PHYTOCHEMICAL AND ANTIOXIDANT CHARACTERIZATION OF THINNED IMMATURE CITRUS UNSHIU FRUITS

Objective: We aimed to evaluate the characterization of thinned immature Citrus unshiu fruits with regard to their phytochemical profile and antioxidant capacity.Methods: Determination of total phenolic, flavonoid, and carotenoid and ascorbic acid contents was done by UV-Visible spectrophotometry, whereas UPLC-mass detection was used for the analysis of individual flavanone (naringin, hesperidin, hesperetin, neohesperedine and narirutin) and flavonol (rutin). In addition, free radicals (DPPH, O2-, H2O2 and NO) scavenging assays were used to determine the antioxidant capacity.Results: Naringin, hesperidine, neohesperedine and narirutin were the main flavanones in all thinned immature Citrus unshiu fruits. The contents of total phenolic, flavonoid and carotenoid were more prevalent in immature fruits than the level found in mature fruits. All thinned immature Citrus unshiu fruits possess an evident antioxidant capacity. The immature Citrus extract concentrations providing 50% inhibition (IC50) for free radicals; 1.2-1.49 mg/ml for DPPH, 1.03-1.46 mg/ml for superoxide, 1.95-3.43 mg/ml for hydrogen peroxide and 1.64-3.45 mg/ml for nitric oxide was lower than those of mature Citrus extracts.Conclusion: Thinned immature Citrus unshiu fruits could be an economic and readily accessible source of natural antioxidants and as a possible food and pharmaceutical supplement

Effect of dose and dosing rate on the mutagenesis of nitric oxide in supF shuttle vector

Purpose: To determine how the dose and rate of NO• treatment affects mutagenic responses.Methods: Shuttle vector pSP189 was used to determine the genotoxicity resulting from in vitro exposure to NO• using three delivery methods (reactor and Transwell co-culture systems, and NO• donor sodium nitroprusside), followed by plasmid replication in bacteria MBL50 and human AD293 cells.Results: When exposed to preformed 100% NO• for 3 h or 1% NO• for 35 h using a reactor system, a cumulative dose of 1260 μM × min reduced AD293 cell viability by 46 and 18% and increased mutation frequencies (MFs) 1.9- and 5.3-fold higher than argon control, respectively. Roughly 5-fold increase in MF of the supF gene of AD293 cells co-cultivated with macrophages stimulated with IFN-γ/LPS was also observed. When AD293 cells were treated by SNP, DNA strand breaks were induced and MFs were increased in a dose-dependent manner.Conclusion: These results provide important clues to how dose and dosing rate of introducing NO• may contribute to potential genotoxicity resulting from NO• formation in vivo.Keywords: AD293 cells, Delivery method, Genotoxicity, Nitric oxide, supF Gene of pSP189 shuttle vecto

Apoptotic properties of Citrus sudachi Hort, ex Shirai (Rutaceae) extract on human A549 and HepG2 cancer cells

Purpose: To investigate whether Citrus sudachi harvested at two stages of maturity can induce toxicity in a cell-specific manner and to determine the possible  mechanisms of Citrus sudachi-induced cytotoxic responses in two types of cancer cells (human lung adenocarcinoma A549 and hepatocellular carcinoma HepG2 cells) and two normal cell lines (lung 16HBE140- and liver CHANG cells).Methods: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and annexin V/propidium iodidle assay were used to test the antiproliferative activity and apoptosis of methanol extract of Citrus sudachi, respectively. Griess reaction and reverse transcriptase-polymerase chain reaction (RT-PCR) were carried out to evaluate nitric oxide (NO•) production and the mRNA levels of inhibitors of apoptosis (IAP).Results: Citrus sudachi exerted cytotoxicity in a time-dependent manner in cancer cells which increased with increase in maturity but did not affect normal cells. Citrus sudachi was found to induce accumulation of cells in the sub-G1 cell cycle phase, fragmentation of DNA and cell death with characteristics of apoptosis, in both types of cancer cells. Moreover, Citrus sudachi upregulated cellular NO• produced by activation of nitric oxide synthase (NOS), while it suppressed the levels of IAP mRNA in both types of cancer cells.Conclusion: The results obtained suggest that Citrus sudachi induces apoptosis in A549 and HepG2 cells, which may be mediated by NO•. There is need for further studies on the role of Citrus sudachi in cancer treatment.Keywords: Apoptosis, Citrus sudachi, Human lung and liver cancer cells, Inhibitors of apoptosis, Nitric oxid

Effect of extraction solvents on antioxidant and skin-whitening potentials of defatted Camellia seed cakes

Defatted Camellia japonica L. seed cake is an important byproduct during the manufacture of Camellia seed oil. The present study evaluated the influence of two extraction solvents on the total contents of phenol and flavonoid, antioxidant activity and skin-whitening effect capable of inhibiting the biosynthesis of melanin of defatted Camellia seed cakes, a byproduct from Camellia oil production. The antioxidant capacities of 100% methanol and 70% ethanol extracts were analysed using radical scavenging (1,1-diphenyl-2-picrylhydrazyl, O2-, H2O2 and NO), SOD-like, ferrous ion chelating and reducing power assays. The total phenolic and flavonoid contents were further determined by the Folin-Ciocalteu method. Moreover, intracellular antityrosinase activity and melanin contents were evaluated in human malignant melanoma cells (SK mel-100). Ethanol extracts of defatted Camellia seed cake extracts exhibited higher phenolic (4097 mg gallic acid equivalents/100 g) and flavonoid (2899 mg rutin equivalents/100 g) contents with higher superoxide (IC50 = 1.9 mg/mL), nitric oxide (IC50 =1.6 mg/mL) radical scavenging, ferrous ion chelating (IC50 = 2.9 mg/mL) and reducing power (IC50 = 1.8 mg/mL) activities than those of methanol. These ethanol extracts also evidenced more effective inhibitory activities of tyrosinase and melanin synthesis than methanol extracts. Therefore, the present results demonstrated that defatted Camellia seed cakes could be a valuable source of antioxidative and whitening ingredients, and ethanol was more efficient in extracting antioxidants and bioactive compounds than methanol

Onion peel extracts ameliorate hyperglycemia and insulin resistance in high fat diet/streptozotocin-induced diabetic rats

<p>Abstract</p> <p>Background</p> <p>Quercetin derivatives in onions have been regarded as the most important flavonoids to improve diabetic status in cells and animal models. The present study was aimed to examine the hypoglycemic and insulin-sensitizing capacity of onion peel extract (OPE) containing high quercetin in high fat diet/streptozotocin-induced diabetic rats and to elucidate the mechanism of its insulin-sensitizing effect.</p> <p>Methods</p> <p>Male Sprague-Dawley rats were fed the AIN-93G diet modified to contain 41.2% fat and intraperitoneally injected with a single dose of streptozotocin (40 mg/kg body weight). One week after injection, the rats with fasting blood glucose levels above 126 mg/dL were randomly divided into 4 groups to treat with high fat diet containing 0 (diabetic control), 0.5, or 1% of OPE or 0.1% quercetin (quercetin equivalent to 1% of OPE) for 8 weeks. To investigate the mechanism for the effects of OPE, we examined biochemical parameters (insulin sensitivity and oxidative stresses) and protein and gene expressions (pro-inflammatory cytokines and receptors).</p> <p>Results</p> <p>Compared to the diabetic control, hypoglycemic and insulin-sensitizing capability of 1% OPE were demonstrated by significant improvement of glucose tolerance as expressed in incremental area under the curve (<it>P </it>= 0.0148). The insulin-sensitizing effect of OPE was further supported by increased glycogen levels in liver and skeletal muscle (<it>P </it>< 0.0001 and <it>P </it>= 0.0089, respectively). Quantitative RT-PCR analysis showed increased expression of insulin receptor (<it>P </it>= 0.0408) and GLUT4 (<it>P </it>= 0.0346) in muscle tissues. The oxidative stress, as assessed by superoxide dismutase activity and malondialdehyde formation, plasma free fatty acids, and hepatic protein expressions of IL-6 were significantly reduced by 1% OPE administration (<it>P </it>= 0.0393, 0.0237, 0.0148 and 0.0025, respectively).</p> <p>Conclusion</p> <p>OPE might improve glucose response and insulin resistance associated with type 2 diabetes by alleviating metabolic dysregulation of free fatty acids, suppressing oxidative stress, up-regulating glucose uptake at peripheral tissues, and/or down-regulating inflammatory gene expression in liver. Moreover, in most cases, OPE showed greater potency than pure quercetin equivalent. These findings provide a basis for the use of onion peel to improve insulin insensitivity in type 2 diabetes.</p

2,9,16,19,22,25-Hexaoxatetra­cyclo­[24.4.0.24,7.010,15]dotriaconta-1(26),4,6,10(15),11,13,27,29,31-nona­ene

The title 22-crown-6 unit, C26H28O6, comprising of three benzo groups and triethyl­ene glycol, was prepared by the reaction of α,α′-dibromo-p-xylene with 1,8-bis­(2-hydroxy­phen­oxy)-3,6-dioxaoctane in the presence of Cs2CO3 with tetra­hydro­furan (THF) and recrystallized from dichloro­methane–hexane (1:20 v/v) at room temperature. In the mol­ecular structure, two O atoms of the central ethyl­ene glycol in the triethyl­ene glycol unit exhibit exo conformations due to intra­molecular C—H⋯O inter­actions. A number of C—H⋯O and C—H⋯π inter­molecular inter­actions contribute to the stabilization of the crystal packing

Binding of the influenza A virus NS1 protein to PKR mediates the inhibition of its activation by either PACT or double-stranded RNA

AbstractA major component of the cellular antiviral system is the latent protein kinase PKR, which is activated by binding to either double-stranded RNA (dsRNA) or the cellular PACT protein. Activated PKR phosphorylates the translation initiation factor eIF2, thereby inhibiting viral and cellular protein synthesis and virus replication. To evade the antiviral effects of PKR, many viruses, including influenza A virus, have evolved multiple mechanisms. For influenza A virus, the non-structural (NS1A) protein plays a major role in blocking activation of PKR during virus infection. The mechanism by which the NS1A protein inhibits PKR activation in infected cells has not been established. In the present study, we first carried out a series of in vitro experiments to determine whether the NS1A protein could utilize a common mechanism to inhibit PKR activation by both PACT and dsRNA, despite their different modes of activation. We demonstrated that the direct binding of the NS1A protein to the N-terminal 230 amino acid region of PKR can serve as such a common mechanism and that this binding does not require the RNA-binding activity of the NS1A protein. The lack of requirement for NS1A RNA-binding activity for the inhibition of PKR activation in vivo was established by two approaches. First, we showed that an NS1A protein lacking RNA-binding activity, like the wild-type (wt) protein, blocked PKR activation by PACT in vivo, as well as the downstream effects of PKR activation in cells, namely, eIF2 phosphorylation and apoptosis. In addition, we demonstrated that PKR activation is inhibited in cells infected with a recombinant influenza A virus expressing NS1A mutant protein that cannot bind RNA, as is the case in cells infected with wild-type influenza A virus