Synthesis of conjugated linoleic acid by human-derived Bifidobacterium breve LMC 017: utilization as a functional starter culture for milk fermentation

This study was performed to discover bifidobacteria isolated from human intestines that optimally convert linoleic acid (LA) to conjugated linoleic acid (CLA) and to optimize the culture conditions of milk fermentation. One hundred and fifty neonatal bifidobacteria were screened for CLA-producing ability, and Bifidobacterium breve LMC 017 was selected as it showed about 90% conversion of free LA in MRS broth. The selected strain showed resistance at 0.5% LA in microaerophillic conditions. When monolinolein (LA 90%) was used as a substrate for CLA production, the conversion rate was lower compared to free LA, but the growth rate was unaffected during the milk fermentation. There was no significant difference in CLA production between aerobic and anaerobic conditions, and little decline in CLA was shown after the maximal CLA level had been reached. CLA production increased by 80% with 24 h of incubation in milk containing additional skim milk (5%), where the proteins may have facilitated the production of CLA by enhancing the interaction of substrate with the bacteria. CLA production did not decline after 9 h of fermentation and an additional 12 weeks of storage with other commercial starters. This demonstrates the possibility of using this strain as a costarter in the production of CLA-enriched yogurt.     

Gravinol ameliorates high-fructose-induced metabolic syndrome through regulation of lipid metabolism and proinflammatory state in rats

Using a rat model with fructose-induced metabolic syndrome, the effect of gravinol was investigated. Male Wistar rats were fed a 65% fructose diet and administered 10 or 20 mg of gravinol/kg of body weight/day for 2 weeks. High-level fructose feeding led to hyperglycemia, hyperlipidemia, hypertri-glyceridemia, and hypertension. On the other hand, the administration of gravinol significantly lowered serum glucose and total cholesterol levels. The tail arterial blood pressure was significantly elevated with the high-fructose diet. However, rats given gravinol showed a lower blood pressure as compared with fructose-fed control rats. In addition, the triglyceride (TG) levels in serum and lipoprotein fraction were dose-dependently reduced in rats fed gravinol. The decreases of hepatic TG and total cholesterol by gravinol were responsible for the down-regulation of hepatic sterol regulatory element binding protein (SREBP)-1. However, gravinol did not affect the protein levels of hepatic peroxisome proliferator-activated receptor-alpha and SREBP-2. Moreover, gravinol administration in the fructose-fed rats markedly reduced the glycosylated protein and thiobarbituric acid-reactive substance levels in the serum and hepatic mitochondria, and it inhibited the increase of the cyclooxygenase-2 protein level as a result of the down-regulation of nuclear factor kappa B (NF-kappaB). Furthermore, the decrease of anti-apoptotic bcl-2 protein levels and the increase of pro-apoptotic bax protein levels by the high-fructose diet were reversed by gravinol. These findings suggest that fructose-induced metabolic syndrome is attenuated by gravinol administration, which is associated with the reduction of serum lipids and protection against the proinflammatory state induced by oxidative stress.     

Growth-Inhibitory Effects of Pigmented Rice Bran Extracts and Three Red Bran Fractions against Human Cancer Cells: Relationships with Composition and Antioxidative Activities

We determined the phenolic, anthocyanin, and proanthocyanidin content of three brown, purple, and red rice brans isolated from different rice varieties using HPLC-PDA with the aid of 27 standards of known structure and matching unknown peaks to a spectral library of known compounds. Antioxidative capacities were determined by DPPH and ORAC and cell-inhibiting effects using an MTT assay. Based on the calculated IC(50) values, the light-brown bran had no effect, the purple bran exhibited a minor effect on leukemia and cervical cancer cells, and the red bran exhibited strong inhibitory effects on leukemia, cervical, and stomach cancer cells. High concentrations of protocatechuic acid and anthocyanins in purple bran and proanthocyanidins in red bran were identified. The red bran was further fractionated on a Sephadex column. Fraction 3 rich in proanthocyanidin oligomers and polymers had the greatest activity. Red bran has the potential to serve as a functional food supplement for human consumption.     

Physicochemical Properties and In Vitro Starch Digestibility of Cooked Rice from Commercially Available Cultivars in Canada

The influence of amylose content, cooking, and storage on starch structure, thermal behaviors, pasting properties, and rapidly digestible starch (RDS), slowly digestible starch (SDS), and resistant starch (RS) in different commercial rice cultivars was investigated. Long grain rice with high‐amylose content had a higher gelatinization temperature and a lower gelatinization enthalpy than the other rice cultivars with intermediate amylose content (Arborio and Calrose) and waxy type (glutinous). The intensity ratio of 1047/1022 cm^–1 determined by Fourier Transform Infrared (FT‐IR), which indicated the ordered structure in starch granules, was the highest in glutinous and the lowest in long grain. Results from Rapid ViscoAnalyser (RVA) showed that the rice cultivar with higher amylose content had lower peak viscosity and breakdown, but higher pasting temperature, setback, and final viscosity. The RDS content was 28.1, 38.6, 41.5, and 57.5% in long grain, Arborio, Calrose, and glutinous rice, respectively, which was inversely related to amylose content. However, the SDS and RS contents were positively correlated with amylose content. During storage of cooked rice, long grain showed a continuous increase in pasting viscosity, while glutinous exhibited the sharp cold‐water swelling peak. The retrogradation rate was greater in rice cultivars with high amylose content. The ratio of 1047/1022 cm^–1 was substantially decreased by cooking and then increased during storage of cooked rice due to the crystalline structure, newly formed by retrogradation. Storage of cooked rice decreased RDS content and increased SDS content in all rice cultivars. However, no increase in RS content during storage was observed. The enthalpy for retrogradation and the intensity ratio 1047/1022 cm^–1 during storage were correlated negatively with RDS and positively with SDS (P ≤ 0.01).     

Pterocarpan profiles for soybean leaves at different growth stages and investigation of their glycosidase inhibitions

Soybean leaves are eaten as seasonal edible greens in Korea. Analysis of the ethyl acetate extract of these leaves showed that it exhibited potent and selective neuraminidase inhibition, which began at the R3 stage and peaked at R7. Ten pterocarpans, including the new 6a-hydroxypterocarpan 10, were isolated from soybean leaves and their inhibition activities tested against a range of glycosidases. The relationship between structure and enzyme inhibition was investigated: 6a-hydroxypterocarpans exhibited much higher inhibition against neuraminidase (IC(50) = 2.4-89.4 μM) than α-glucosidase (IC(50) = 90.4-?>100 μM). Glyceollin VII (7) displayed 40-fold greater activity (IC(50) = 2.4 μM) against neuraminidase than α-glucosidase (IC(50) = 90.4 μM). On the other hand, coumestanes (1-3) were good α-glucosidase inhibitors (IC(50) = 6.0-42.6 μM). In kinetic analysis, the most potent neuraminidase inhibitors (5-10) were noncompetitive. HPLC analysis indicated that most pterocarpan synthesis began from the R3 stage, and a rapid change of pterocarpan concentrations was observed between the R4 and R7 stages.     

Identification and structure elucidation of a p-phenoxybenzaldehyde in bamboo shoots by HPLC-ESI/MS/MS and NMR

In this study, a derivative of p-phenoxybenzaldehyde in bamboo shoots was investigated. Bamboo shoots were ground and extracted with water, and an aqueous suspension was purified by SPE using Oasis HLB cartridges. After the SPE procedure, the analytes were analyzed by HPLC with refractive index detection (HPLC-RI). In the HPLC-RI analysis for sucralose, a putative sucralose was detected. In the subsequent HPLC-PDA analysis, the suspicious peak showed a unique UV absorption spectrum with the maximum wavelength at 285 nm indicating the existence of an aromatic ring. The contents of the unknown compound in bamboo shoot products ranged from 0.01 to 0.15 mg/g. The identity of the unknown compound was further confirmed by HPLC-ESI/MS/MS. The molecular weight of the unknown compound was determined to be 244. The chemical structure of the unknown compound was elucidated on the basis of NMR spectroscopic analyses ((1)H, (13)C, DEPT, COSY, HMQC, and HMBC). Finally, the structure of the unknown compound was characterized as 4-(4-dihydroxymethylphenoxy)benzaldehyde.     

Composition and mechanism of antitumor effects of Hericium erinaceus mushroom extracts in tumor-bearing mice

We investigated antitumor effects of the following four extracts of freeze-dried Hericium erinaceus mushrooms in Balb/c mice intracutaneously transplanted on the backs with CT-26 colon cancer cells: HWE, hot water extraction by boiling in water for 3 h; MWE, microwaving in 50% ethanol/water at 60 W for 3 min; and ACE and AKE, boiling in 1% HCl or 3% NaOH for 2 h. HWE and MWE with a higher content of β-glucans, determined by an assay kit, than ACE and MKE were active in all bioassays. Gas chromatography/mass spectrometry analyses showed the presence of 40, 27, 16, and 13 compounds, respectively, in the four extracts. Daily intraperitoneal (ip) injections of HWE and MWE for 2 weeks significantly reduced tumor weights by 38 and 41%. Tumor regressions were associated with changes in the following cancer biomarkers as compared to phosphate buffer (PBS)-treated control mice: 2.7- and 2.4-fold increases in cytolytic activity of splenic natural killer (NK) cells; restored nitric oxide production and phagocytosis in peritoneal macrophages to 95-98% of normal levels; ～2-fold increase in released pro-inflammatory cytokines tumor necrosis factor-α, interleukin-1β, and interleukin-6 from macrophages; and ～56 and ～60% reductions in the number of blood vessels inside the tumor. The pro-angiogenic factors vascular endothelial growth factor (VEGF), cyclooxygenase 2 (COX-2), and 5-lipoxygenase (5-LOX) were also significantly reduced in mRNA and protein expression by tumor genes. Enzyme-linked immunosorbent assay of tumor cells confirmed reduced expression of COX-2 and 5-LOX (32 and 31%). Reduced COX-2 and 5-LOX expression down-regulated VEGF expression, resulting in inhibition of neo-angiogenesis inside the tumors. The results indicate that induction of NK activity, activation of macrophages, and inhibition of angiogenesis all contribute to the mechanism of reduction of tumor size.     

Erythritol attenuates the diabetic oxidative stress through modulating glucose metabolism and lipid peroxidation in streptozotocin-induced diabetic rats

We investigated the effects of erythritol on rats with streptozotocin- (STZ-) induced diabetes mellitus. Oral administration of erythritol [100, 200, or 400 mg (kg body weight)(-1) day(-1) for 10 days] to rats with STZ-induced diabetes resulted in significant decreases in the glucose levels of serum, liver, and kidney. Erythritol also reduced the elevated serum 5-hydroxymethylfurfural level that is glycosylated with protein as an indicator of oxidative stress. In addition, thiobarbituric acid-reactive substance levels of serum and liver and kidney mitochondria were dose-dependently lower in the erythritol-treated groups than in the control diabetic group. Furthermore, the serum creatinine level was reduced by oral administration of erythritol in a dose-dependent manner. These results suggest that erythritol affects glucose metabolism and reduces lipid peroxidation, thereby improving the damage caused by oxidative stress involved in the pathogenesis of diabetes.     

Antioxidant activity of glyceollins derived from soybean elicited with Aspergillus sojae

The extract of soybean exposed to biotic elicitors such as food-grade fungus is known to have antioxidant activity. Glyceollins were major bioactive compounds present in soybean elicited by fungi and shown to have antifungal and anticancer activities. The purpose of present study was to evaluate the antioxidant activities of glyceollins by measuring ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, singlet oxygen quenching, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging, hydroxyl radical scavenging activity, and lipid peroxidation inhibition. In addition, the antioxidant potential of glyceollins were measured by a fluorescent probe, 2',7'-dichlorofluorescin diacetate (DCFDA), and dihydroethidium (DHE) in mouse hepatoma hepa1c1c7 cells in which they were insulted with H2O2 to generate reactive oxygen species (ROS). Glyceollins showed a strong reducing power and inhibited lipid peroxidation, with significant scavenging activities of radicals including singlet oxygen, superoxide anion, ABTS, and DPPH. We also found that glyceollins significantly suppressed H2O2-induced ROS production in hepa1c1c7 cells. Therefore, glyceollins deserve further study as natural antioxidants and nutraceuticals.     

Variation and correlation analysis of flavonoids and carotenoids in Korean pigmented rice (Oryza sativa L.) cultivars

Flavonoids and carotenoids of pigmented rice ( Oryza sativa L.), including five black cultivars and two red cultivars, from Korea were characterized to determine the diversity among the phytochemicals and to analyze the relationships among their contents. Black cultivars were higher in flavonoids and carotenoids than the red and white cultivars. The profiles of eight phytochemicals identified from the rice grains were subjected to principal component analysis (PCA) to evaluate the differences among cultivars. PCA could fully distinguish between these cultivars. The Heugjinjubyeo (BR-1) and Heugseolbyeo (BR-2) cultivars were separated from the others based on flavonoid and carotenoid concentrations. Flavonoid contents had a positive correlation with carotenoid contents among all rice grains. The BR-1 and BR-2 cultivars appear to be good candidates for future breeding programs because they have simultaneously high flavonoid and carotenoid contents.