Comparison of Nutritional Composition and Antioxidant Properties of Pulverized and Unutilized Portions of Waxy Barley.

To promote the use of waxy barley bran, an underutilized resource, samples of waxy barley were divided into three parts: polished waxy barley powder (PWBP), inner bran layer powder (IBLP), and outer bran layer powder (OBLP). The color and appearance, general properties, minerals, vitamins, ß-glucan, antioxidant properties, and aroma of each part were compared. In terms of appearance and color, IBLP and OBLP appeared more yellow than PWBP; general components that were more abundant in IBLP and OBLP compared with PWBP were protein, fat, and ash. IBLP and OBLP had characteristically high values of Mg and Zn, monounsaturated and polyunsaturated fatty acids, vitamin B1, total polyphenol content, H-ORAC, and DPPH. In particular, the vitamin B1 content of OBLP was approximately 10 times higher than that of PWBP, and Mg and Zn content was more than five times higher than in PWBP. The ß-glucan content of IBLP and OBLP was lower than that of PWBP, but relatively high. GC-MS analysis revealed that hexanal was the aroma component common to all three samples, and the peak areas were in the order of PWBP > OBLP > IBLP.

Low temperature modulates natural peel degreening in lemon fruit independently of endogenous ethylene.

Peel degreening is an important aspect of fruit ripening in many citrus fruit, and previous studies have shown that it can be advanced by ethylene treatment or by low-temperature storage. However, the important regulators and pathways involved in natural peel degreening remain largely unknown. To determine how natural peel degreening is regulated in lemon fruit (Citrus limon), we studied transcriptome and physiochemical changes in the flavedo in response to ethylene treatment and low temperatures. Treatment with ethylene induced rapid peel degreening, which was strongly inhibited by the ethylene antagonist, 1-methylcyclopropene (1-MCP). Compared with 25 ºC, moderately low storage temperatures of 5-20 °C also triggered peel degreening. Surprisingly, repeated 1-MCP treatments failed to inhibit the peel degreening induced by low temperature. Transcriptome analysis revealed that low temperature and ethylene independently regulated genes associated with chlorophyll degradation, carotenoid metabolism, photosystem proteins, phytohormone biosynthesis and signalling, and transcription factors. Peel degreening of fruit on trees occurred in association with drops in ambient temperature, and it coincided with the differential expression of low temperature-regulated genes. In contrast, genes that were uniquely regulated by ethylene showed no significant expression changes during on-tree peel degreening. Based on these findings, we hypothesize that low temperature plays a prominent role in regulating natural peel degreening independently of ethylene in citrus fruit.

Nobiletin and related polymethoxylated flavones bind to and inhibit the nuclear export factor Exportin-1 in NK leukemia cell line KHYG-1.

Polymethoxylated flavones (PMFs) are naturally occurring compounds that have biological effects on many cell types. We previously demonstrated that PMFs such as nobiletin potentiate the cytolytic activity of the human leukemic natural killer cell line KHYG-1 and increased level of the cytotoxic protein granzyme B (GrB) and the cytokine interferon-γ (IFN-γ). However, the precise mechanisms by which this occurs remain to be elucidated. In this study, we sought to identify and investigate the function of intracellular primary targets of the PMFs in KHYG-1 cells. Using affinity purification and mass spectrometry, we identified that 3'-hydroxy-4',5,6,7-tetramethoxyflavone (TMF) binds to the nuclear export factors Exportin-1 and -2 (XPO1 and XPO2) as TMF-binding proteins and demonstrated that nobiletin competes with TMF for XPO1 binding, suggesting that nobiletin also binds to XPO1. Treatment of KHYG-1 cells with leptomycin B, a specific XPO1 inhibitor, increased the expression of GrB and IFN-γ but did not potentiate lysis of specific target cells, suggesting that the cargo of XPO1 contributes to the expression of cytolytic genes but that this alone is insufficient to enhance cytolysis. Consistent with this, nobiletin and related PMFs induced the nuclear retention of NF-κB, a transcription factor that promotes GrB and IFN-γ expression. PMFs also induced the nuclear retention of the tumor suppressor protein p53, a known XPO1 cargo protein, resulting in KHYG-1 cell cycle arrest. Collectively, these results suggest that PMFs modulate KHYG-1 function, at least in part, by inhibiting XPO1.

Effects of high ß-glucan barley on visceral fat obesity in Japanese individuals: A randomized, double-blind study.

OBJECTIVE: The aim of this study was to investigate whether a diet in which high ß-glucan barley was substituted for rice would reduce visceral fat obesity in Japanese individuals. METHODS: A randomized, double-blind, placebo-controlled intervention study was conducted with 100 Japanese individuals with waist circumference (WC) ≥85 cm for men or ≥90 cm for women and body mass index (BMI) ≥24 kg/m2. Participants were randomly assigned to consume a mixture of rice and either high ß-glucan barley (test group, 4.4 g/d) or ß-glucan-free barley (placebo group) for 12 wk. Blood samples and computed tomography scans were obtained before and after the trial. RESULTS: Both groups showed decreases in body weight and BMI, and these changes were significantly greater in the test group. WC and visceral fat area (VFA) significantly decreased in both groups (VFA: -10.7 cm2 in the test group; -6.8 cm2 in the placebo group). These changes did not differ significantly between the groups. However, a subgroup analysis of participants with VFA ≥100 cm2 showed a significant decrease in the test group, and this decrease was significantly greater than in the placebo group. CONCLUSIONS: The intake of high ß-glucan barley led to significant and safe reductions in VFA, body weight, BMI, and WC in individuals with visceral fat obesity with VFA ≥100 cm2. Barley high in ß-glucan may contribute to preventing visceral fat obesity.

Polymethoxylated flavones potentiate the cytolytic activity of NK leukemia cell line KHYG-1 via enhanced expression of granzyme B.

Polymethoxylated flavones (PMFs) are found in the peel tissues of some citrus species. Here, we report that PMFs, such as nobiletin, potentiate the cytolytic activity of KHYG-1 natural killer (NK) leukemia cells. Nobiletin markedly enhanced the expression of granzyme B, a serine protease that plays critical roles in the cytolytic activity of NK cells. The potentiated cytolytic activity induced by nobiletin was canceled by the granzyme B inhibitor Z-AAD-CMK. Nobiletin also increased the levels of phosphorylated CREB, ERK1/2, and p38 MAPK in KHYG-1 cells, which are known to participate in NK cell function. Inhibition of an upstream kinase of ERK1/2 failed to reduce the granzyme B expression and KHYG-1 cytolytic activity. Meanwhile, inhibition of p38 MAPK attenuated both granzyme B expression and KHYG-1 cytolytic activity. These results suggest that the primary role of nobiletin in KHYG-1 cytolytic activity lies in upregulation of granzyme B expression, at least in part, mediated through p38 MAPK function.

Low-temperature-modulated fruit ripening is independent of ethylene in 'Sanuki Gold' kiwifruit.

Fruit ripening in response to treatments with propylene, 1-methycyclopropene (1-MCP), and low temperature was characterized in 'Sanuki Gold' kiwifruit, Actinidia chinensis Planch. Propylene treatment immediately induced rapid fruit softening, increased AC-PG (polygalacturonase) and AC-EXP (expansin) mRNA accumulation, and stimulated an increase in the soluble solid concentration (SSC) and a decrease in titratable acidity (TA). After 3 d exposure to propylene, ethylene production and AC-PL (pectate lyase) mRNA accumulation were observed. 1-MCP treatment after 24 h exposure to propylene eliminated AC-PG mRNA accumulation and suppressed continued changes in SSC and TA. Application of 1-MCP at the start of the treatment, followed by continuous propylene exposure, markedly delayed fruit softening, and the expression of the cell wall-modifying genes, and changes in the SSC and TA, indicating that kiwifruit become insensitive to ethylene at least for 3 d following 1-MCP exposure. Surprisingly, significant fruit softening, mRNA accumulation of AC-PG, AC-PL, and AC-EXP, and decreased TA were observed without ethylene production in intact fruit stored at low temperature for 1 month, but not in fruit stored at room temperature. Repeated 1-MCP treatments (twice a week) failed to inhibit the changes that occurred in low temperature storage. These observations indicate that low temperature modulates the ripening of kiwifruit in an ethylene-independent manner, suggesting that kiwifruit ripening is inducible by either ethylene or low temperature signals.

A fraction of unripe kiwi fruit extract regulates adipocyte differentiation and function in 3T3-L1 cells.

Adipocyte dysfunction is strongly associated with the development of insulin resistance and diabetes, and regulation of adipogenesis is important in prevention of diabetes. We prepared a 100% methanol fraction of methanolic extract from unripe kiwi fruit (Actinidia deliciosa), designated KMF (kiwi fruit methanol fraction). When applied to 3T3-L1 preadipocyte cells, KMF promoted adipocyte differentiation, increased glycerol-3-phosphate dehydrogenase (GPDH) activity, and increased triglyceride (TG) content. KMF markedly increased mRNA expression of peroxisome proliferator-activated receptor gamma (PPARgamma)-the master adipogenic transcription factor-and its target genes. Moreover, KMF increased mRNA expression and protein secretion of adiponectin, whereas mRNA expression and secretion of monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) were decreased. Compared with troglitazone, KMF decreased the production of reactive oxygen species (ROS) and nuclear factor-kappaB (NFkappaB) activation. Glucose uptake was stimulated by KMF in differentiated 3T3-L1 adipocytes. Taken together, these results indicate that KMF may exert beneficial effects against diabetes via its ability to regulate adipocyte differentiation and function.

Flavanone exhibits PPARgamma ligand activity and enhances differentiation of 3T3-L1 adipocytes.

Flavanones are class of polyphenolic compounds, some of which are found in foods and provide health benefits. In this study, we show that flavanone significantly enhances differentiation of 3T3-L1 preadipocytes. During adipogenesis, flavanone enhanced expression of genes and accumulation of proteins that are involved in adipocyte function. Some reports have indicated that flavanone inhibits proliferation of mammalian cells, and down-regulates expression of growth-related proteins. Such proteins include phosphorylated ERK1/2, cyclins, and Cdks that are important for an early event in adipogenesis, mitotic clonal expansion (MCE). We demonstrated that flavanone did not inhibit MCE or expression of MCE-related proteins, except for a modest inhibition of cyclin D1 expression. Using luciferase reporter assays, we found that flavanone acted as a peroxisome proliferator-activated receptor gamma (PPARgamma) ligand in a dose-dependent manner. Together, our results suggest that flavanone enhances adipogenesis, at least in part, through its PPARgamma ligand activity.

Sakuranetin induces adipogenesis of 3T3-L1 cells through enhanced expression of PPARgamma2.

Sakuranetin (5,4'-dihydroxy-7-methoxyflavone) belongs to the flavanone class of polyphenols predominantly known as phytoalexin in rice plant. In this study, we demonstrate that sakuranetin strongly induces differentiation of 3T3-L1 preadipocytes, as evidenced by increased triglyceride accumulation and glycerol-3-phosphate dehydrogenase (GPDH) activity. In addition, even in the absence of adipogenic hormonal stimuli, sakuranetin strongly induced adipogenesis and expression of genes that are critical for the adipocytes phenotype. Time-course analyses indicated that sakuranetin induces PPARgamma2 expression without prior induction of C/EBPbeta, a transcriptional regulator of PPARgamma2 in adipogenesis. In 3T3-L1 preadipocytes, the transcriptional factors GATA-2 and GATA-3 are known to down-regulate adipogenesis by direct binding to the C/EBPbeta protein and to the GATA-binding site on the PPARgamma2 promoter. We found that sakuranetin significantly reduced the expression of GATA-2. Moreover, we observed that sakuranetin stimulated glucose uptake in differentiated 3T3-L1 adipocytes. These results suggest that sakuranetin may contribute to maintain glucose homeostasis in animals.

Nobiletin enhances differentiation and lipolysis of 3T3-L1 adipocytes.

Nobiletin is a polymethoxylated flavone found in certain citrus fruits. Here we demonstrate that nobiletin enhance differentiation of 3T3-L1 preadipocytes. Nobiletin dose-dependently increased accumulation of lipid droplets in adipocytes. Quantitative RT-PCR analyses indicated that nobiletin increased the expression of genes critical for acquisition of the adipocyte phenotype. Some of them were known peroxisome proliferator activated receptor gamma (PPARgamma) targets and PPARgamma itself, however, nobiletin did not exhibit PPARgamma ligand activity. We observed the expression of CCAAT/enhancer binding protein beta (C/EBPbeta), a transcription factor for PPARgamma, was increased by nobiletin. The activation of cAMP-responsive element binding protein (CREB) and extracellular signal-regulated kinase (ERK), which play important roles in C/EBPbeta expression were also potentiated by nobiletin. Furthermore, nobiletin stimulated lipolysis in differentiated adipocytes, which is known to be stimulated by cAMP pathway. These results suggested that nobiletin enhanced both differentiation and lipolysis of adipocyte through activation of signaling cascades mediated by cAMP/CREB.

Sennidin stimulates glucose incorporation in rat adipocytes.

A novel small molecule compound which exerts insulin mimetic is desirable. Dozens of natural products that have quinone, naphthoquinone, or anthraquinone structure, were tested by a glucose incorporation assay. We found that sennidin A, anthraquinone derivative, stimulated glucose incorporation to near level of maximal insulin-stimulated and sennidin B, a stereoisomer of sennidin A, also stimulated, but the activity of sennidin B was lower than sennidin A. Sennidin A-stimulated glucose incorporation was completely inhibited by wortmannin. Sennidin A did not induce tyrosine phosphorylation of insulin receptor (IR) and insulin receptor substrate-1 (IRS-1), but induced phosphorylation of Akt and glucose transporter 4 (GLUT4) translocation. Our results suggest that in rat adipocytes, sennidin A stimulates glucose incorporation in the phosphatidylinositol 3-kinase (PI3K)- and Akt-dependent, but in the IR/IRS1-independent manner.