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Cardiovascular Health & Anti-inflammatory Benefits: In-Vitro

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MALDI-TOF MS characterization of proanthocyanidins from cranberry fruit (Vaccinium macrocarpon) that inhibit tumor cell growth and matrix metalloproteinase expression in vitro.

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Authors
Neto CC, Krueger CG, Lamoureaux TL, Kondo M, Vaisberg AJ, Hurta RAR, Curtis S, et al
Journal
J Sci Food Agr 86(1):18-25
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

Potential role of dietary flavonoids in reducing microvascular endothelium vulnerability to oxidative and inflammatory insults

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Authors
Youdim KA, McDonald J, Kalt W, Joseph JA
Journal
J Nutr Biochem 13(5):282-288
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

Regulation of vascular endothelial function by procyanidin-rich foods and beverages

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Authors
Caton PW, Pothecary MR, Lees DM, Khan NQ, Wood EG, Shoji T, Kanda T, Rull G and Corder R
Journal
J Agric Food Chem 58(7):4008-13
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.

Antioxidant levels of common fruits, vegetables, and juices versus protective activity against in vitro ischemia/reperfusion.

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Authors
Bean H, Schuler C, Leggett RE and Levin RM
Journal
Int Urol Nephrol 42(2):409-15
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.

Composition of a chemopreventive proanthocyanidin-rich fraction from cranberry fruits responsible for the inhibition of 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced ornithine decarboxylase (ODC) activity.

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Authors
Kandil FE, Smith MA, Rogers RB, Pépin MF, Song LL, Pezzuto JM, Seigler DS
Journal
J Agric Food Chem 50(5):1063-9
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.

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)

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Authors
Kahlon TS, Smith GE
Journal
Food Chem 100(3):1182-1187
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 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).

Cranberry extract inhibits low density lipoprotein oxidation

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Authors
Wilson T, Porcari JP, Harbin D
Journal
Life Sci 62(24):A381-6
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.

Potential of cranberry-based herbal synergies for diabetes and hypertension management

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Authors
Apostolidis E, Kwon YI, Shetty K
Journal
Asia Pac J Clin Nutr 15(3):433-41
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.

Fruit extracts antagonize A beta or DA-induced deficits in Ca2+ flux in M1-transfected COS-7 cells

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Authors
Joseph JA, Fisher DR, Carey AN
Journal
JAD 6(4):403-411
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.

Anti-angiogenic property of edible berries

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Authors
Roy S, Khanna S, Alessio HM, Vider J, Bagchi D, Bagchi M, Sen CK
Journal
Free Radic Res 36(9):1023-31
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.