Analysis of Volatile Compounds from Chili Peppers and Characterization of Habanero (Capsicum chinense) Volatiles.

The Habanero pepper is characterized by its strong pungency and fruity aroma. The aim of the present study was to extract the volatile compounds of Habanero peppers, using solvent extraction and solvent-assisted flavor evaporation (SAFE) methods, and to analyze them using gas chromatography-mass spectrometry (GC-MS). The analysis detected 66 volatile compounds, including 6-methyl-(E)-4-heptenyl 3-methylbutanoate 1, which was reported previously, and 6-methyl-(E)-4-heptenyl 2-methylpropanoate 2, the corresponding butanoate 3, 2-methylbutanoate 4, and 6-methyl-(E)-4-heptenol 5, which were found in both Habanero and other peppers. 6-Methyl-(E)-4-heptenyl 3-methylbutanoate 1 and related compounds were synthesized. Furthermore, principal component analysis (PCA) and hierarchical cluster analysis (HCA) of the volatile profiles generally grouped the pepper samples by species and indicated that Habanero peppers are characterized by the presence of 6-methyl-(E)-4-heptenyl esters.

Isolation and Identification of a Volatile Compound in Habanero Pepper (Capsicum chinense).

A volatile compound was isolated from the fruit of Habanero pepper (Capsicum chinense) and related varieties and identified as 6-methyl-(E)-4-heptenyl 3-methylbutyrate by 1H and 13C NMR spectroscopy and two-dimensional NMR experiments, including HMQC, HMBC, and 1H-1H COSY. The compound has a retention index of 1387 and is one of the major volatile compounds in Habanero pepper. This compound would be useful as a new flavor.

Isolation of a reduced form of cyanidin 3-O-ß-D-glucoside from immature black soybean (Glycine max (L.) Merr.) and its reducing properties.

5,7,3',4'-Tetrahydroxyflav-2-en-3-ol 3-O-ß-D-glucoside was isolated from the seed coats of immature black soybeans (Glycine max (L.) Merr.). This compound is a reduced form of cyanidin 3-O-ß-D-glucoside (cyanidin 3-G) which was obtained by reaction with hydrochloric acid. The molecule has reducing activity for a tetrazolium derivative (WST-1) in the presence of 1-methoxy-5-methylphenazinium methylsulfate (1-methoxy PMS) in a similar manner to NADH. The seed coats of immature black soybeans also contain epicatechin as a major constituent, while cyanidin 3-G and procyanidin B2 are present at lower concentrations. Immature brown soybeans did not contain 5,7,3',4'-tetrahydroxyflav-2-en-3-ol 3-O-ß-D-glucoside, but did contain both epicatechin and procyanidin B2. Immature yellow soybeans contained none of them.

A hair growth-promoting effect of Chinese black tea extract in mice.

Chinese black tea extract (CBTE) fermented with Aspergillus sp. significantly promoted hair growth after 2 weeks of topical application in shaved 6 week-old male C3H/He mice. The hair growth-promoting effect of CBTE was potentiated synergistically by capsaicin, which has no effect on hair growth by itself. CBTE displayed an affinity for estrogen receptor (ER)α, with an IC50 value of 74.8 µg/mL. This effect of CBTE might be mediated by the ERs, since a similar effect induced by orally administered soy isoflavone, a mixture of ERs ligands, has been reported to be synergistically potentiated by capsaicin.

1,3-Capryloyl-2-arachidonoyl glycerol activates alpha-secretase activity and suppresses Abeta40 secretion in A172 cells.

Alzheimer's disease (AD) is characterized by the deposition of amyloid beta-peptide (Abeta), derived from amyloid precursor protein (APP). Membrane states, such as lipid components or membrane fluidity, are important for enzymes related to APP processing in meeting their substrates efficiently. We analyzed the effects of triglycerides combined with polyunsaturated fatty acids (PUFAs) and/or caprylic acids on APP proteolysis. Our results showed that 1,3-capryloyl-2-arachidonoyl glycerol (8A8) moderately increased alpha-secretase activity (18%) in A172 cells. beta-Secretase activity was not statistically significantly changed in HEK293 cells stably expressing BACE1. However, Abeta40 secretion decreased by 22%. Thus, we conclude that 8A8 is a useful lipid compound for activating alpha-secretase activity and suppressing Abeta40 secretion.

A novel glucosylation enzyme: molecular cloning, expression, and characterization of Trichoderma viride JCM22452 alpha-amylase and enzymatic synthesis of some flavonoid monoglucosides and oligoglucosides.

It was found that commercial cellulase preparations from Trichoderma viride showed transglucosylation activity toward (+)-catechin and (-)-epigallocatechin gallate (EGCG) using dextrin as a glucosyl donor. To isolate the enzyme exhibiting transglucosylation activity toward (+)-catechin and EGCG, the present study isolated the cDNA encoding the T. viride JCM22452 alpha-amylase homologue (TRa2), which showed high amino acid sequence identity to functionally uncharacterized alpha-amylase homologues from other ascomycetes, which also produced some (+)-catechin and EGCG glucosides. TRa2 was able to glucosylate a wide range of natural flavonoids, particularly (+)-catechin and EGCG, and to hydrolyze maltooligosaccharides (k(cat)/K(m) for maltotriose, maltotetraose, maltopentaose, maltohexaose, and maltoheptaose were 1.98, 45.2, 58.3, 97.4, and 92.6 s(-1) mM(-1), respectively) except maltose but could not transfer the monoglucosyl residue to maltooligosaccharides. By (1)H NMR and (13)C NMR, the structures of several novel glucosides obtained by commercial cellulase preparations from T. viride and TRa2 were determined as (+)-catechin 5-O-alpha-D-glucopyranoside, (+)-catechin 5-O-alpha-D-maltoside, (+)-catechin 4'-O-alpha-D-maltoside, EGCG 5-O-alpha-D-glucopyranoside, and EGCG 7-O-alpha-D-maltoside. One of these glucosides, EGCG 5-O-alpha-D-glucopyranoside, showed higher heat stability and solubility and lower astringency and astringent stimulation than its aglycon, suggesting that EGCG glucosides are functionally superior to EGCG as food additives.

Enzymatic synthesis of astaxanthin n-octanoic acid esters.

We examined the enzymatic synthesis of astaxanthin n-octanoic acid esters. Carriers for the immobilized enzyme and reaction conditions such as water content, reaction temperature, and time were examined using Candida cylindracea lipase (Lipase OF). Lipase OF) immobilized by a hydrophobic anion exchange resin (10% w/w content of lipase) gave the best yield in the esterification reaction of astaxanthin. Two milligrams of astaxanthin per 750 microL tri-n-octanoin (ca. 0.3%) was optimum because of the low solubility of tri-n-octanoin. The esters were obtained in a yield of 36.4% under the optimal reaction conditions.

Identification of 6-substituted 4-arylsulfonyl-1,4-diazepane-2,5-diones as a novel scaffold for human chymase inhibitors.

A novel series of 6-substituted 4-sulfonyl-1,4-diazepane-2,5-diones were designed, synthesized and evaluated as human chymase inhibitors. Structure-activity relationship studies led to the identification of a potent inhibitor, (6S)-6-(5-chloro-2-methoxybenzyl)-4-[(4-chlorophenyl)sulfonyl]-1,4-diazepane-2,5-dione, with an IC(50) of 0.027 microM.

Selectivity of fatty acid ligands for PPARalpha which correlates both with binding to cis-element and DNA binding-independent transactivity in Caco-2 cells.

It is thought that peroxisome proliferator-activated receptor alpha (PPARalpha) is a major regulator for fatty acid metabolism. Long-chain fatty acids have been shown to induce expression of the genes related to fatty acid metabolism through PPARalpha. However, it is unclear whether the intensity of PPARalpha activation is different among various fatty acids. In this study, we compared various fatty acids in the capability of PPARalpha activation by differential protease sensitivity assay (DPSA), electrophoretic mobility shift assay and GAL4-PPAR chimera reporter assay in intestinal cell line, Caco-2. DPSA revealed that polyunsaturated fatty acids of 18 to 20 carbon groups with 3-5 double bonds strongly induced a PPARalpha conformational change. The ligand-induced changes in the sensitivity to protease corresponded to the enhancement of the binding of PPARalpha-RXRalpha heterodimer to the PPAR-response element (PPRE). The GAL4-PPAR chimera reporter assay revealed that the DNA binding-independent transactivity of PPARalpha was induced by various fatty acids with a wide spectrum of intensity which correlated with the conformational change of PPARalpha. These results suggest that PPARalpha has greater selectivity to certain types of polyunsaturated fatty acids, and that the ligand-induced conformational change of PPARalpha leads to parallel increases in both DNA binding to the PPAR-response element and the DNA binding-independent transactivity.

Evaluation of the hypolipemic property of Camellia sinensisVar. ptilophylla on postprandial hypertriglyceridemia.

A naturally decaffeinated tea, Camellia sinensis var. ptilophylla (cocoa tea), has long been popular in southern China as a healthy beverage. Our experiments indicate that a single oral administration of 500 mg/kg of cocoa tea extract suppresses increases in plasma triacylgycerol (TG) levels when fed with 5 mL/kg of olive or lard oil in mice and that the inhibition rates are 22.9% and 31.5%, respectively, compared with controls. Under the same condition, cocoa tea extract did not affect the level of plasma free fatty acid. Likewise, the extract reduced the lymphatic absorption of lipids at 250 and 500 mg/kg. Also, cocoa tea extract and polyphenols isolated from cocoa tea inhibit pancreatic lipase. These findings suggest that cocoa tea has hypolipemic activity, which may be due to the suppression of digestive lipase activity by the polyphenols contained within the tea.

A novel vitamin C analog, 2-O-(beta-D-Glucopyranosyl)ascorbic acid: examination of enzymatic synthesis and biological activity.

2-O-(beta-D-Glucopyranosyl)ascorbic acid (AA 2 beta G) isolated from a popular traditional Chinese food (Lycium fruit) was synthesized using cellulase derived from Trichoderma sp. with cellobiose as a glucose donor. 6-O-(beta-D-Glucopyranosyl)ascorbic acid as well as AA 2 beta G was also synthesized in this reaction. The vitamin C activity of AA 2 beta G was also evaluated using inherently scorbutic (osteogenic disorder Shionogi [ODS]) rats. The rats were fed vitamin C-deficient food and water containing AA 2 beta G for 21. AA 2 beta G supported their growth and the level of vitamin C in tissues was moderately maintained. The vitamin C level in some tissues depended on the hydrolytic activity of AA 2 beta G (beta-glucosidase activity) although the correlation was not statistically significant (P=0.08). The results indicate that AA 2 beta G has pro-vitamin C activity.

Creation of Rhizopus oryzae lipase having a unique oxyanion hole by combinatorial mutagenesis in the lid domain.

Combinatorial libraries of the lid domain of Rhizopus oryzae lipase (ROL; Phe88Xaa, Ala91Xaa, Ile92Xaa) were displayed on the yeast cell surface using yeast cell-surface engineering. Among the 40,000 transformants in which ROL mutants were displayed on the yeast cell surface, ten clones showed clear halos on soybean oil-containing plates. Among these, some clones exhibited high activities toward fatty acid esters of fluorescein and contained non-polar amino acid residues in the mutated positions. Computer modeling of the mutants revealed that hydrophobic interactions between the substrates and amino acid residues in the open form of the lid might be critical for ROL activity. Based on these results, Thr93 and Asp94 were further combinatorially mutated. Among 6,000 transformants, the Thr93Thr, Asp94Ser and Thr93Ser, Asp94Ser transformants exhibited a significant shift in substrate specificity toward a short-chain substrate. Computer modeling of these mutants suggested that a unique oxyanion hole, which is composed of Thr85 Ogamma and Ser94 Ogamma, was formed and thus the substrate specificity was changed. Therefore, coupling combinatorial mutagenesis with the cell surface display of ROL could lead to the production of a unique ROL mutant.

Effects of oolong tea on plasma antioxidative capacity in mice loaded with restraint stress assessed using the oxygen radical absorbance capacity (ORAC) assay.

In the present study, we investigated the antioxidative effect of oolong tea in vitro and in vivo using the oxygen radical absorbance capacity (ORAC) assay. An oolong tea extract, catechin and related compounds suppressed the oxidation of fluorescence induced by AAPH in a dose-dependent manner, that is, they prolonged the antioxidant time in vitro. Oral administration of the oolong tea extract to mice treated with restraint stress increased ORAC activity in plasma as compared with a stress control group. The extract also increased plasma vitamin C levels, and there was a good relationship between ORAC activity and the vitamin C level in plasma. The elevation of plasma ORAC and vitamin C level may have been related to the stress-relieving effect of oolong tea. These effects are probably due to the antioxidative properties of the tea. Thus, these findings suggested that oolong tea has beneficial effects on health related to its antioxidative action.

2-O-(beta-D-Glucopyranosyl)ascorbic acid, a novel ascorbic acid analogue isolated from Lycium fruit.

A novel stable precursor of ascorbic acid (vitamin C), 2-O-(beta-D-glucopyranosyl)ascorbic acid, was isolated from both the ripe fresh fruit and dried fruit of Lycium barbarum L., a plant of the Solanaceae family. The chemical structure was inferred by instrumental analyses and confirmed by chemical synthesis. The dried fruit of Lycium barbarum L. contained ca. 0.5% of it, which is comparable to the ascorbic acid content of fresh lemons. It increased the blood ascorbic acid by oral administration to rats, and it was also detected in blood from the portal vein.

Involvement of mast cell chymase in bleomycin-induced pulmonary fibrosis in mice.

The possible role of mast cell chymase in organ fibrosis was examined using a bleomycin-induced pulmonary fibrosis model in mice. Intratracheal injection of bleomycin to mice significantly increased not only hydroxyproline content but also chymase activity in the lung. Administration of a chymase inhibitor SUN C8077 (7-chloro-3-(3-amynophenyl) quinazoline-2, 4-dione methanesulfonate) dose-dependently reversed the bleomycin-induced increase in hydroxyproline content as well as chymase activity in the lung. Human chymase digested latent transforming growth factor-beta1 (TGF-beta1) to form mature TGF-beta1 in vitro, which was inhibited by SUN C8077. Human chymase, on the other hand, failed to stimulate DNA synthesis of human lung fibroblasts CCD-8Lu and LL97A. Taken together, it is suggested that mast cell chymase might participate in the pathogenesis of pulmonary fibrosis, and that the chymase-induced fibrosis might be mediated at least in part by TGF-beta1. Chymase inhibitor may be promising for treatment of pulmonary fibrosis in humans.

Influence of histamine in a liver injury model induced by Propionibacterium acnes and lipopolysaccharide.

In normal mice, plasma histamine levels were 29.4+/-10.1 pmol/ml. When 0.1 microg of lipopolysaccharide (LPS) was intravenously injected into Propionibacterium acnes (P. acnes)-primed ICR mice, histamine levels increased remarkably to 61.2+/-15.9 pmol/ml (p<0.001). An increase was also observed in liver tissues. Oral administration of histidine at 200 mg/kg once daily for 5 d before intravenous LPS injection increased the plasma alanine aminotransferase (ALT) activity to 2936.5+/-356.3 IU/l, a significant change compared with the controls (2244.8+/-425.5 IU/l, p<0.05). The 24 h survival rate after LPS injection was 72.7% in the mice treated with 50 mg/kg of ranitidine, in contrast with 50% in the control group although the treatment did not significantly decrease the plasma ALT activity. On the other hand, 50 mg/kg of pyrilamine significantly reduced plasma ALT activity (p<0.001). The results suggested that histamine levels are related to hepatic damage in the P. acnes plus LPS induction of liver injury.

Hypoglycemic action of Cyclocarya paliurus (Batal.) Iljinskaja in normal and diabetic mice.

This study examined the hypoglycaemic activity of Cyclocarya paliurus (Batal.) Iljinskaja (C. paliurus) in ICR mice by oral glucose tolerance testing. The blood glucose level was significantly lower in the C. paliurus extract treatment group than in the control group after animals were given sucrose. This difference was not observed following the administration of glucose. We demonstrated that the chronological change in the level of blood glucose in genetically hyperglycemic obese KK-Ay mice is significantly lower when C. paliurus extract is administered daily for three weeks. An in vitro study showed that C. paliurus inhibits alpha-glucosidase, a disaccharide-degrading enzyme in the small intestinal mucosa, leading to a decrease in the absorption of glucose into the blood and a subsequent lowering of the blood glucose level.

Inhibitory effect of oolong tea on the oxidative state of low density lipoprotein (LDL).

In the present study, we investigated the anti-oxidant activity of oolong tea in an oxidation model using human low-density lipoprotein (LDL). Oolong tea suppressed the oxidation of LDL induced by 2-2'-azobis 4-methoxy-2,4-dimethyvaleronitrile (V70) in a dose-dependent manner, that is, it prolonged the lag time to 114.3%, 138% and 199.9% as compared with the control group at 0.5 microg/ml, 1.0 microg/ml, and 2.5 microg/ml, respectively. We also determined the scavenging effect of oolong tea on active oxygen radicals using the electron spin resonance (ESR) technique with 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin trapping agent. The intensity of the ESR signals for the DMPO-OOH adduct formed by the hypoxanthine/xanthine oxidase reaction system with DMPO decreased in the presence of oolong tea. The IC(50) of oolong tea was 19.9 microg/ml. These findings suggested that oolong tea has beneficial effects on health related to its anti-oxidative action.

Hypolipemic effect of Cyclocarya paliurus (Batal) Iljinskaja in lipid-loaded mice.

We investigated the inhibitory effect of Cyclocarya paliurus (Batal.) Iljinskaja (C. paliurus) extract on postprandial hyperlipemia in mice. A single oral administration of C. paliurus extract (250 mg/kg) suppressed an increase in plasma triacylgycerol (TG) levels when fed with 5 ml/kg of lard and olive oil. The inhibition rates were 28.6% and 24.1%, respectively, but free fatty acid (FFA) levels in plasma were not significantly affected as compared with control group mice. In addition, C. paliurus extract showed inhibitory activity toward pancreatic lipase, a key enzyme of dietary TG absorption, with an IC(50) of 9.1 microg/ml in vitro. Our results suggested that the hypolipemic action of C. paliurus extract was probably interrelated with suppression of the activity of digestive lipase, and as a result, the blood lipid level was reduced.

Effects of oolong tea on metabolism of plasma fat in mice under restraint stress.

We investigated the effects of oolong tea on the basic metabolism of plasma lipids in mice under restraint stress. When a lipid emulsion (Intralipid 20%; a lipid emulsion containing 20% soybean oil) was injected intravenously into mice, the restraint stress prolonged the half-life (T 1/2) of elimination for plasma triglyceride (TG) from 28.7 to 55.5 min. The elimination rate per minute was 48.2% in stressed mice with the rate in starved control mice as 100%. Therefore, TG metabolism was disrupted by the stress, and the use of TG as an energy source decreased. We found that the metabolism of lipids significantly response to the restrained stress in the present study. Plasma TG was 515.9 +/- 29.9mg/dl 35min after Intralipid administration in control stressed mice, 478.7 +/- 26.7 mg/dl in the stressed group given caffeine 100 mg/kg of body weight, and 418.3 +/- 18.4 mg/dl in the stressed group given 1,000 mg/kg oolong tea, an improvement by 7.2% and 18.9%, respectively, with the value for the untreated control group. The intake of oolong tea alleviated the stress-induced decrease in the rate of blood lipid metabolism; this effect may have arisen from some non-specific stress-relieving property of the tea or from acceleration of lipid metabolism by properties of polyphenols, etc. in tea. Oolong tea had anti-stress effects on plasma TG metabolism, and the effects did not depend on caffeine.