Hypoglycemic effects of a phenolic acid fraction of rice bran and ferulic acid in C57BL/KsJ-db/db mice

Rice bran contains many phenolic acids, the most abundant of which is the antioxidant, ferulic acid (FA). We evaluated the hypoglycemic effects of a phenolic acid fraction (the ethyl acetate fraction, EAE) of rice bran and of FA in C57BL/KsJ db/db mice. Type 2 diabetic mice were allocated to a control group, an EAE group, or an FA group. Animals were fed a modified AIN-76 diet, and EAE or FA was administered orally for 17 days. There was no significant difference in body weight gain between groups. Administration of EAE and FA significantly decreased blood glucose levels and increased plasma insulin levels. EAE or FA groups had significantly elevated hepatic glycogen synthesis and glucokinase activity compared with the control group. Plasma total cholesterol and low density lipoprotein (LDL) cholesterol concentrations were significantly decreased by EAE and FA administration. These findings suggest that EAE and FA may be beneficial for treatment of type 2 diabetes because they regulate blood glucose levels by elevating glucokinase activity and production of glycogen in the liver.     

Hydrogen peroxide is required for poly(phenolic) domain formation during wound-induced suberization

The requirement for hydrogen peroxide (H(2)O(2)) during suberization was demonstrated in wound-induced potato tubers by monitoring the extent of phenolic polymerization after the inhibition of H(2)O(2) production using diphenyleneiodonium (DPI). In DPI-treated tissues the extent of phenolic polymerization in suberized tissues, measured using DFRC (Derivatization Followed by Reductive Cleavage) and thioglycolic acid analyses, was greatly reduced relative to untreated controls. Concomitantly, a large quantity of new soluble phenolics accumulated in the DPI-treated tissue some of which were not present in the controls. We suggest that the inhibition of H(2)O(2) production prevented these phenolics from being oxidized by cell wall peroxidases. As a result, these phenolics were left unpolymerized and accumulated in the tissue. Several of the soluble phenolics were identified as hydroxycinnamic acid derivatives. From the data presented, it was concluded that H(2)O(2) is required for the polymerization step in the formation of the poly(phenolic) domain of suberized potato tubers.     

Antioxidant and antiulcerative properties of phenolics from Chinese quince, quince, and apple fruits

To evaluate the health benefits of Chinese quince and quince phenolics, their antioxidant properties and antiulcerative activity were investigated in comparison with apple phenolics as a reference. The strength of antioxidant activity and DPPH radical scavenging activity of these fruit phenolics varied according to different in vitro evaluation systems, whereas the antioxidative property of rat blood increased in all rats orally administered phenolics. Ferulic acid and isoferulic acid were detected as major metabolites in rats given apple phenolics, quince phenolics, and 5-caffeoylquinic acid standard. (-)-Epicatechin and its 3'-O-methyl ether could be detected in rats administered apple phenolics and (-)-epicatechin standard. In the ethanol-induced gastric ulcer, pre-administration of Chinese quince and quince phenolics suppressed the occurrence of gastric lesions in rats, whereas apple phenolics seemed to promote ulceration. The trend of myeloperoxidase activity was similar to that of the ulcer index. The results showed that Chinese quince and quince phenolics might have health benefits by acting both in blood vessels and on the gastrointestinal tract.     

Accurate assessment of the bioactivities of redox-active polyphenolics in cell culture

Phenolic compounds are widely known for their roles as antioxidants and anti-inflammatory agents, as well as for their epidemiological association with reduced risks for certain types of diseases. In the present study, we used rabbit peripheral blood mononuclear cells (PBMCs) to evaluate possible artifacts that result from the reactivity of polyphenolics. We evaluated several common methods for cytotoxicity tests using nine polyphenolics, representing several major classes of tannins and their subunits. For three of those phenolics, we investigated whether or not the bioactivities of the phenolics were altered by spontaneous oxidation. Our study showed that many of the nine tested tannins interfered with the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay, which is commonly used to measure cell viability. A better method for determining cell viability is the luciferin/luciferase ATP assay, and using that method, we found that several tannins are toxic to PBMCs. We measured TNF-alpha production to assess possible anti-inflammatory activity, and found that only apigenin inhibited TNF-alpha production in LPS-stimulated cells (EC 50 1.0 microg/mL). The other polyphenolic compounds we tested either had no effect on TNF-alpha or increased its production. However, our data indicated that spontaneous oxidation altered the activity of phenolics, eliminating their toxicity. This study shows that the chemical reactivity of phenolics can significantly affect attempts to evaluate bioactivity in cultured cells and that particular attention should be paid to both methods for determining toxicity and to spontaneous oxidation of tannins during cell testing.     

Antioxidant activity of minimally processed red chicory (Cichorium intybus L.) evaluated in xanthine oxidase-, myeloperoxidase-, and diaphorase-catalyzed reactions

Minimally processed red chicory products (Cichorium intybus L. var. silvestre) were studied for their polyphenol content and antioxidant activity evaluated by using the synthetic 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl radical and three model reactions catalyzed by relevant enzymatic sources of reactive oxygen species, namely, xanthine oxidase, myeloperoxidase, and diaphorase. Products were analyzed at the time of production and after storage at 4 degrees C within either a gas permeable film or a gas barrier film. The antioxidant activity and contents of hydroxycinnamic acids and flavonoids decreased by less than 20% during storage of the minimally processed red chicory products. Total phenolics were significantly correlated with the antioxidant activity evaluated with both the synthetic radical and the enzyme-catalyzed reactions. On a molar basis, red chicory phenolics were as efficient as the reference compound Trolox in scavenging the synthetic radical. However, red chicory phenolics had a much higher inhibitory activity than Trolox in the model enzymatic systems.     

Blanching Improves Anthocyanin Absorption from Highbush Blueberry ( Vaccinium corymbosum L.) Purée in Healthy Human Volunteers: A Pilot Study

Blueberries ( Vaccinium corymbosum L.) are rich sources of phenolics and anthocyanins (ACNs). We investigated the absorption of ACNs after consumption of one portion (300 g) of minimally processed blueberry purée (P) obtained from blanched (BL) or unblanched (NB) berries. A repeated-measures, crossover design study was conducted on healthy human volunteers. Blood was drawn between baseline and 24 h after BL-P or NB-P consumption, while urine were collected from the day before the experiment up to 48 h. Total plasma ACN content was not significantly different, while phenolics content was higher in BL-P with respect to NB-P. The maximum ACN absorption in plasma was observed after 1.5 h from the intake of the purées and was significantly higher (p ≤ 0.05) after the intake of BL-P. Both products increased the excretion of hippuric acid in urine. In conclusion, blanching had no significant effect on total ACN content and enhanced their absorption from minimally processed purées.     

Antioxidant capacity in cranberry is influenced by cultivar and storage temperature

Ten cranberry (Vaccinium macrocarpon Aiton) cultivars were evaluated for oxygen radical absorbance capacity (ORAC), anthocyanins, and total phenolics contents after three months of storage at 0, 5, 10, 15, and 20 degrees C. The antioxidant capacity of cranberry was affected by cultivars and storage temperatures. Among the 10 cranberry cultivars used in this study, Early Black, Crowley, and Franklin had higher antioxidant capacities than the other cultivars. ORAC values, anthocyanins, and total phenolics contents increased during storage. The highest increases in antioxidant activity, anthocyanin, and phenolics contents occurred at 15 degrees C storage. Fruit stored at 20 degrees C had lower ORAC values than those stored at 15 degrees C. A positive relationship existed between ORAC values and anthocyanin or phenolic content in all 10 cranberry cultivars at different storage temperatures.     

Enzymatic production of galactooligosaccharides by beta-galactosidase from Bifidobacterium longum BCRC 15708

The production of galactooligosaccharides (GOSs) by transgalactosylation using beta-galactosidase from Bifidobacterium longum BCRC 15708 was studied. Other than lactose, galactose, and glucose, two types of GOSs, tri- and tetrasaccharides, were formed after beta-galactosidase action on 40% lactose. Trisaccharides were the major type of GOS formed. Generally, an increase of the initial lactose concentration in the reaction mixture resulted in a higher GOS production. A maximum yield of 32.5% (w/w) GOSs could be achieved from 40% lactose solution at 45 degrees C, pH 6.8, when the lactose conversion was 59.4%. The corresponding productivity of GOSs was 13.0 g/(L.h). Transgalactosylation activity of beta-galactosidase from a test organism showed a relatively lower sensitivity toward glucose and galactose than that from other organisms. The addition of 5% or 10% glucose or galactose to the reaction mixture did not significantly (p>0.05) reduce the transgalactosylation reaction of beta-galactosidase.     

Simple and rapid method for the analysis of phenolic compounds in beverages and grains

A new method for the detection of phenolics in food systems was developed. This method is based on interactions of phenolics with Fast Blue BB diazonium salt in alkali pH, forming azo complexes, with the absorbance measured at 420 nm after 60 min. The linear regression correlations (R(2)) of gallic acid calibration standards were >0.99. The phenolic content (gallic acid equivalent) of samples analyzed yielded higher ratios (1.7-6.6) of the total phenolics by Fast Blue BB to Folin-Ciocalteu methods in most beverages and grain samples, but in flaxseed and some juice blends, the ratios were <1. The lower ratios suggest the presence of non-phenolic reducing constituents measured with the Folin-Ciocalteu method as "total phenolics". This method is simple and inexpensive and can be used to rapidly assess the total phenolics of foods and beverages.     

Impact of suppression of ethylene action or biosynthesis on flavor metabolites in apple (Malus domestica Borkh) fruits

To understand the role of ethylene in regulating the overall flavor of apple fruits, ethylene production or action was reduced using transgenic apple trees suppressed for ACC-synthase or ACC-oxidase enzyme activity or by the addition of 1-methylcyclopropene (1-MCP), an ethylene action inhibitor. Flavor components were differentially regulated in response to the suppression of both ethylene biosynthesis and action. Headspace analysis of aroma production, an ethylene-associated event, showed a reduction in ester and alcohol production in the ethylene-suppressed lines and in the apples treated with 1 microL L(-1) 1-MCP for 20 h at 20 degrees C. However, no major differences were observed in the concentrations of aldehyde volatiles. Other flavor metabolites that showed an ethylene-dependent pattern were organic acids and sugars. Malic acid degradation was significantly reduced under ethylene-suppressed conditions, showing a recovery after the fruit was exposed to ethylene. Sucrose and fructose concentrations were influenced by suppression or enhancement of ethylene. Total phenolics as well as individual phenolic compounds showed an ethylene-dependent regulation only in response to the suppression of ethylene biosynthesis, but not when ethylene action was inhibited.     

Processing strawberries to different products alters contents of vitamin C, total phenolics, total anthocyanins, and antioxidant capacity

Strawberries were processed to juice, nectar, wine, and puree. For investigation of the antioxidant capacity as well as the contents of ascorbic acid, total phenolics and total anthocyanins, samples were taken after different stages of production to determine the effects of processing. The content of vitamin C was measured spectrophotometrically. The total phenolic content was analyzed by using the Folin-Ciocalteu method, and the amount of total anthocyanins was determined by using the pH-differential method. Two different methods-the trolox equivalent antioxidant capacity assay and the ferric reducing antioxidant power test-were used to determine the hydrophilic antioxidant capacity. This study showed the decrease of all investigated parameters within processing strawberries to different products. The content of ascorbic acid decreased with production time and processing steps, especially during heat treatment. The investigations on total phenolics in strawberry products proved fining to be a mild method to clarify berry juices and wines without removing high amounts of total phenolics. Fermentation did not lead to heavy losses of total phenolics, probably due to polymerization and condensation of monomer phenolics such as anthocyanins. Total anthocyanins and the hydrophilic antioxidant capacity decreased while using high temperatures. Anthocyanins also decreased considerably during the processing of wines, mainly caused by fermentation and pasteurization.     

Determination of phenolic compounds and ascorbic acid in different fractions of tomato by capillary electrophoresis with electrochemical detection

Tomato (Lycopersicon esculentum Mill.), one of the most important crops worldwide, contains different classes of substances with antioxidant properties such as carotenoids, vitamin C, and phenolics. A method based on capillary electrophoresis with electrochemical detection has been developed to analyze ascorbic acid and phenolics in the peel, pulp, and seeds of tomatoes. Operating in a wall-jet configuration, a 300 microm diameter carbon disk electrode was used as the working electrode, which exhibits a good response at +0.90 V (vs saturated calomel electrode) for the analytes. Under optimum conditions, the analytes were baseline separated within 20 min in a 50 mmol/L borate buffer (pH 8.7). Notably, excellent linearity was obtained over 3 orders of magnitude with detection limits (S/N=3) ranging from 1x10(-8) to 2x10(-7) g/mL for all analytes. This proposed method has been successfully applied to monitor the content of ascorbic acid and phenolics in real samples, and the assay results were satisfactory.     

Purple carrot (Daucus carota L.) polyacetylenes decrease lipopolysaccharide-induced expression of inflammatory proteins in macrophage and endothelial cells

Carrots ( Daucus carota L.) contain phytochemicals including carotenoids, phenolics, polyacetylenes, isocoumarins, and sesquiterpenes. Purple carrots also contain anthocyanins. The anti-inflammatory activity of extracts and phytochemicals from purple carrots was investigated by determining attenuation of the response to lipopolysaccharide (LPS). A bioactive chromatographic fraction (Sephadex LH-20) reduced LPS inflammatory response. There was a dose-dependent reduction in nitric oxide production and mRNA of pro-inflammatory cytokines (IL-6, IL-1beta, TNF-alpha) and iNOS in macrophage cells. Protein secretions of IL-6 and TNF-alpha were reduced 77 and 66% in porcine aortic endothelial cells treated with 6.6 and 13.3 microg/mL of the LH-20 fraction, respectively. Preparative liquid chromatography resulted in a bioactive subfraction enriched in the polyacetylene compounds falcarindiol, falcarindiol 3-acetate, and falcarinol. The polyacetylenes were isolated and reduced nitric oxide production in macrophage cells by as much as 65% without cytotoxicity. These results suggest that polyacetylenes, not anthocyanins, in purple carrots are responsible for anti-inflammatory bioactivity.     

Reddish coloration of Chinese quince (Pseudocydonia sinensis) procyanidins during heat treatment and effect on antioxidant and antiinfluenza viral activities

To investigate the effects of processing Chinese quince fruit on the denaturation of phenolics and their food functions, fruit phenolic extracts were heated together with organic acid for up to 12 h. Chinese quince phenolic (mostly procyanidins) solution subjected to heat treatment changed from almost colorless, pale yellow, to a reddish color. Before heat treatment, the absorption spectra of polymeric procyanidins were observed only around 280 nm; after heat treatment, absorption occurred between 400 and 600 nm, which is related to the reddish color appearance. Thioacidolysis of denatured reddish phenolics showed that (-)-epicatechin subunits decreased during heat treatment and, in contrast, cyanidin increased. In addition, novel substances that could not be degraded by thioacidolysis were formed. Meanwhile, antioxidant activities, assessed by linoleic acid peroxidation, 1,1-diphenyl-2-picrylhydrazyl (DPPH), Folin-Ciocalteu, and FRAP methods, increased during heat treatment. The antiinfluenza viral activity of denatured reddish phenolics was inferior to that of intact fruit phenolics; however, they retained moderate activity. These results indicate that red coloration of fruit products of Chinese quince was mainly due to the spectral (i.e. structural) changes of procyanidins accompanied with formation of cyanidin. Increasing the length of heat treatment increased the antioxidant capacity of phenolics, and the resultant reddish phenolics retained moderate antiinfluenza viral activity.     

Isolation and identification of strawberry phenolics with antioxidant and human cancer cell antiproliferative properties

Studies suggest that consumption of berry fruits, including strawberries ( Fragaria x ananassa Duch.), may have beneficial effects against oxidative stress mediated diseases such as cancer. Berries contain multiple phenolic compounds, which are thought to contribute to their biological properties. Comprehensive profiling of phenolics from strawberries was previously reported using high-performance liquid chromatography with mass spectrometry (HPLC-MS) detection. The current study reports the isolation and structural characterization of 10 phenolic compounds from strawberry extracts using a combination of Amberlite XAD16-resin and C18 columns, HPLC-UV, and nuclear magnetic resonance (NMR) spectroscopy methods. The phenolics were cyanidin-3-glucoside ( 1), pelargonidin (2), pelargonidin-3-glucoside (3), pelargonidin-3-rutinoside (4), kaempferol (5), quercetin (6), kaempferol-3-(6'-coumaroyl)glucoside) (7), 3,4,5-trihydroxyphenyl-acrylic acid (8), glucose ester of ( E)- p-coumaric acid (9), and ellagic acid . Strawberry crude extracts and purified compounds 1- 10 were evaluated for antioxidant and human cancer cell antiproliferative activities by the Trolox equivalent antioxidant capacity (TEAC) and luminescent ATP cell viability assays, respectively. Among the pure compounds, the anthocyanins 1 (7156 microM Trolox/mg), 2 (4922 microM Trolox/mg), and 4 (5514 microM Trolox/mg) were the most potent antioxidants. Crude extracts (250 microg/mL) and pure compounds (100 microg/mL) inhibited the growth of human oral (CAL-27, KB), colon (HT29, HCT-116), and prostate (LNCaP, DU145) cancer cells with different sensitivities observed between cell lines. This study adds to the growing body of data supporting the bioactivities of berry fruit phenolics and their potential impact on human health.     

Insulin-releasing properties of a series of cinnamic acid derivatives in vitro and in vivo

Cinnamic acid derivatives are naturally occurring substances found in fruits, vegetables, and flowers and are consumed as dietary phenolic compounds. In the present study, cinnamic acid and its derivatives were evaluated for insulin secreting activity in perfused rat pancreas and pancreatic beta-cells (INS-1) as well as an increase in [Ca(2+)]i in vitro. The presence of m-hydroxy or p-methoxy residues on cinnamic acid was a significantly important substituent as an effective insulin releasing agent. The introduction of p-hydroxy and m-methoxy-substituted groups in cinnamic acid structure (ferulic acid) displayed the most potent insulin secreting agent among those of cinnamic acid derivatives. In particular, the stimulatory insulin secreting activities of test compounds were associated with a rise of [Ca(2+)]i in INS-1. In perfused rat pancreas, m-hydroxycinnamic acid, p-methoxycinnamic acid, and ferulic acid (100 microM) significantly stimulated insulin secretion during 10 min of administration. The onset time of insulin secretion of those compounds was less than 1 min and reached its peak at 4 min that was about 2.8-, 3.3-, and 3.4-fold of the baseline level, respectively. Intravenous administration of p-methoxycinnamic acid and ferulic acid (5 mg/kg) significantly decreased plasma glucose and increased insulin concentration in normal rats and maintained its level for 15 min until the end of experiment. Meanwhile, m-hydroxycinnamic acid induced a significant lowering of plasma glucose after 6 min, but the effects were transient with plasma glucose concentration, rapidly returning to basal levels. Our findings suggested that p-methoxycinnamic acid and ferulic acid may be beneficial for the treatment of diabetes mellitus because they regulated blood glucose level by stimulating insulin secretion from pancreatic beta-cells.     

DHA对脂肪细胞周期及COX-2 mRNA表达的调节

以大鼠前体脂肪细胞原代单层培养为模型，分别以0(对照组)、40(低剂量组)和160 μmol/L(高剂量组)的二十二碳六烯酸(DHA)处理未分化的前体脂肪细胞(培养第1天)和分化的脂肪细胞(分化第1天)。采用流式细胞仪(FCM)检测细胞周期分布；逆转录-聚合酶链反应(RT-PCR)分析环氧合酶-2(COX-2 )mRNA表达情况，探讨DHA对脂肪细胞周期及基因表达的调节作用。结果显示，未分化的前体脂肪细胞经DHA作用48 h，各处理组前体脂肪细胞G1期百分比均呈上升趋势，但与对照无显著性差异；分化的脂肪细胞经DHA作用24 h，40 μmol/L组细胞内COX-2 mRNA的表达量显著下降，而160 μmol/L组则显著增加。以上结果表明，DHA对大鼠前体脂肪周期无明显的调节作用，而对脂肪细胞内COX-2 mRNA的调节具有明显的浓度依赖效应。     

Pharmacokinetic study of nobiletin and tangeretin in rat serum by high-performance liquid chromatography-electrospray ionization-mass spectrometry

Nobiletin (NOB) and tangeretin (TAN), two of the main polymethoxylated flavones (PMFs) in citrus, influence a number of key biological pathways in mammalian cells. Although the impacts of NOB and TAN on glucose homeostasis and cholesterol regulation have been investigated in human clinical trials, much information is still lacking about the metabolism and oral bioavailability of these compounds in animals. In this study, NOB and TAN were administered to rats by gavage and intraperitoneal (ip) injection, and the blood serum concentrations of these compounds and their main metabolites were monitored by high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS). In addition to the administered compounds, two metabolites of TAN and eight metabolites of NOB were detected and measured over 24 h. With identical oral doses, nearly 10-fold higher absorption of NOB occurred compared to TAN. For both compounds, maximum levels of glucuronidated metabolites occurred in the blood serum at later time points (～5-8 h) compared to the earlier T(max) values for NOB and TAN. In most cases the glucuronides occurred at substantially higher concentrations than the aglycone metabolites. Low levels of NOB and TAN and their metabolites were detectable in rat blood serum even at 24 h after treatment.     

Phenolics in maize genotypes differing in susceptibility to Gibberella stalk rot (Fusarium graminearum Schwabe)

The relationship between phenolic compounds and maize pith resistance to Fusarium graminearum, the causal agent of Gibberella stalk rot, was investigated. The phenolic acid profiles in the stalks of six maize inbred lines of varying susceptibility were evaluated from silking to grain maturity. Four different fractions of phenolic compounds were extracted from inoculated and non-inoculated (control) pith tissues: insoluble cell-wall-bound, free, soluble ester-bound, and soluble glycoside-bound phenolics. Analysis by HPLC revealed that p-coumaric acid and ferulic acid were the most abundant compounds in the soluble and cell-wall-bound fractions. The quantity of free, glycoside-bound, and ester-bound phenolics in the pith was lower than the level required for the inhibition of Fusarium growth or mycotoxins production; however, significant negative correlations between diferulic acid contents in the cell walls and disease severity ratings 4 days after inoculation were found. The results indicated that future studies should focus on the levels of diferulic acids during the early infection process. Diferulates may play a role in genotypic resistance of maize to Gibberella stalk rot as preformed barriers to infection.     

Antidiabetic effects of rice hull smoke extract in alloxan-induced diabetic mice

This study investigated the protective effect of a liquid rice hull smoke extract (RHSE) against diabetes in alloxan-induced diabetic mice. Antidiabetic effects of RHSE were evaluated in both the rat insulinoma-1 cell line (INS-1) and diabetic ICR mice induced by intraperitoneal (ip) injection of alloxan. Alloxan treatment (10 mM) increased cellular reactive oxygen species (ROS) levels in the INS-1 cells, which were inversely related to cell viabilities. RHSE inhibited alloxan-induced nitric oxide (NO) generation through inhibition of inducible nitric oxide synthase (iNOS) gene expression and suppressed the inflammatory reaction in INS-1 cells through inhibition of expression of pro-inflammatory genes, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). Dietary administration of 0.5 or 1% RHSE to alloxan-induced diabetic mice caused a decrease in blood glucose and increases in both serum insulin and hepatic glycogen levels. RHSE induced decreases in glucose-6-phosphatase (G6 Pase) and phosphoenolpyruvate carboxykinase (PEPCK) levels and an increase in the glucokinase (GCK) level. These changes resulted in restoring glucose-regulating enzyme levels to control values. Histopathology showed that alloxan also induced damage of Langerhans islet cells of the pancreas and liver necrosis associated with diabetes. Oral administration of RHSE restored the islet and liver cells to normal levels. RHSE-supplemented functional food could protect insulin-producing islet cells against damage triggered by oxidative stress and local inflammation associated with diabetes.