Glutinoin, a novel antioxidative ellagitannin from Alnus glutinosa cones with glutinoic acid dilactone moiety.

The crude aqueous ethanol extract of the cones of Alnus glutinosa (L.) Gaertn. (Betulaceae; black alder, European alder) was obtained and further partitioned between water and various organic phases. The active water and butanol phases were subjected to assay-guided (DPPH) fractionation using repetitive RP HPLC until individual compounds were isolated. Their antioxidative activities, measured as SC50 values, were evaluated. The chemical structures of the isolated compounds were elucidated with the help of mass spectroscopy, (1)H NMR technique, UV spectroscopy, and chemical approaches. One novel ellagitannin, glutinoin (2), along with two known compounds, pedunculagin (1) and praecoxin D (3), were isolated and found to contribute to antioxidative activity of the A. glutinosa cones extract. The activities (SC50) of 1-3 were evaluated as 0.95 (1), 1.00 (2) and 1.01 µg mL⁻¹ (3). The scavenging effects of glutinoin (2) and praecoxin D (3) were reported for the first time.

The 1,4-naphthoquinone derivative from Pyrola rotundifolia activates AMPK phosphorylation in C2C12 myotubes.

An aqueous ethanol extract of Pyrola rotundifolia L. induced AMP-activated protein kinase (AMPK) phosphorylation in C2C12 myotubes. The bioassay-guided fractionation of the extract led to the isolation a 2-methyl-7-hydroxymethyl-1,4-naphthoquinone, or a 7'-hydroxy-chimaphilin, which showed concentration-dependent AMPK phosphorylation activity at 2.5-20 µg/ml. At a concentration of 10 µg/ml (50 µM), an approximately four-fold increase in the AMPKα(Thr¹7²) phosphorylation level was observed. The stimulatory effect of naphthoquinone on AMPK activity was comparable to that of known compounds found in natural sources that activate the AMPK signaling pathway.

Catechin-rich grape seed extract supplementation attenuates diet-induced obesity in C57BL/6J mice.

BACKGROUND: Grape seed extracts (GSE) are known to present health benefits such as antioxidative and anti-obesity effects in animal models. The purpose of this research is to determine whether the specially manufactured GSE, catechin-rich GSE (CGSE), can protect against obesity induced by a high-fat diet (HFD) and to address the mechanism underlying this effect. METHODS: The componential analysis of CGSE was performed using liquid chromatography/mass spectrometry. Oxygen consumption and the respiratory quotient were determined using 500 mg/kg CGSE administered orally for 3 days in 14- to 15-week-old male C57BL/6J mice. Nine-week-old male C57BL/6J mice were supplemented with 0.5 or 1% CGSE in a HFD for 12 weeks, and their body weight and food intake were monitored. Blood and tissue samples were collected and analyzed. RESULTS: The main polyphenol components of CGSE were catechin and epicatechin. CGSE supplementation in the HFD-induced obesity model chronically suppressed the increase in body weight and the weight of fat pads. Furthermore, CGSE improved metabolic parameter abnormalities and upregulated the fatty acid oxidation-related genes in the liver. CONCLUSIONS: These findings suggest that CGSE contains monomeric catechins in high concentrations and ameliorates HFD-induced obesity in C57BL/6J mice.

Isolation of a novel catechin from Bergenia rhizomes that has pronounced lipase-inhibiting and antioxidative properties.

An aqueous ethanol extract of Bergenia crassifolia rhizomes strongly inhibited human pancreatic lipase activity and increased scavenging of DPPH free radicals in vitro. Chromatographic separation of this extract led to isolation of the hydrolysable tannins (+)-catechin 3,5-di-O-gallate (1) and (+)-catechin 3-O-gallate (2). This is the first report of the isolation of compound 1 from plant material. This compound strongly inhibited human pancreatic lipase (with an IC(50) value of 0.42 µg/ml) and exhibited a remarkable free radical-scavenging ability (with an SC(50) value of 1.04 µg/ml). The chemical structures of 1 and 2 were elucidated using MS, NMR and chemical approaches.

Antioxidative activities of Oxindole-3-acetic acid derivatives from supersweet corn powder.

The components contributing to the antioxidative activity of supersweet corn powder (SSCP), which is commonly used in corn soup and snacks in Japan, were clarified and the effects investigated. 7-(O-ß-Glucosyloxy)oxindole-3-acetic acid (GOA) was found to be the component most strongly contributing to the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity of the 80% ethanol extract of SSCP, and the presence of its aglycone, 7-hydroxy-oxindole-3-acetic acid (HOA) was confirmed. GOA and HOA respectively contributed 35.1% and 10.5% to the DPPH radical-scavenging activity of the 80% ethanol extract of SSCP. Mice orally administered with HOA at doses of both 500 and 1500 mg/kg showed a significantly lower (p<0.05) level of thiobarbituric acid reactive substances (TBARS) in the plasma than the vehicle-treated control. These results suggest that GOA and HOA were at least partly involved in the antioxidative activity of SSCP in vitro and that HOA might have possessed antioxidative activity in vivo.

Functional loss of pAMT results in biosynthesis of capsinoids, capsaicinoid analogs, in Capsicum annuum cv. CH-19 Sweet.

Capsaicinoids are responsible for the spicy flavor of pungent peppers (Capsicum). The cultivar CH-19 Sweet is a non-pungent pepper mutant derived from a pungent pepper strain, Capsicum annuum CH-19. CH-19 Sweet biosynthesizes capsaicinoid analogs, capsinoids. We determined the genetic and metabolic mechanisms of capsinoid biosynthesis in this cultivar. We analyzed the putative aminotransferase (pAMT) that is thought to catalyze the formation of vanillylamine from vanillin in the capsaicinoid biosynthetic pathway. Enzyme assays revealed that pAMT activity catalyzing vanillylamine formation was completely lost in CH-19 Sweet placenta tissue. RT-PCR analysis showed normal mRNA transcription of the pAMT gene; however, SNP analysis of the cDNA sequence showed a T nucleotide insertion at 1291 bp in the pAMT gene of CH-19 Sweet. This insertion formed a new stop codon, TGA, that prevented normal translation of the gene, and the pAMT protein did not accumulate in CH-19 Sweet as determined using Western blot analysis. We developed a dCAPS marker based on the T insertion in the pAMT gene of CH-19 Sweet, and showed that the pAMT genotype co-segregated with the capsinoid or capsaicinoid fruit phenotype in the F(2) population. The T insertion was not found in other pungent and non-pungent Capsicum lines, suggesting that it is specific to CH-19 Sweet. CH-19 Sweet's pAMT gene mutation is an example of a nonsense mutation in a single gene that alters a secondary metabolite biosynthetic pathway, resulting in the biosynthesis of analogs. The dCAPS marker will be useful in selecting lines with capsinoid-containing fruits in pepper-breeding programs.