Increasing antioxidant activity and reducing decay of blueberries by essential oils.

Several naturally occurring essential oils including carvacrol, anethole, cinnamaldehyde, cinnamic acid, perillaldehyde, linalool, and p-cymene were evaluated for their effectiveness in reducing decay and increasing antioxidant levels and activities in 'Duke' blueberries ( Vaccinium corymbosum). Carvacrol, anethole, and perillaldehyde showed the capability to promote total anthocyanins and total phenolics and enhance antioxidant activity in fruit tissues expressed as oxygen radical absorbance capacity (ORAC) and hydroxyl radical ( (*)OH) scavenging capacity. All of the essential oils tested in this study were able to inhibit fruit decay development to some degree compared to controls. The most effective compound for mold retardation was p-cymene, followed by linalool, carvacrol, anethole, and perillaldehyde. Cinnamic acid and cinnamaldehyde also suppressed mold growth, but to a lesser extent. Treatment with carvacrol, anethole, or perillaldehyde also significantly increased the levels of fructose, glucose, and citric acid. Individual flavonoids were variably affected by the essential oils. Levels of chlorogenic acid, which was the major phenolic compound in blueberry fruit, were enhanced by all of the essential oils in this study. Increased amounts of quercetin 3-galactoside and quercetin 3-arabinoside were also found in all treated fruit except samples treated with linalool or p-cymene. The major anthocyanin, malvidin 3-galactoside, was enhanced by all essential oils tested except linalool and p-cymene. The levels of other individual anthocyanins including petunidin 3-galactoside, delphinidin 3-galactoside, petunidin 3-glucoside, petunidin 3-arabinoside, delphinidin 3-arabinoside, and cyanidin 3-galactoside were higher in treated fruit compared to controls. Those essential oils that have positive effects on enhancing anthocyanins, phenolic compounds, and antioxidant activity of fruit, but inhibitory effects on microbial growth and decay development, deserve further evaluation.

Resveratrol content in strawberry fruit is affected by preharvest conditions.

This study investigated the occurrence of resveratrol in Fragaria x ananassa Duchesne and the effect of preharvest conditions on resveratrol content. Both cis- and trans-resveratrol were detected in strawberry achenes (seeds) and pulp (receptacle tissue). Resveratrol was found to be higher in achenes than in fruit pulp. The levels of resveratrol were affected by genotype variations, fruit maturation, cultural practices, and environmental conditions. High growing temperature (25 and 30 degrees C) or enriched CO 2 in the atmosphere significantly enhanced resveratrol content of strawberries. Advancing maturation also increased resveratrol content. The mature pulp and achenes contained higher amounts of resveratrol than the immature fruit. Adding compost as a soil supplement or preharvest application of methyl jasmonate (MJ) also significantly enhanced the level of resveratrol in strawberry fruit. Among the plants grown in hill plasticulture, fruits of 'Ovation (B28)', 'Mohawwk', 'Earliglow', and 'B35' had higher amounts of resveratrol than fruits of other genotypes. 'Ovation' contained the highest amount of resveratrol among strawberries grown in matted row, whereas 'Latestar' contained the least. Ten of 14 tested genotypes (all except 'Allstar', 'Delmarvel', 'Northeaster', and 'MEUS 8') had higher amounts of resveratrol when grown in hill plasticulture compared to matted row.

Enhancing antioxidant, antiproliferation, and free radical scavenging activities in strawberries with essential oils.

Several natural antimicrobial compounds derived from essential oils of plants were investigated for their efficacies in inhibiting decay and extending the shelf life of strawberries (Fragaria x ananassas Duch.). The severity of decay in strawberries stored at 10 degrees C was significantly reduced by treatment with thymol. Treatments with menthol or eugenol also suppressed the fungal growth, but to a lesser extent. All of these three natural antimicrobial compounds extended shelf life of strawberries as compared to the control. Strawberries treated with thymol, menthol, or eugenol also maintained better fruit quality with higher levels of sugars, organic acids, phenolics, anthocyanins, flavonoids, and oxygen radical absorbance capacity than the untreated fruits. The free radical scavenging properties of strawberry fruit were evaluated against 2,2-diphenyl-1-picryhydrazyl (DPPH(.)), hydroxyl (HO(.)), and superoxide radicals (O2(.-)) using electron spin resonance measurements. Higher radical scavenging capacities were found against DPPH(.) and HO(.) in all treated fruit, particularly in berries treated with thymol, compared to those in the control groups. In addition, strawberry extracts were evaluated for their antiproliferative activities using HT-29 colon cancer cells. Extracts from all treated fruit exhibited significantly stronger inhibition on HT-29 cell proliferation than those from the control fruit. These data provide evidence that, in addition to possessing antimicrobial activity, the essential oils also increase free radical scavenging capacity and antiproliferative activity in fruit and, in turn, enhance the resistance of fruit tissues to deterioration and spoilage.

Inhibitory effect on activator protein-1, nuclear factor-kappaB, and cell transformation by extracts of strawberries (Fragaria x ananassa Duch.).

The inhibitory effects of strawberry (Fragaria x ananassa Duch.) antioxidant enzymes on tetradecanoylphorbol-13-acetate (TPA) or ultraviolet-B (UVB) induced activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) were studied. The inhibitory effects of strawberry extracts on the proliferation and transformation of human and mouse cancer cells were also evaluated. Strawberries had high activities of glutathione peroxidase, superoxide dismutase, guaiacol peroxidase, ascorbate peroxidase, and glutathione reductase. Strawberry extracts inhibited the proliferation of human lung epithelial cancer cell line A549 and decreased TPA-induced neoplastic transformation of JB6 P+ mouse epidermal cells. Pretreatment of JB6 P+ mouse epidermal cells with strawberry extract resulted in the inhibition of both UVB- and TPA-induced AP-1 and NF-kappaB transactivation. Furthermore, strawberry extract also blocked TPA-induced phosphorylation of extracellular signal-regulated kinases (ERKs) and UVB-induced phosphorylation of ERKs and JNK kinase in JB6 P+ mouse epidermal cell culture. These results suggest that the ability of strawberries to block UVB- and TPA-induced AP-1 and NF-kappaB activation may be due to their antioxidant properties and their ability to reduce oxidative stress. The oxidative events that regulate AP-1 and NF-kappaB transactivation can be important molecular targets for cancer prevention. The strawberries may be highly effective as a chemopreventive agent that acts by targeting the down-regulation of AP-1 and NF-kappaB activities, blocking MAPK signaling, and suppressing cancer cell proliferation and transformation.

Antioxidant activity in lingonberries (Vaccinium vitis-idaea L.) and its inhibitory effect on activator protein-1, nuclear factor-kappaB, and mitogen-activated protein kinases activation.

Lingonberry has been shown to contain high antioxidant activity. Fruits from different cultivars of lingonberry (Vaccinium vitis-idaea L.) were evaluated for fruit quality, antioxidant activity, and anthocyanin and phenolic contents. The fruit soluble solids, titratable acids, antioxidant capacity, and anthocyanin and phenolic contents varied with cultivars. Lingonberries contain potent free radical scavenging activities for DPPH*, ROO*, *OH, and O2*- radicals. Pretreatment of JB6 P+ mouse epidermal cells with lingonberry extracts produced a dose-dependent inhibition on the activation of activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) induced by either 12-O-tetradecanoylphorbol-13-acetate (TPA) or ultraviolet-B (UVB). Lingonberry extract blocked UVB-induced phosphorylation of the mitogen-activated protein kinase (MAPK) signaling members ERK1, ERK2, p38, and MEK1/2 but not JNK. Lingonberry extract also prevented TPA-induced phosphorylation of ERK1, ERK2, and MEK1/2. Results of soft agar assays indicated that lingonberry extract suppressed TPA-induced neoplastic transformation of JB6 P(+) cells in a dose-dependent manner. Lingonberry extract also induced the apoptosis of human leukemia HL-60 cells in a dose-independent manner. These results suggest that ERK1, ERK2, and MEK1/2 may be the primary targets of lingonberry that result in suppression of AP-1, NF-kappaB, and neoplastic transformation in JB6 P(+) cells and causes cancer cell death by an apoptotic mechanism in human leukemia HL-60 cells.

Oxygen radical absorbing capacity of phenolics in blueberries, cranberries, chokeberries, and lingonberries.

The antioxidant activity of phenolics in fruits of blueberry (Vaccinium corymbosum cv. Sierra), cranberry (Vaccinium macrocarpon cv. Ben Lear), wild chokeberry (Aronia melanocarpa), and lingonberry (Vaccinium vitis-idaea cv. Amberland) was determined in this study. The phenolic constituents and contents among the different berries varied considerably. Anthocyanins were found to be the main components in all these berries. Chlorogenic acid in blueberry, quercetin glycosides in cranberry and lingonberry, and caffeic acid and its derivative in chokeberry were also present in relatively high concentrations. Chlorogenic acid, peonidin 3-galactoside, cyanidin 3-galactoside, and cyanidin 3-galactoside were the most important antioxidants in blueberry, cranberry, wild chokeberry, and lingonberry, respectively. The contribution of individual phenolics to the total antioxidant capacity was generally dependent on their structure and content in the berries. Phenolics such as quercetin and cyanidin, with 3',4'-dihydroxy substituents in the B ring and conjugation between the A and B rings, had highly effective radical scavenging structures in blueberries, cranberries, chokeberries, and lingonberries. Phenolic acids such as caffeic acid also showed high antioxidant activity, probably due to its dihydroxylation in the 3,4 positions as hydrogen donors.

Compost as a soil supplement increases the level of antioxidant compounds and oxygen radical absorbance capacity in strawberries.

Compost as a soil supplement significantly enhanced levels of ascorbic acid (AsA) and glutathione (GSH) and ratios of AsA/dehydroascorbic acid (DHAsA) and GSH/oxidized glutathione (GSSG) in fruit of two strawberry (Fragaria x ananassa Duch.) cultivars, Allstar and Honeoye. The peroxyl radical (ROO(*)) as well as the superoxide radical (O(2)(*)(-)), hydrogen peroxide (H(2)O(2)), hydroxyl radical (OH(*)), and singlet oxygen ((1)O(2)) absorbance capacity in strawberries increased significantly with increasing fertilizer strength and compost use. The planting medium (compost) x fertilizer interaction for phenolics and flavonoids was significant. Fruit from plants grown in full-strength fertilizer with 50% soil plus 50% compost and 100% compost yielded fruit with the highest levels of phenolics, flavonol, and anthocyanin content. A positive relationship between antioxidant activities and contents of AsA and GSH and ratios of AsA/DHAsA and GSH/GSSG existed in fruit of both strawberry cultivars. Correlation coefficients for the content of antioxidant components versus antioxidant activity [against ROO(*), O(2)(*)(-), H(2)O(2), OH(*), or (1)O(2)] ranged from r( )()= 0.7706 for H(2)O(2) versus GSH/GSSH in cv. Allstar to r = 0.9832 for O(2)(*)(-) versus total flavonoids in cv. Allstar.

Cultural system affects fruit quality and antioxidant capacity in strawberries.

Cultural system [hill plasticulture (HC) versus matted row (MR)] and genotype interactions affected strawberry fruit quality. In general, fruit soluble solids content, total sugar, fructose, glucose, ascorbic acid, titratable acid, and citric acid contents were increased in the HC system. Fruit from HC also had higher flavonoid contents and antioxidant capacities. Strawberry fruit contains flavonols as well as other phenolic compounds such as anthocyanins and phenolic acids. Pelargonidin-based anthocyanins such as pelargonidin 3-glucoside, pelargonidin 3-rutinoside, and pelargonidin 3-glucoside-succinate were the predominant anthocyanins in strawberry fruit. The content of cyanidin-based anthocyanins, cyanidin 3-glucoside and cyanidin 3-glucoside-succinate, was much lower than that of pelargonidin-based anthocyanins in either system. Strawberry fruit from the HC system had significantly higher amounts of p-coumaroylglucose, dihydroflavonol, quercetin 3-glucoside, quercetin 3-glucuronide, kaempferol 3-glucoside, kaempferol 3-glucuronide, cyanidin 3-glucoside, pelargonidin 3-glucoside, pelargonidin 3-rutinoside, cyanidin 3-glucoside-succinate, and pelargonidin 3-glucoside-succinate. Fruits from plants grown in the MR system generally had the lowest contents of phenolic acids, flavonols, and anthocyanins. Strawberry fruit grown under HC conditions had significantly higher peroxyl radicals (ROO*) absorbance capacity (ORAC).

Elevated carbon dioxide increases contents of antioxidant compounds in field-grown strawberries.

The effects of elevated CO2 concentrations on the antioxidant capacity and flavonoid content in strawberry fruit (Fragaria x ananassa Duch.) were studied under field conditions. Increased CO(2) (300 and 600 micromol mol(-1) above ambient) concentrations resulted in increases in ascorbic acid (AsA), glutathione (GSH), and ratios of AsA to dehydroascorbic acid (DHAsA) and GSH to oxidized glutathione (GSSG), and a decrease in DHAsA in strawberry fruit. High anthocyanin and phenolic content were also found in fruit of CO(2) treated plants. Growing strawberry plants under CO(2) enrichment conditions significantly enhanced fruit p-coumaroylglucose, dihydroflavonol, quercetin 3-glucoside, quercetin 3-glucuronide, and kaempferol 3-glucoside contents, as well as cyanidin 3-glucoside, pelargonidin 3-glucoside, and pelargonidin 3-glucoside-succinate content. Fruit of strawberry plants grown in the CO(2) enrichment conditions also had high oxygen radical absorbance activity against ROO(*), O(2)(*-), H(2)O(2), OH(*), and (1)O(2) radicals.

Effect of high-oxygen atmospheres on blueberry phenolics, anthocyanins, and antioxidant capacity.

The influence of high oxygen concentrations on total phenolic, total anthocyanin, individual phenolic compounds, and antioxidant capacity (measured as oxygen radical absorbance capacity, ORAC) in highbush blueberry fruit (Vaccinium corymbosum L. cv. Duke) was investigated. Freshly harvested blueberries were placed in jars ventilated continuously with air or with 40, 60, 80, or 100% O(2) at 5 degrees C for up to 35 days. Samples were taken initially and at 7-day intervals during storage. Whereas the quality parameters of titratable acidity, total soluble solids, and surface color were only slightly affected by the superatmospheric O(2) treatments, the antioxidant levels were markedly increased by 60-100% O(2) treatments as compared with 40% O(2) treatment or air control during 35 days of storage. Elevated O(2) between 60 and 100% also promoted increases of total phenolics and total anthocyanins as well as the individual phenolic compounds analyzed by HPLC. Fruit treated with O(2) concentrations of >/=60% also exhibited significantly less decay. Data obtained in this study suggest that high-oxygen treatments may improve the antioxidant capacity of blueberry fruit. Furthermore, antioxidant capacity may be correlated with total phenolic and anthocyanin contents in blueberries.

Effect of cultural system and essential oil treatment on antioxidant capacity in raspberries.

The effects of cultural system and essential oil treatment on antioxidant capacities in raspberries were evaluated. Raspberries were hand-harvested from organic and conventional farms in Maryland, USA, and were treated with essential oil including carvacrol, anethole, cinnamic acid, perillaldehyde, cinnamaldehyde, and linalool. Results from this study showed that raspberries grown from organic culture exhibited higher value of antioxidant capacities and individual flavonoids contents. Moreover, the organic culture also enhanced the activities of antioxidant enzymes. In addition, essential oil treatments promoted the antioxidant enzymes activities and antioxidant capacities of raspberries, and the most effective compound was perillaldehyde. In conclusion, raspberries produced from organic culture contained significantly higher antioxidant capacities than those produce from conventional culture. Postharvest essential oil treatments have positive effect on enhancing antioxidant capacities in raspberries from both organic and conventional cultures.

Delphinidin induces necrosis in hepatocellular carcinoma cells in the presence of 3-methyladenine, an autophagy inhibitor.

The present study was performed to determine whether anthocyanins could trigger different modes of cell death in different cancers. It was found that whereas cyanidin-3-rutinoside and delphinidin could induce apoptosis in leukemia cells, they caused growth retardation in hepatocellular carcinoma cells (HCC), which was accompanied with a significant cellular vacuolization. The latter was likely caused by macroautophagy and was completely suppressed by 3-methyladenine, an inhibitor of class III phosphoinositide 3-kinase that is important for autophagy activation, and by bafilomycin A1, which blocks lysosomal degradation. Delphinidin induced significant lipidation of LC3, an indication of macroautophagy, which was also suppressed by 3-methyladenine. Macroautophagy was required for the survival of delphinidin-treated HCC cells as inhibition with 3-methyladenine led to massive necrosis without caspase activation. Thus, anthocyanins could induce different modes of cell death for different cancers. Furthermore, anthocyanins could be used in combination with a macroautophagy inhibitor for treating cancers such as HCC.

Effect of different cultural systems on antioxidant capacity, phenolic content, and fruit quality of strawberries (Fragaria x aranassa Duch.).

The effect of cultivation practices for controlling strawberry black root rot (BRR) on fruit quality, antioxidant capacity, and flavonoid content in two strawberry cultivars Allstar and Chandler (Fragaria x ananassa Duch.) was evaluated. Strawberry fruits used in this study were from plants grown in soils which had a prior history of BRR and red stele, and had not been fumigated during the seven years prior to the study. Results from this study showed that fruit from plants grown in compost socks had significantly higher oxygen radical absorbance capacity (ORAC), flavonoids, anthocyanins, soluble solids content (SSC), titratable acid (TA), fructose, glucose, sucrose, malic acid, and citric acid than fruit produced in the black plastic mulch or matted row systems. Cultivar Chandler surpassed cv. Allstar in sugar content, acid content, and flavonoid content regardless of preplanting vinegar drenching and various culture treatments. However, preplanting vinegar treatment increased cyanidin-based and pelargonidin-based anthocyanins but decreased sugar content in fruits of both cultivars.

Fruit quality, antioxidant capacity, and flavonoid content of organically and conventionally grown blueberries.

The effect of cultivation practices on fruit quality and antioxidant capacity in highbush blueberries var. Bluecrop (Vaccinium corymbosum L.) was evaluated from random samples of commercial late harvest fields in New Jersey. Results from this study showed that blueberry fruit grown from organic culture yielded significantly higher sugars (fructose and glucose), malic acid, total phenolics, total anthocyanins, and antioxidant activity (ORAC) than fruit from the conventional culture. In organically cultured fruit, the average values for the ORAC, total anthocyanins, and total phenolic content were 46.14 micromol of Trolox (TE)/g of fresh weight (fwt), 131.2 mg/100 g of fwt, and 319.3 mg/100 g of fwt, respectively. In conventionally cultured fruit, the average values for the ORAC, total anthocyanin, and total phenol content were 30.8 micromol of TE/g of fwt, 82.4 mg/100 g of fwt, and 190.3 mg/100 g of fwt, respectively. The organic culture also produced fruit with higher contents of myricetin 3-arabinoside, quercetin 3-glucoside, delphinidin 3-galactoside, delphinidin 3-glucoside, delphinidin 3-arabinoside, petunidin 3-galactoside, petunidin 3-glucoside, and malvidin 3-arabinoside than conventional culture. There was a significant correlation between the ORAC values and total phenolics and total anthocyanins. These results indicate that even though there were variations in phytonutrient content among individual farms within each cultural system, significant differences between two cultivation practices were evident.

Variations in free radical scavenging capacity and antiproliferative activity among different genotypes of autumn olive (Elaeagnus umbellata).

Fruit from six genotypes of autumn olive (Elaeagnus umbellata Thunb.) ('Brilliant Rose', 'Delightful', 'Jewel', Natural 1, Natural 2, and 'Sweet Tart') were evaluated for antioxidant capacity and anti-cancer properties. Based on data from electron spin resonance (ESR) measurements, autumn olive contained potent free radical scavenging activities for hydroxyl (*OH) and superoxide (O2*-) radicals. Among the six genotypes, 'Brilliant Rose' and 'Jewel' had the highest levels of antioxidant activity. Pretreatment of JG6 P+ mouse epidermal cells with autumn olive extracts inhibited the activation of activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) induced by either 12-O-tetradecanoylphorbol 13-acetate (TPA) or ultraviolet-B (UVB). Extracts of all autumn olive genotypes inhibited proliferation of human leukemia HL-60 cancer cells and human lung epithelial cancer A549 cells and induced apoptosis of HL-60 cells. In particular, 'Brilliant Rose' and 'Jewel' had relatively potent activities compared to other genotypes. These results indicate that consuming autumn olive fruit may be beneficial to human health, although further studies are needed for confirmation.

Antioxidant activity of Vaccinium stamineum: exhibition of anticancer capability in human lung and leukemia cells.

Fruit of deerberry [Vaccinium stamineum L.] were evaluated for their antioxidant capacity and anticancer properties in JB6 P (+) mouse epidermal cells, human lung and leukemia cells. Deerberries contain potent free radical scavenging activities. Pretreatment of JB6 P (+) mouse epidermal cells with deerberry fruit extracts produced an inhibition on the activation of activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) induced by either 12- O-tetradecanoylphorbol 13-acetate (TPA) or ultraviolet-B (UVB). Deerberry fruit extracts also blocked TPA- or UVB-induced phosphorylation of ERKs and MEK 1/2, two upstream regulators of AP-1 and inhibited proliferation of human leukemia HL-60 cancer cells and human lung epithelial cancer A549 cells and induced apoptosis of HL-60 cells. These results suggest that the inhibition of TPA- or UVB-induced AP-1 and NF-kappaB activity, inhibition of HL-60 cells and cancer A549 cells proliferation and induction of apoptotic in human leukemia HL-60 cancer cells may be mediated through the ERKs and MEK 1/2 signal pathway.

Cyanidin-3-rutinoside, a natural polyphenol antioxidant, selectively kills leukemic cells by induction of oxidative stress.

Anthocyanins are a group of naturally occurring phenolic compounds widely available in fruits and vegetables in human diets. They have broad biological activities including anti-mutagenesis and anticarcinogenesis, which are generally attributed to their antioxidant activities. We studied the effects and the mechanisms of the most common type of anthocyanins, cyanidin-3-rutinoside, in several leukemia and lymphoma cell lines. We found that cyanidin-3-rutinoside extracted and purified from the black raspberry cultivar Jewel induced apoptosis in HL-60 cells in a dose- and time-dependent manner. Paradoxically, this compound induced the accumulation of peroxides, which are involved in the induction of apoptosis in HL-60 cells. In addition, cyanidin-3-rutinoside treatment resulted in reactive oxygen species (ROS)-dependent activation of p38 MAPK and JNK, which contributed to cell death by activating the mitochondrial pathway mediated by Bim. Down-regulation of Bim or overexpression of Bcl-2 or Bcl-x(L) considerably blocked apoptosis. Notably, cyanidin-3-rutinoside treatment did not lead to increased ROS accumulation in normal human peripheral blood mononuclear cells and had no cytotoxic effects on these cells. These results indicate that cyanidin-3-rutinoside has the potential to be used in leukemia therapy with the advantages of being widely available and selective against tumors.

Cyanidin-3-glucoside, a natural product derived from blackberry, exhibits chemopreventive and chemotherapeutic activity.

Epidemiological data suggest that consumption of fruits and vegetables has been associated with a lower incidence of cancer. Cyanidin-3-glucoside (C3G), a compound found in blackberry and other food products, was shown to possess chemopreventive and chemotherapeutic activity in the present study. In cultured JB6 cells, C3G was able to scavenge ultraviolet B-induced *OH and O2-* radicals. In vivo studies indicated that C3G treatment decreased the number of non-malignant and malignant skin tumors per mouse induced by 12-O-tetradecanolyphorbol-13-acetate (TPA) in 7,12-dimethylbenz[a]anthracene-initiated mouse skin. Pretreatment of JB6 cells with C3G inhibited UVB- and TPA-induced transactivation of NF-kappaB and AP-1 and expression of cyclooxygenase-2 and tumor necrosis factor-alpha. These inhibitory effects appear to be mediated through the inhibition of MAPK activity. C3G also blocked TPA-induced neoplastic transformation in JB6 cells. In addition, C3G inhibited proliferation of a human lung carcinoma cell line, A549. Animal studies showed that C3G reduced the size of A549 tumor xenograft growth and significantly inhibited metastasis in nude mice. Mechanistic studies indicated that C3G inhibited migration and invasion of A549 tumor cells. These finding demonstrate for the first time that a purified compound of anthocyanin inhibits tumor promoter-induced carcinogenesis and tumor metastasis in vivo.