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Cardiovascular Health and Anti-inflammatory Benefits: In-Vitro

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Bioactivities of pilot-scale extracted cranberry juice and pomace.

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Authors
Harrison JE, Oomah BD, Diarra MS, Ibarra-Alvarado C
Journal
J Food Process Pres 37(4):356-365
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.

Cranberries and wild blueberries treated with gastrointestinal enzymes positively modify glutathione mechanisms in Caco-2 cells in vitro

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Authors
Slemmer JE, Livingston-Thomas JM, Gottschall-Pass KT, Sweeney MI
Journal
J Food Sci 78(6):H943-7
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

Effects of cranberry components on human aggressive periodontitis gingival fibroblasts.

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Authors
Tipton DA, Babu JP, Dabbous MKh
Journal
J Periodontal Res 48(4):433-42
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.

Anti-inflammatory and neuroactive properties of selected fruit extracts

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Authors
Heim KC, Angers P, Léonhart S, Ritz BW
Journal
J Med Food 15(9):851-4
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.

Antioxidant activity of polyphenol rich fruits on human erythrocytes

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Authors
Widen C, Ekholm A, Piwowar-Zali D, Rumpunen K
Journal
Acta Hort 926, 669-674
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.

The antioxidant activity and cytotoxicity methanol extracts from cranberry plants

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Authors
Mustarichie R, Udin Z, Muhtadi A, Surahman E, Subarnas A, Supriyatna
Journal
Int Res J Pharm App Sci
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.

Berry fruits modulated endothelial cell migration and angiogenesis via phosphoinositide-3 kinase/protein kinase B pathway in vitro in endothelial cells.

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Authors
Tulio AZ Jr, Chang C, Edirisinghe I, White KD, Jablonski JE, Banaszewski K, Kangath A, Tadapaneni RK, Burton-Freeman B, Jackson LS
Journal
J Agric Food Chem 60(23):5803-12
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.

Protective effects of the phenolic extracts of fruits against oxidative stress in human lung cells.

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Authors
Boateng J, Verghese M
Journal
Int J Pharmacol 8(3):152-60
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 (p0.05) 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 (p0.05) 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.

Free Radical-Scavenging Properties and Antioxidant Activity of Fractions from Cranberry Products

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Authors
Caillet S, Lorenzo G, Côté J, Sylvain JF, Lacroix M
Journal
Food Nutr Sci 3;337-347
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.

Cellular antioxidant activity (CAA) assay for assessing antioxidants, foods, and dietary supplements.

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Authors
Wolfe KL, Liu RH
Journal
J Agric Food Chem 55(22):8896-907
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.