 production (p < 0.005). This was significantly decreased in a dose-dependent manner by NBD peptide or SP600125 [maximum inhibition ~30-40% (p < 0.02)], and together, the two inhibitors decreased IL-6 by ~80%, similar to the inhibition caused by NDM (p < 0.001). IL-1beta stimulated NF-B and AP-1 activation (p < 0.003), which was inhibited by NDM (p < 0.0001). NDM did not significantly affect IL-1beta-stimulated levels of phosphorylated intermediates in the NF-B pathway (IBalpha) or the AP-1 pathway (c-Jun, ERK1/2).
CONCLUSION: In S-G epithelial cells, IL-1beta appeared to upregulate IL-6 production via activation of both NF-B and MAPK/AP-1 signaling pathways because cranberry NDM decreased nuclear levels of IL-1beta-activated NF-B (p65) and AP-1 (phospho-c-Jun) and strongly inhibited IL-6 production. Lack of inhibition of phosphorylation of IBalpha, c-Jun or ERK1/2 suggested that NDM might affect both pathways downstream from those points in S-G cells, such as ubiquitination and proteosomal degradation of IBalpha, or inhibition of nuclear activity of c-Jun and/or ERK1/2. Defining these points of inhibition precisely may help identify molecular targets of cranberry polyphenols. 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Abstract: In the context of dental caries prevention by natural foodstuff sources, antifungal and antibiofilm activities of dry commercial extracts of cranberry fruit (Vaccinium macrocarpon Aiton) and two other red fruits (Vaccinium myrtillus L. and Malpighia punicifolia L.) were assessed on Candida albicans and Candida glabrata yeasts. When added to the culture medium, the cranberry extract displayed a significant anti-adhesion activity against Candida spp. when used at low concentrations. In addition, the pretreatment of surfaces with this extract induced an anti-adhesion activity mainly against C. glabrata yeasts and an antibiofilm activity against C. albicans. This activity was dependent on concentration, species, and strain. A phytochemical investigation bioguided by anti-adhesion tests against the two Candida species was carried out on crude cranberry juice to determine the active fractions. Three subfractions enriched in proanthocyanidins showed an anti-adhesion activity at low concentrations. This study investigated for the first time the interest of crude extracts of cranberry and cranberry juice fractions to prevent biofilms of C. glabrata. It highlighted the potency of consuming this fruit and using it as a source of anti-adhesion agents. 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.
Abstract: BACKGROUND AND OBJECTIVE: Gingival epithelial cells and fibroblasts participate in periodontal inflammation and destruction, producing interleukin (IL)-6, a regulator of osteoclastic bone resorption, and the neutrophil chemoattractant IL-8. IL-17, a product of T-helper 17 cells, may play a role in periodontitis by stimulating cytokine production by gingival cells. The cranberry (Vaccinium macrocarpon) is rich in polyphenols, particularly proanthocyanidins, which have antioxidant and other beneficial properties. Cranberry components inhibit pro-inflammatory activities of lipopolysaccharide-stimulated human macrophages, gingival fibroblasts, and epithelial cells, but little is known of its effects on IL-17-stimulated cytokine production. The objectives were to determine the effects of IL-17 + cranberry components on IL-6 and IL-8 production by human gingival epithelial cells and fibroblasts.
MATERIAL AND METHODS: Cranberry high molecular weight non-dialyzable material (NDM), which is rich in proanthocyanidins, was derived from cranberry juice. Human gingival epithelial cells and normal human gingival fibroblasts were incubated with NDM (5-50 mug/mL), IL-17 (0.5-100 ng/mL), or NDM + IL-17 in serum-free medium for 6 d. IL-6 and IL-8 in culture supernatants were measured by ELISA. Membrane damage and viability were assessed by lactate dehydrogenase activity released into cell supernatants and activity of a mitochondrial enzyme, respectively. Data were analyzed using ANOVA and Scheffe's F procedure for post hoc comparisons.
RESULTS: In both cell lines, IL-17 (> ~5-10 ng/mL) significantly stimulated production of IL-6 (p < 0.005) and IL-8 (p < 0.03). Non-toxic levels of NDM inhibited constitutive IL-6 and IL-8 production by epithelial cells (p < 0.01) and fibroblasts (p < 0.03) as well as IL-17-stimulated cytokine production by epithelial cells [IL-6 (maximum ~80% inhibition; p < 0.0001); IL-8 (maximum ~70% inhibition; p < 0.03)] and fibroblasts [IL-6 (maximum ~90% inhibition; p < 0.0001); IL-8 (maximum ~80% inhibition; p < 0.008)].
CONCLUSION: Cranberry NDM inhibition of constitutive and IL-17-stimulated IL-6 and IL-8 production by gingival fibroblasts and epithelial cells suggests that cranberry components could be useful as a host modulatory therapeutic agent to prevent or treat periodontitis. 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Abstract: BACKGROUND: Berries are known to have many kinds of biological activities. We focused on their antiviral effect, which has not yet been well evaluated.
RESULTS: We compared the anti-influenza viral effects of berries belonging to the genus Vaccinium - 35 species of blueberry (Vaccinium cyanococcus), the Natsuhaze (Vaccinium oldhamii), bilberry (Vaccinium myrtillus) and cranberry (Vaccinium oxycoccos)- with those belonging to the genus Ribes, i.e. blackcurrant (Ribes nigrum). Only Elliott and Legacy among Northern Highbush varieties but many Rabbiteye varieties such as Austin, Baldwin, Brightblue, Festival, T-100 and Tifblue showed anti-influenza viral activity. Natsuhaze, bilberry, cranberry and blackcurrant had high antiviral effects. A relationship was observed between the antiviral effect and total polyphenol content.
CONCLUSIONS: Antiviral effects were found to differ markedly between berry species. Rabbiteye varieties tended to have higher antiviral effects than Northern, Southern and Half Highbush blueberry varieties. We also found that Natsuhaze, which has recently been harvested in Japan as a potential functional food, had an antiviral effect comparable to that of bilberry, cranberry and blackcurrant. There was a positive relationship between antiviral activity and polyphenol content, indicating the possibility that polyphenol is one of the key factors in the antiviral effects of berries. 2012 Society of Chemical Industry.
Abstract: Fresh cranberries were processed by two pilot-scale methods to recover juice and extracts from cranberry pomace. Press cake was extracted with three successive ethanol soaks followed by decanting in trial 1 versus one ethanol soak and solvent removal by decanting and compressing with the bladder press in trial 2. Yields and recoveries of juice, dry juice solids, press cake, press cake extractives (PCEs), polyphenolics and antioxidant capacity were determined relative to the input material of fresh cranberries or press cake. PCEs from both processes exhibited strong dose-dependant vasorelaxant effects on rat aorta rings with EC50 of 2.3-3.9 micro g/mL and Emax of 96-98%. PCEs contained three to four times the phenolic acids, tartaric esters and antioxidant activities plus five to 10 times the flavonols and anthocyanins of their respective juice powders. The polyphenolic levels were 121-142, 7-10, 9-11 and 10-19 mg equivalents of catechin, caffeic acid, quercetin and cyanidin-3-glucoside/g of extract, respectively. Antioxidant activities of the PCEs and juices were 201-296 and 64-75 mg trolox equivalents/g powder. Juice yields of 47-58% accounted for only 18-50% of the bioactives recovered from whole fruit. Sequential extraction of the press cake with 95% ethanol and removal of the extract with the bladder press favored high recoveries of polyphenolics with increased antioxidant and vasorelaxant benefits.
Abstract: Beneficial health effects of cranberries (CBs) and wild blueberries (BBs), such as reduced levels of oxidative stress, have been demonstrated in feeding studies. These Vaccinium berries contain high levels of flavonoids; however, the bioavailability of flavonoids is generally low. We investigated the in vitro effects of these berries on intestinal cells, focusing on mitigating oxidative stress and associated reactive oxygen species (ROS). First, we simulated the passage of CB and BB through the gastrointestinal (GI) tract by treating berry homogenates to a battery of digestive enzymes. Then, Caco-2 cells, a model of small intestine epithelial uptake, were exposed to these homogenates for 60 min. Using a cell-free assay, we found that the antioxidant activity in CB homogenates was not affected by these enzymes, but that BB homogenates treated with gut enzymes had 43% lower free-radical quenching activity (P < 0.05). However, both of the enzyme-treated homogenates were still able to counteract the ROS-generating ability of H2O2 added exogenously to Caco-2 cells. Berry homogenates also increased mitochondrial metabolic rates at 60 min posttreatment, as measured by MTT assays. Enzyme-treated CB (but not BB) homogenates increased the levels of reduced glutathione (GSH) relative to oxidized glutathione (GSSG), a critical indicator of the cellular redox state (P < 0.05). Our data suggest that CBs do not lose their antioxidant ability when passing through the GI tract, and specifically, digested CB may serve to enhance cytoprotective effects in intestinal cells by reducing potential damage caused by free radicals and ROS derived from other food sources
Abstract: BACKGROUND AND OBJECTIVE: Aggressive periodontitis (AgP) causes rapid periodontal breakdown involving AgP gingival fibroblast production of cytokines [i.e. interleukin (IL)-6, a bone metabolism regulator], and matrix metalloproteinase (MMP)-3. Lipopolysaccharide upregulates fibroblast IL-6 and MMP-3, via transcription factors (i.e. NF-kB). Cranberry (Vaccinium macrocarpon) inhibits lipopolysaccharide-stimulated macrophage and normal gingival fibroblast activities, but little is known of its effects on AgP fibroblasts. Objectives of this study are to use AgP fibroblasts, to determine cytotoxicity of cranberry components or periodontopathogen (Fusobacterium nucleatum, Porphyromonas gingivalis) lipopolysaccharide +/- cranberry components, and effects of cranberry components on lipopolysaccharide-stimulated NF-kB activation and IL-6 and MMP-3 production.
MATERIAL AND METHODS: AgP fibroblasts were incubated <= 6 d with high molecular weight non-dialyzable material (NDM) (derived from cranberry juice (1-500 mug/mL) or lipopolysaccharide (1 mug/mL) +/- NDM. Membrane damage and viability were assessed by enzyme activity released into cell supernatants and activity of a mitochondrial enzyme, respectively. Secreted IL-6 and MMP-3 were measured by ELISA. NF-kB p65 was measured via binding to an oligonucleotide containing the NF-kB consensus site. Data were analyzed using analysis of variance and Scheffe's F procedure for post hoc comparisons.
RESULTS: Short-term exposure to NDM, or lipopolysaccharide +/- NDM caused no membrane damage. NDM (<= 100 mug/mL) or lipopolysaccharide +/- NDM had no effect on viability <= 7 d exposure. NDM (50 mug/mL) inhibited lipopolysaccharide-stimulated p65 (P <= 0.003) and constitutive or lipopolysaccharide-stimulated MMP-3 (P <= 0.02). NDM increased AgP fibroblast constitutive or lipopolysaccharide-stimulated IL-6 (P <= 0.0001), but inhibited normal human gingival fibroblast IL-6 (P <= 0.01).
CONCLUSION: Lack of toxicity of low NDM concentrations, and its inhibition of NF-kB and MMP-3, suggest that cranberry components may regulate AgP fibroblast inflammatory responses. Distinct effects of NDM on AgP and gingival fibroblast production of IL-6 (which can have both positive and negative effects on bone metabolism) may reflect phenotypic differences in IL-6 regulation in the two cell types.
Abstract: Epidemiological evidence supports inverse associations between fruit and vegetable intake and incidence of
cardiovascular disease and neurodegeneration. Dietary botanicals with salient health benefits include berries and leafy vegetables. Molecular pharmacology research has ascribed these benefits primarily to phenolic constituents and antioxidant activity. The current investigation sought to eluicidate pharmacologic activity of two novel preparations of berry and spinach extracts in vitro. Blueberry and cranberry exhibited the greatest antioxidant activity. In a dose-dependent manner, a proprietary mixture of cranberry and blueberry extracts inhibited inhibitor of jB kinase b, a central node in inflammatory signal transduction. A proprietary mixture of blueberry, strawberry, and spinach extracts inhibited prolyl endopeptidase, a regulator
of central neuropeptide stability and an emerging therapeutic target in neurology and psychiatry. These results indicate specific molecular targets of blended dietary plants with potential relevance to inflammation and neurological health.
Abstract: Diets rich in fruit and vegetables promote health and delay the onset of diseases associated with oxidative stress. The benefit, especially of different berries, has been largely attributed to their content of numerous phytochemicals, and their effects in terms of antioxidant capacity are often evaluated chemically by different methods. We have instead used a highly relevant biological model, a modified CAP-e assay (Cell-based Antioxidant Protection in erythrocytes), to evaluate bioefficacy of antioxidants in Swedish berries. Extracts of twelve fruit and berries were analysed both by chemical and biological analyses: apple (Malus domestica, peel), bilberry (Vaccinium myrtillus), black currant (Ribes nigrum), purple chokeberry (Aronia x prunifolia), cranberry (Vaccinium macrocarpon), elderberry (Sambucus nigra), lingonberry (Vaccinium vitis-idaea), raspberry (Rubus idaeus), rose hips (Rosa spp.), sea buckthorn (Hippohae rhamnoides), sloe (Prunus spinosa) and strawberry (Fragaria x ananassa). Purple chokeberry, sloe and rose hips showed high antioxidant capacity in the chemical assays. Rose hips showed the highest degree of antioxidant protection also in the biological model, however, chokeberry and sloe showed medium or low protection. Furthermore, strawberry showed overall high protection in the biological assay but low antioxidant capacity in the chemical assays. The chemical and biological models showed different results and future studies of the biological model and in vivo situations are necessary.
Abstract: The purpose of this study was to determine the total phenol content, antioxidant activity and cytotoxicity of methanol extracts from cranberry plants. The highest total phenol content of 17.1 mg/100 g, and antioxidant activity with IC50=23.8 mg/100 g. This situation shows that the total content of phenolic plant extracts examined correlated with DPPH activity. IC50 cytotoxicity of methanol extracts of each 75.11 micro g/mL against Calu-6 cells, 177.53 from micro g/mL against MCF-cells and 54.87 micro g/mL against HCT-116 cells. From the data obtained we can conclude that this plant has a quite high of total phenolic content and antioxidant activity. Correlation between total phenolics increased DPPH free radical scavenging and cytotoxic activities are quite good. The results of this study showed that cranberry plants can be used as the basis for the treatment of some diseases.
Abstract: Polyphenolic-rich berry fruits are known to activate redox-sensitive cellular signaling molecules such as phosphatidylinositol-3-kinase (PI3 kinase)/kinase B (Akt), resulting in a cascade of downstream signaling pathways. This study investigated the ability of strawberry (SB), wild blueberry (WBB), and cranberry (CB) extracts to induce the activation of PI3 kinase/Akt signaling in vitro in human umbilical endothelial cells (HUVECs) and whether this activation would enhance cell migration and angiogenesis. Anthocyanin profiles of the extracts were characterized using HPLC-ESI/MS, and Akt activation was investigated using the Alpha Screen SureFire assay. The total anthocyanin contents of SB, WBB, and CB extracts were 81.7, 82.5, and 83.0 mg/100 g fresh weight, respectively. SB, WBB, and CB extracts activated Akt in a dose-dependent manner via PI3 kinase and induced cell migration and angiogenesis in vitro in HUVECs. The results from this study suggest that polyphenolics in berry fruits may play a role in promoting vascular health.
Abstract: Consumption of fruits and the other dietary antioxidants are considered beneficial due to the protection they afford in the pathogenesis associated with oxidative stress. The aim of this study was to evaluate the antioxidative effects of selected fruit extracts (Plums, Apples, Grapes and Cranberries) on human lung fibroblasts (CCD-25LU) exposed to tert-butyl hydroperoxide (tBHP) oxidative stress. Lactate Dehydrogenase (LDH) was used to assess cytotoxicity (cell integrity) and antioxidant enzymes catalase (CAT), glutathione-s-transferase (GST), glutathione peroxidases (GPx) and concentrations of reduced glutathione (GSH) were determined. Results showed that LDH release by cells pretreated with fruits extracts were significantly (p<0.05) lower compared to cells treated with tBHP alone. Antioxidant enzymes (CAT, GST and GPx) in cells pretreated with fruit extracts were increased by 2-4 folds compared to cell exposed to tBHP alone. GSH levels which were significantly (p<0.05) reduced after exposure to tBHP were restored by pretreatment with fruit extracts. Fruits extracts used in this study protected CCD-25LU against oxidative stress induced by tBHP and reduced cell damage. Consumption of fruits may therefore play a significant role in protection against oxidative induced lung diseases.
Abstract: Lipid peroxidation inhibition capacity and antiradical activity were evaluated in HPLC fractions of different polarity obtained from two cranberry juices and three extracts isolated from frozen cranberries and pomace containing antho- cyanins, water-soluble and apolar phenolic compounds, respectively. Compounds with close polarities were collected to obtain between three and four fractions from each juice or extract. The cranberry phenols are good free radical-scav- engers, but they were less efficient at inhibiting the lipid peroxidation. Of all the samples tested, the intermediate pola- rity fraction of extract rich in apolar phenolic compounds of fruit presented the highest antiradical activity while the most hydrophobic fractions of the anthocyanin-rich extract from fruit and pomace appeared to be the most efficient at inhibiting the lipid peroxidation. The antioxidant or pro-oxidant activity of fractions increased with the concentration. The phenol polarity and the technological process to manufacture cranberry juice can influence the antioxidant and an- tiradical activities of fractions.
Abstract: A cellular antioxidant activity (CAA) assay for quantifying the antioxidant activity of phytochemicals, food extracts, and dietary supplements has been developed. Dichlorofluorescin is a probe that is trapped within cells and is easily oxidized to fluorescent dichlorofluorescein (DCF). The method measures the ability of compounds to prevent the formation of DCF by 2,2′-azobis(2-amidinopropane)
dihydrochloride (ABAP)-generated peroxyl radicals in human hepatocarcinoma HepG2 cells. The decrease in cellular fluorescence when compared to the control cells indicates the antioxidant capacity of the compounds. The antioxidant activities of selected phytochemicals and fruit extracts were evaluated using the CAA assay, and the results were expressed in micromoles of quercetin equivalents per 100 μmol of phytochemical or micromoles of quercetin equivalents per 100 g of fresh fruit. Quercetin had the highest CAA value, followed by kaempferol, epigallocatechin gallate (EGCG), myricetin, and luteolin among the pure compounds tested. Among the selected fruits tested, blueberry had the highest CAA value, followed by cranberry > apple ) red grape > green grape. The CAA
assay is a more biologically relevant method than the popular chemistry antioxidant activity assays because it accounts for some aspects of uptake, metabolism, and location of antioxidant compounds within cells.
Abstract: Chitosan binds to negatively charged soy lecithin liposomes by an electrostatic interaction driven by its positively charged amino group. This interaction allows stable covered vesicles (chitosomes) to be developed as a suitable targeted carrier and controlled release system. This study investigated the effect of chitosomes on the activation of cranberry proanthocyanidins (PAC) in Raw 264.7 macrophages. Chitosomes were characterized according to size, zeta potential, PAC-loading, and release properties. Results showed an increase in the net positive charge and size of the liposomes as the concentration of chitosan was increased, suggesting an effective covering of the vesicles by means of electrostatic interactions, as shown by transmission electron microscopy and fluorescence microscopy. About 85% of the PAC that was loaded remained in the chitosomes after release studies for 4 hours in phosphate-buffered saline. Cyclo-oxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are associated with inflammation. Activated RAW 264.7 macrophages increase the expression of COX-2 and iNOS in response to bacterial infection and inflammation; we, therefore, tested the ability of the PAC-loaded chitosomes to attenuate COX-2 and iNOS expression in LPS (lipopolysaccharide)-stimulated macrophages. Increasing the amount of PAC loaded into the chitosomes caused a dose-dependent attenuation of iNOS and COX-2 expression in LPS-stimulated macrophages. A 2% v/v PAC-loaded chitosomes formulation almost completely attenuated the LPS-induced expression of iNOS and COX-2. PAC-loaded chitosomes were more active than PAC alone, suggesting that the macrophage response to LPS occurs after endocytosis of the PAC-loaded chitosomes.
Abstract: Flavonoids can bind the divalent cations frequently used to evaluate LDL antioxidant capacity in vitro. Flavonoids in cranberry juice (CBJ) may serve as antioxidants and promote cardiovascular health. This in vitro study characterizes CBJ effects on metal and non-metal based oxidation of human LDL. For cupric ion-initiated oxidation of LDL, thiobarbituric acid reactive substances (TBARS) formation and relative electrophoretic mobility (REM) were significantly inhibited by CBJ at a dilution of 1:10,000. Diene formation during LDL oxidation was evaluated by continuous measurement of absorbance at 234 nm. The time required for cupric ion-initiated LDL oxidations to reach maximum reaction velocity was significantly delayed by 1:10,000 dilutions of CBJ. Non-metal initiated LDL oxidation by 2,2'-azobis-amidinopropane was significantly inhibited by CBJ at dilutions of 1:10,000 and 1:5,000 for REM and TBARS tests, respectively. Protection of LDL from both metal and non-metal based oxidative injury confirms that the effects of CBJ are not due to flavonoid chelation of oxidants but due to a true and potent antioxidant capacity.
Abstract: Abstract: Antioxidant activity of six fractions of cranberry phenolic compounds was determined by
inhibition of Cu2+-induced low-density lipoprotein (LDL) oxidation. The phenolic composition of each fraction was determined by high-performance liquid chromatography. The phenolic fractions were mixed with aliquots of modified human serum prior to LDL isolation. The serum was modified to remove very-low-density lipoprotein and chylomicrons that may bind phenolic compounds. Only fractions 5 and 6 that contained proanthocyanidins (PAs) significantly increased the lag time of LDL oxidation, and the lag time for fraction 6 was significantly higher than for fraction 5. The mass distribution of PAs in these fractions was obtained by matrix-assisted laser desorption/ionisation time- of-flight mass spectrometry, a technique that allows rapid characterisation of the molecular weight distribution in mixtures of oligomeric compounds. Fraction 5 contained trimers through heptamers, whereas fraction 6 contained pentamers through nonamers. In addition, fraction 6 contained PA oligomers with more doubly linked, A-type interflavan bonds. Results indicate that PAs specifically associate with LDL in modified serum and increase the lag time of Cu2+-induced oxidation. Differences between fractions 5 and 6 in PA structure and effects on LDL oxidation suggest that the degree of polymerisation and the nature of the interflavan bond influence antioxidant properties
Abstract: Abstract:Proanthocyanidin-rich extracts were prepared by fractionation of the fruit of theNorthAmerican
cranberry (Vaccinium macrocarpon). In vitro growth inhibition assays in eight tumor cell lines showed
that selected fractions inhibited the growth of H460 lung tumors, HT-29 colon and K562 leukemia cells at
GI50 values ranging from 20 to 80 μgml−1. Matrix-assisted laser desorption/ionization time-of-flight mass
spectrometry (MALDI-TOF MS) of one of these fractions found it to be composed of polyflavan-3-ols,
which are primarily tetramers through heptamers of epicatechin containing one or two A-type linkages.
Whole cranberry extract and the proanthocyanidin fractions were screened for effect on the expression of
matrix metalloproteinases in DU 145 prostate carcinoma cells. The expression of MMP-2 and MMP-9 was
inhibited in response to whole cranberry extract and to a lesser degree by the proanthocyanidin fractions
Abstract: Although antioxidant systems help control the level of reactive oxygen species they may be overwhelmed during periods of oxidative stress. Evidence suggests that oxidative stress components as well as inflammatory mediators may be involved in the pathogenesis of vascular disorders, where localized markers of oxidative damage have been found. In this regard we investigated the putative antioxidant and anti-inflammatory effects of blueberry and cranberry anthocyanins and hydroxycinnamic acids against H2O2 and TNF induced damage to human microvascular endothelial cells. Polyphenols from both berries were able to localize into endothelial cells subsequently reducing
endothelial cells vulnerability to increased oxidative stress at both the membrane and cytosol level. Furthermore, berry polyphenols also reduced TNF induced up-regulation of various inflammatory mediators (IL-8, MCP-1 and ICAM-1) involved in the recruitment of leukocytes to sites of damage or inflammation along the endothelium. In conclusion, polyphenols isolated from both blueberry and cranberry were able to afford protection to endothelial cells against stressor induced up-regulation of oxidative and inflammatory insults. This may have beneficial actions against the initiation and development of vascular diseases and be a contributing factor in the reduction of age-related
deficits in neurological impairments previously reported by us
Abstract: Flavonoid-rich diets are associated with a lower mortality from cardiovascular disease. This has been linked to improvements in endothelial function. However, the specific flavonoids, or biologically active metabolites, conferring these beneficial effects have yet to be fully defined. In this experimental study of the effect of flavonoids on endothelial function cultured endothelial cells have been used as a bioassay with endothelin-1 (ET-1) synthesis being measured an index of the response.
Evaluation of the relative effects of extracts of cranberry juice compared to apple, cocoa, red wine, and green tea showed inhibition of ET-1 synthesis was dependent primarily on their oligomeric procyanidin content. Procyanidin-rich extracts of cranberry juice triggered morphological changes in endothelial cells with reorganization of the actin cytoskeleton and increased immunostaining for phosphotyrosine residues. These actions were independent of antioxidant activity. Comparison of the effects of apple procyanidin monomers through heptamer showed a clear structure-activity
relationship. Although monomer, dimer, and trimer had little effect on ET-1 synthesis, procyanidin tetramer, pentamer, hexamer, and heptamer produced concentration-dependent decreases with IC50 values of 5.4, 1.6, 0.9, and 0.7 μM, respectively. Levels of ET-1 mRNA showed a similar
pattern of decreases, which were inversely correlated with increased expression of Kruppel-like factor 2 (KLF2), a key endothelial transcription factor with a broad range of antiatherosclerotic actions including suppression of ET-1 synthesis. Future investigations of procyanidin-rich products should assess the role KLF2 induction plays in the beneficial vascular effects of high flavonoid consumption.
Abstract: It is well known that antioxidants present in various fruits, vegetables, and juices have the potential to protect the urinary bladder from free radical damage. What is not well understood, however, is how well antioxidant activities detected by chemical methods such as the CUPRAC assay for total antioxidant activity (TAA) predict the level of physiological protection available. It is hypothesized that the level of antioxidant reactivity found by the CUPRAC assay will positively correlate with increased protection in a model of in vitro ischemia/reperfusion. To test this hypothesis, the CUPRAC assay was utilized to determine the antioxidant reactivity of a series of fruits, vegetables, and juices, and the results were compared to the protective ability of selected juices in an established in vitro rabbit bladder model of ischemia/reperfusion. The results of the CUPRAC test showed that cranberry juice had the highest level of antioxidant reactivity, blueberry juice had an intermediate activity, and orange juice had the lowest. It was determined, however, that contrary to the hypothesis, the orange juice was significantly more potent in protecting the bladder against ischemia/reperfusion damage than either blueberry or cranberry juice. Thus, it is concluded that chemical tests for TAA do not necessarily correlate with their physiological activity.
Abstract: Phenolics from the American cranberry (Vaccinium macrocarpon) were fractionated into a series of proanthocyanidins and other flavonoid compounds by vacuum chromatography on a hydrophilic, porous polyvinylic gel permeation polymer. Antioxidant activity was not restricted to a particular class of components in the extract but was found in a wide range of the fractions. Significant chemopreventive activity, as indicated by an ornithine decarboxylase assay, was localized in one particular proanthocyanidin-rich fraction from the initial fractionation procedure. Further fractionation of the active anticarcinogenic fraction revealed the following components: seven flavonoids, mainly quercetin, myricetin, the corresponding 3-O-glycosides, (-)-epicatechin, (+)-catechin, and dimers of both gallocatechin and epigallocatechin types, and a series of oligomeric proanthocyanidins.
Abstract: The in vitro binding of bile acids by blueberries (Vaccinium spp.), plums (Prunus spp.), prunes (Prunus spp.), strawberries (Fragaria x ananassa), cherries (Malpighia punicifolia) cranberries (Vaccinium macrocarpon) and apples (Malus sylvestris) was determined using a mixture of bile acids secreted in human bile at a duodenal physiological pH of 6.3. Six treatments and two blank incubations were conducted to testing various fresh raw fruits on an equal dry matter basis. Considering cholestyramine (bile acid binding, cholesterol lowering drug) as 100% bound, the relative in vitro bile acid binding on dry matter (DM), total dietary fibre (TDF) and total polysaccharides (PCH) basis was for blueberries 7%, 47% and 25%; plums 6%, 53% and 50%; prunes 5%, 50% and 14%; strawberries 5%, 23% and 15%; cherries 5%, 37% and 5%; cranberries 4%, 12% and 7%; and apple 1%, 7% and 5%, respectively. Bile acid binding on DM basis for blueberries was significantly (P<=0.05) higher than all the fruits tested. The bile acid binding for plums was similar to that for prunes and strawberries and significantly higher than cherries, cranberries and apples. Binding values for cherries and cranberries were significantly higher than those for apples. These results point to the relative health promoting potential of blueberries > plums=prunes=strawberries=cherries=cranberries > apples as indicated by their bile acid binding on DM basis. The variability in bile acid binding between the fruits tested maybe related to their phytonutrients (antioxidants, polyphenols, hydroxycinnamic acids, flavonoids, anthocyanins, flavonols, proanthocyanidins, catechins), structure, hydrophobicity of undigested fractions, anionic or cationic nature of the metabolites produced during digestion or their interaction with active binding sites. Inclusion of blueberries, plums, prunes, strawberries, cherries and cranberries in our daily diet as health promoting fruits should be encouraged. Animal studies are planned to validate in vitro bile acid binding of fruits observed herein to their potential of atherosclerosis amelioration (lipid and lipoprotein lowering) and cancer prevention (excretion of toxic metabolites).
Abstract: Cranberry juice consumption is often used for the treatment of urinary tract infections, but the effect of cranberry juice on heart disease has not been investigated. We evaluated how a cranberry extract containing 1,548 mg gallic acid equivalents/liter (initial pH=2.50) affected low density lipoprotein (LDL) oxidation induced by 10 micromolar cupric sulfate. When LDL oxidation took place in the presence of diluted cranberry extracts, the formation of thiobarbituric acid reactive substances (TBARS) and LDL electrophoretic mobility were reduced. LDL electrophoretic migration was also reduced when the cranberry extract had a pH of 7.00 prior to dilution. This study suggests that cranberry extracts have the ability to inhibit the oxidative modification of LDL particles.
Abstract: Water soluble cranberry-based phytochemical combinations with oregano, rosemary, and Rhodiola rosea were evaluated for total phenolic content, related antioxidant activity and inhibition of diabetes management-related alpha -glucosidase, pancreatic alpha-amylase inhibition, and hypertension-related ACE-I inhibitory activities. Water extracts of oregano had 114.9 mg/g DW of phenolics which was highest among all the extracts tested, whereas the 75% cranberry with 25% oregano combinations had the highest phenolics (38.9 mg/g DW) among all the combinations tested. The water extracts of oregano had the highest DPPH radical inhibition activity (73.6 %), whereas among combinations the 75% cranberry and 25% oregano had the highest DPPH radical inhibition activity (50.8 %). These results indicated a correlation between total phenolic content and antioxidant activity. The water extracts of pure Rhodiola rosea had the highest alpha -glucosidase inhibition, whereas the 75% cranberry and 25% Rhodiola rosea combination had the highest inhibition among the combinations. In the case of alpha -amylase inhibition the water extracts of Rhodiola rosea had the highest inhibition, whereas the 75% cranberry with 25% Rhodiola rosea combination had the highest inhibition among the combinations. All the water extracts tested indicated that they had anti-ACE-I inhibitory activity. More specifically, among the water extracts 100% cranberry had the highest ACE-I inhibitory activity and among the combination the 75% cranberry with 25% rosemary had the highest ACE-I inhibitory activity. The analysis of alpha -glucosidase,alpha -amylase, and ACE-I inhibitory activities suggested that inhibition depend on the phenolic profile of each unique extract and by bringing together synergistic combinations to cranberry, health beneficial functionality was enhanced. This enhanced functionality in terms of high alpha -glucosidase and alpha -amylase inhibitory activities indicate the potential for diabetes management, and high ACE-I inhibitory activity indicates the potential for hypertension management.
Abstract: Evidence suggests that there is a selective sensitivity to oxidative stress (OSS) among muscarinic receptor (MAChR) subtypes with M1, M2 and M4 showing > OSS than M3 or M5 subtypes in transfected COS-7 cells. This may be important in determining the regional specificity in neuronal aging and Alzheimer disease (AD). We assessed the effectiveness of blueberry (BB) and other high antioxidant (HA) fruit extracts (boysenberry, BY; cranberry, CB; black currant, BC; strawberry, SB; dried plums, DP; and grape, GR) on the toxic effects of Abeta 25-35 (100 microM, 24 hrs) and DA (1 mM, 4 hrs) on calcium buffering (Recovery) following oxotremorine (750 microM) -induced depolarization in M1AChR-transfected COS-7 cells, and on cell viability following DA (4 hrs) exposure. The extracts showed differential levels of Recovery protection in comparisons to the non-supplemented controls that was dependent upon whether DA or Abeta was used as the pretreatment. Interestingly, assessments of DA-induced decrements in viability revealed that all of the extracts had some protective effects. These findings suggest that the putative toxic effects of Abeta or DA might be reduced by HA fruit extracts.
Abstract: Recent studies show that edible berries may have potent chemopreventive properties. Anti-angiogenic approaches to prevent and treat cancer represent a priority area in investigative tumor biology. Vascular endothelial growth factor (VEGF) plays a crucial role for the vascularization of tumors. The vasculature in adult skin remains normally quiescent. However, skin retains the capacity for brisk initiation of angiogenesis during inflammatory skin diseases such as psoriasis and skin cancers. We sought to test the effects of multiple berry extracts on inducible VEGF expression by human HaCaT keratinocytes. Six berry extracts (wild blueberry, bilberry, cranberry, elderberry, raspberry seed, and strawberry) and a grape seed proanthocyanidin extract (GSPE) were studied. The extracts and uptake of their constituents by HaCaT were studied using a multi-channel HPLC-CoulArray approach. Antioxidant activity of the extracts was determined by ORAC. Cranberry, elderberry and raspberry seed samples were observed to possess comparable ORAC values. The antioxidant capacity of these samples was significantly lower than that of the other samples studied. The ORAC values of strawberry powder and GSPE were higher than cranberry, elderberry or raspberry seed but significantly lower than the other samples studied. Wild bilberry and blueberry extracts possessed the highest ORAC values. Each of the berry samples studied significantly inhibited both H2O2 as well as TNF alpha induced VEGF expression by the human keratinocytes. This effect was not shared by other antioxidants such as alpha-tocopherol or GSPE but was commonly shared by pure flavonoids. Matrigel assay using human dermal microvascular endothelial cells showed that edible berries impair angiogenesis.
Abstract: Consumption of fruits and vegetables has been associated with reduced risk of chronic diseases such as cardiovascular disease and cancer. Phytochemicals, especially phenolics, in fruits and vegetables are suggested to be the major bioactive compounds for the health benefits. However, the phenolic contents and their antioxidant activities in fruits and vegetables were underestimated in the literature, because bound phenolics were not included. This study was designed to investigate the profiles of total phenolics, including both soluble free and bound forms in common fruits, by applying solvent extraction, base digestion, and solid-phase extraction methods. Cranberry had the highest total phenolic content, followed by apple, red grape, strawberry, pineapple, banana, peach, lemon, orange, pear, and grapefruit. Total antioxidant activity was measured using the TOSC assay. Cranberry had the highest total antioxidant activity (177.0 +/- 4.3 micromol of vitamin C equiv/g of fruit), followed by apple, red grape, strawberry, peach, lemon, pear, banana, orange, grapefruit, and pineapple. Antiproliferation activities were also studied in vitro using HepG(2) human liver-cancer cells, and cranberry showed the highest inhibitory effect with an EC(50) of 14.5 +/- 0.5 mg/mL, followed by lemon, apple, strawberry, red grape, banana, grapefruit, and peach. A bioactivity index (BI) for dietary cancer prevention is proposed to provide a new alternative biomarker for future epidemiological studies in dietary cancer prevention and health promotion.
Abstract: Cardiovascular disease (CVD) is the leading cause of death in most industrialized countries. Cranberries were evaluated for their potential roles in dietary prevention of CVD. Cranberry extracts were found to have potent antioxidant capacity preventing in vitro LDL oxidation with increasing delay and suppression of LDL oxidation in a dose-dependent manner. The antioxidant activity of 100 g cranberries against LDL oxidation was equivalent to 1000 mg vitamin C or 3700 mg vitamin E. Cranberry extracts also significantly induced expression of hepatic LDL receptors and increased intracellular uptake of cholesterol in HepG2 cells in vitro in a dose-dependent manner. This suggests that cranberries could enhance clearance of excessive plasma cholesterol in circulation. We propose that additive or synergistic effects of phytochemicals in cranberries are responsible for the inhibition of LDL oxidation, the induced expression of LDL receptors, and the increased uptake of cholesterol in hepatocytes.