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Cardiovascular Health & Anti-inflammatory Benefits: In-Vitro

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Anti-inflammatory and neuroactive properties of selected fruit extracts

Posted
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.

Posted
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.

Posted
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 (p

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.

Chitosomes loaded with cranberry proanthocyanidins attenuate the bacterial lipopolysaccharide-induced expression of iNOS and COX-2 in raw 264.7 macrophages

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Authors
Madrigal-Carballo S, Rodriguez G, Sibaja M, Reed JD, Vila AO, Molina F
Journal
J Liposome Res 19(3):189-96
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.

Cranberry juice inhibits metal and non-metal initiated oxidation of human low density lipoproteins in vitro

Posted
Authors
Wilson T, Porcari JP, Maher MA
Journal
J Nutraceut Function Med Foods 2(2):5-14
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.

Cranberry proanthocyanidins associate with low-density lipoprotein and inhibit Cu2+ -induced oxidation

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Authors
Porter ML, Krueger CG, Wiebe DA, Cunningham DG, Reed JD
Journal
J Sci Food Agr 81(14):1306-1313
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