This study was carried out to evaluate the antimicrobial effect of red ginseng (Panax ginseng C.A. Meyer) against several foodborne pathogens including Staphylococcus aureus, Escherichia coli, Candida albicans and Aspergillus niger. The antimicrobial effect was determined by agar diffusion method using red ginseng extract, crude saponin and non-water-soluble fractions. Red ginseng extract showed antimicrobial effect against S. aureus, but not C. albicans or A. niger. The extract showed anti-bacterial activity at concentration above 30% against S. aureus, which cause both food poisoning and atophic dermatitis. Crude saponin showed antibacterial activity above 7.5% against the bacterium. However, the ginsenosides purified from crude saponin showed no antimicrobial activities at 100-200 ㎍/mL. To investigate the mode of growth inhibition, red ginseng extract and crude saponin were added to 0.85% NaCl solution containing S. aureus and then incubated at 35℃ for 12 h. The results showed that viable cells were rapidly reduced in above 10% concentration of red ginseng extract and above 2% of crude saponin, respectively. However, the crude saponin and red ginseng extract did not inhibit the bacterial cells completely at those same concentrations. On the other hand, whereas all non-water-soluble fractions showed inhibition zones above 10 mm against S. aureus, they showed no inhibition effects against E. coli, C. albicans or A. niger. The methanol fraction-1 (MF-1) showed the highest antibacterial activity against S. aureus, and the MIC (minimal inhibitory concentration) was 0.625 mg/mL. These results suggest that red ginseng extract, crude saponin and non-water-soluble fractions show selective antibacterial activity against S. aureus, and non-water-soluble fractions might be used as natural antibacterial agents.
The purpose or this study was to investigate the effect or H $_2$O fraction or Dioscorea japonica Thunb(DJT) and selenium(Se) on the lipid peroxidation in streptozotocin (STZ)-induced diabetic rats. Male Sprague-Dawley rats weighing 180∼220g were divided into 5 groups: One normal rat group and 4 diabetic rat groups(the STZ-Control group, the DJT group, the DJT-Se group and the Se group). Diabetes was induced in the male rats by injection of STZ into tail vein at a dose of 45 mg/kg. The H$_2$O-fraction of DJT(500 mg/kg) was administered orally for 14 days. The supplementation was achieved with the AIN-76 recommendation diet by adding 2 mg/kg diet of selenium as Na$_2$SeO$_3$ which was prepared freshly everyday. The levels of glycogen in liver and muscle and protein in kidney, liver and muscle were measured. The liver concentrations of cholesterol and triglyceride were analyzed. Also, the malondialdehyde(MDA) levels in kidney, liver and lung were determined. The glycogen levels of liver and muscle in diabetic groups were not significantly different from the normal group. The protein concentrations of kidney, liver and muscle were not significantly different either, but the level of muscle protein was higher than STZ-Control group. The levels of liver cholesterol were significantly different between normal and STZ-Control groups and decreased in all diabetic experimental groups fed on H$_2$O-fraction of DJT and Se supplementation compared with the STZ-Control group. The levels of liver triglyceride were higher in the DJT-Se group than the STZ-Control group. The concentrations of MDA in lung decreased greatly by the administration of Se among all and the concentration of liver MDA was significantly reduced and that of DJT-Se group was the lowest. In conclusion, the results indicated that the administration of H$_2$O-fraction of DJT with selenium supplementation has a synergistic antioxidative effect by influencing on lipid metabolites and peroxidation especially in liver.
This study was carried out to evaluate the antiobesity effects of Crataegus fructus in 3T3-L1 adipocytes and mice fed a high fat diet (high fat 45% cal). The inhibitory effect of methanol extract from Crataegus fructus on lipid accumulation in 3T3-L1 adipocytes was quantified using Oil red O staining. Compared with the control, lipid accumulation was significantly decreased by 10-25% with treatment with Crataegus fructus extract at a concentration of 600-2,000 ug/ml. Three-week old ICR mice (n=24) were randomly divided into four groups (T0: normal diet, T1: high fat diet, T2: high fat diet and 50 ug of Crataegus fructus extract, T3: high fat diet and 100 ug of Crataegus fructus extract) and were fed an experimental diet for 5 weeks. At the end of the experiment, body weight gain in the T1 group (3.9${\pm}$0.24 g) was higher than that in the T0 group (2.56${\pm}$0.14 g), while body weight gain in the T2 (3.02${\pm}$0.25 g) and T3 (2.58${\pm}$0.16 g) groups was significantly reduced as compared with that of the T1 group. Moreover, liver weight in the T1 (4.8${\pm}$0.17 g) and T2 (4.8${\pm}$0.16 g) groups was significantly higher than that of the T0 (4.05${\pm}$0.16 g) and T3 (4.57${\pm}$0.10 g) groups, while kidney weight was significantly lower than that of the T0 and T3 groups (p<0.05). The levels of total cholesterol and triglyceride in serum in the T2 and T3 groups were significantly decreased compared to the T1 group. These results suggest that Crataegus fructus can be used as functional materials in food and medicine.
Recently, use of natural medicine is getting more attention, and some of them are believed to be effective in the treatment of periodontitis. Among them, the seeds of safflower(Carthamus tinctrorius L.) have been proven to be effective through its use in bone diseases such as fracture and osteoporosis. During the last few years, studies using the seeds of safflower gown in Korea have been active, and it has been reported that safflower seed extract increase the proliferation and the alkaline phosphatase(ALP) activity of human periodontal ligament fibroblast(hPDLF), osteoblast, and that they promote the mineralization process. In animal studies, when safflower seed extract were administered orally new bone formation was promoted. Recently, in an effort to find out the most effective osteogenic components, among many components of the safflower seed, various safflower seed fraction extracts were obtained by multistep extraction of the safflower components using various solvents. Among these, saf-M-W fraction extracted by methanol and water was most effective in increasing osteogenic potential of osteoblasts. In this study, the effect of safflower seed fraction extract, saf-M-W, on the growth and differentiation of hPDLF and MC3T3-E1 cell was investigated. The toxicity of saf-M-W on both cells was measured using M'IT(3-(4,5dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide) test, and ALP activity was measured using the colorimetric assay of hPDLF. In addition, in MC3T3-El cells, the expression of ALP, bone sialoprotein(BSP) mRNA was observed using Northern blot, and the mineralized nodule formation Was observed using von Kossa stain and phase-contrast microscope. 1. In concentrations below $10{\mu}g/ml$, saf-M-W didn't show any toxicity on hPDLF and MC3T3-El cell. 2. The change in saf-M-W concentration had no effect on the ALP activity of hPDLF. 3. In MC3T-E1 cells, mRNA expressions of ALP and BSP were greater in the experimental group treated with $10{\mu}g/ml$ concentration of saf-M-W compared with the control group. 4. In MC3T3-El cells, abundance of mineralized nodules were formed in the experimental group treated with $10{\mu}g/ml$ Concentration of saf-M-W, while no mineralized nodule was formed in the control group. These results suggest that safflower seed fraction extract, saf-M-W. didn't show any toxicity on hPDLF and MC3T3-E1 cell at concentrations below $10{\mu}g/ml$ and effectively enhanced the differentiation and osteogenic potential of MC3T3-El cell.
The indigenous Saccharomyces cerevisiae strains S13 and D8 were isolated at the microbial succession stage during spontaneous fermentation of Campbell Early wine as a resistant to potassium metabisulfite and a high sugar concentration. In this study, the fermentation characteristics of Campbell Early wine were investigated and compared with those of S. cerevisiae W-3, an industrial wine yeast. Alcohol production by the two strains was delayed at the initial fermentation stage, but increased fast when the fermentation continued. After the fermentation, the S13 and D8 wines contained 12.6% and 13.2% (v/v) alcohol, respectively, which were significantly higher than the alcohol content of the W-3 wine (12%, v/v). No marked differences were observed in the residual soluble solid content and the pH. However, the S13 and D8 wines showed high levels of total acid content, including malic and lactic acids. Especially, the lactic acid content was 8.9-fold in the S13 wine and six-fold in the D8 wine, compared with that of the W-3 wine. The two strains produced a higher level of acetaldehyde and a lower amount of methanol in the wine than the W-3 strain. The iso-Butanol content was lower in the two indigenous yeast wines with similar levels of n-propanol and iso-amyl alcohol contents than that in the W-3 wine. In the sensory evaluation, the S13 and D8 wines had higher scores for their color, flavor, taste and overall preferences than the W-3 wine. Especially, the S13 and D8 wines had much higher scores than the W-3 wine for flavor and color, respectively.
Fermentation characteristics of chestnut-added yakju prepared using various proportions of raw materials such as rice koji, rice flour, cornflour koji and cornflour were investigated. The pH of chestnut-added yakju prepared with cornflour koji and saccharified cornflour showed a higher value than that of chestnut-yakju prepared with rice koji and saccharified rice flour. The total acid content of chestnut-added yakju prepared with rice koji and saccharified rice flour was higher than that of chestnut-added yakju prepared using cornflour koji and sacharified cornflour. The reducing of sugar in chestnut-added yakju prepared with saccharified rice flour or saccharified cornflour was rapid at the first brewing stage, decreased dramatically after 2 days, and then decreased slowly after 5 days of fermentation. The value of L and a, the Hunter values, were high in chestnut-added yakju prepared with cornflour koji, and value b was high in chestnut-added yakju with rice koji. The content of iso-amyl alcohol was the highest of seven kinds of fusel oil found in chestnut-added yakju. Ethanol content increased to $17.6{\sim}18.2%$(v/v) after 8 days of fermentation. The content of lactic acid was the highest of all organic acids in the chestnut-added yakju. Sensory test results on chestnut-added yakju prepared with saccharifed corn flour showed that if rice flour is used as a sugar supplement for chestnut, the yakju prepared using koji had better flavor and taste. If cornflour was used in the preparation of chestnut-added yakju, the used of saccharified cornflour offered superior flavor and taste.
The anti-oxidant properties of $Lactuca$$indica$ were determined using in-vitro assay systems. The vitamin C contents of the leaf and root extracts were 24.14 and 0.38 mg/100 g, respectively. The total polyphenol contents of the leaf and root extracts were 42.8 and 7.66 mg/g, and their flavonoid contents were 23.09 and 0.77 mg/g. The leaf extract showed higher DPPH and ABTS radical scavenging ability than the root extract at all the extract concentrations. Especially, the ABTS radical scavenging ability of the leaf extract was 92.3% at a concentration of 5 mg/mL. The reducing power was increased with the increase in the concentration of extracts, and the leaf extract had a higher reducing power than the root. The $Fe^{2+}$-chelating ability of the leaf and root were 97.2% and 34.3% at 14 mg/mL, respectively. The $IC_{50}$ values of the leaf for DPPH, its ABTS radical scavenging ability, and its $Fe^{2+}$-chelating ability were 0.19, 2.7, and 6.27 mg/mL, respectively, and the leaf extract showed lower $IC_{50}$ values than root extract. These results show that the $L.$$indica$ leaf extract contained high amounts of anti-oxidative compounds and had higher anti-oxidant activity levels than the root extract. It is suggested that Lactuca indica is very high in availability as a functional food and in its materials.
Purpose: This study was performed to investigate the effects of immune cell activation and the antitumor effect for the combination of treatment with X-irradiation and E/eutherococcus senticosus Maxim Root (ESMR) on mouse tumor cells. Materials and Methods: ESMR (250g) was extracted with 80% methanol, concentrated under decompression and lyophilized. To determine whether ESMR is able to activate the immune cells or not, the proliferation of splenocytes in vitro and the number of B cells and T cells in splenic lymphocytes in ESMR-pretreated mice were evaluated. X-irradiation was given to the mouse fibrosarcoma tumor cells (FSa II) by 250 kv X-irradiation machine. The cytotoxicity of ESMR was evaluated from its ability to reduce the clonogenecity of FSa II cells. In X-irradiation alone group, each 2, 4, 6 and 8 Gy was given to FSa II cells. In X-irradiation with ESMR group, 0.2 mg/ml of ESMR was exposed to FSa II cells for 1 hour before X-irradiation. Results: The proliferation of cultured mouse splenocytes and thymocytes were enhanced by the addition of ESMR in vitro. The number of B cells and T cells in mouse splenic lymphocytes was significantly increased in ESMR pretreated mice in vivo. In FSa II cells that received a combination of 0.2 mg/ml of ESMR with X-irradiation exposure, the survival fraction with a dose of 2, 4 and 6 Gy was $0.39{\pm}0.005$, $0.22{\pm}0.005$ and $0.06{\pm}0.007$, respectively. For FSa II cells treated with X-irradiation alone, the survival fraction with a dose of 2, 4 and 6 Gy was $0.76{\pm}0.02$, $0.47{\pm}0.008$ and $0.37{\pm}0.01$. The difference in the survival fraction of the mouse FSa II cells treated with and without ESMR was statistically significant (p<0.05). Conclusion: Treatment with ESMR increased cell viability of mouse splenocytes in vitro and especially the subpopulation of B cells and T cells in splenocytes in ESMR-pretreated mice. However, treatment with ESMR did not increase the level of Th and Tc subpopulations in the thymocytes. Treatment with the combination of ESMR and X-irradiation was more cytotoxic to mouse tumor cells than treatment with X-irradiation alone; this finding was statistically significant.
Sorafenib is a multikinase inhibitor and an oral anticancer drug approved for the treatment of patients with advanced renal cell carcinoma and those with unresectable hepatocellular carcinoma. The purpose of this study was to develop an efficient method of the determination of sorafenib in human plasma using tandem mass spectrometry coupled with liquid chromatography (LC/MS/MS) and validate the method by the guidelines of the Korean Food and Drug Administration (KFDA). Plasma samples ($100{\mu}l$) were added with chlorantraniliprole as an internal standard and then mixed with the 0.1% formic acid-containing extraction solution composed of isopropyl alcohol and ethyl acetate (1:4, v/v). After centrifugation, the supernatant was concentrated at $45^{\circ}C$ under negative pressure and centrifugal force. The residue was reconstituted with a mobile phase and injected into the HPLC instrument using a reverse phase Waters XTerra$^{TM}$ C18 column (particle size $3.5{\mu}m$). Liquid chromatography was carried out within the run time of 5 min using a mobile phase composed of buffer (0.1% formic acid and 10 mM ammonium formate), methanol, and acetonitrile (1:6:3, v/v/v). The analytes were monitored by tandem mass spectrometry in the multiple reaction monitoring method programmed to detect sorafenib at 'm/z 465.2 ${\rightarrow}$ 252.5' and chlorantraniliprole at 'm/z 484.4 ${\rightarrow}$ 286.2' with positive electrospray ionization mode ($ES^+$). The result showed the proper linearity ($r^2$ > 0.99) over the range of 2,000-5,000 ng/ml with good accuracy (90.7-103.9%) and precision (less than 10%). The newly developed method using LC/MS/MS was validated by the guideline of KFDA and identified as more sensitive compared to the previous methods.
A functional screen of 60,672 fosmid metagenomic clones amplified from marine sediment obtained from the Dokdo islets in Korea identified the gene EstES1, whose product, EstES1, displayed lipolytic properties on tributyrin-supplemented media. EstES1 is a 576 amino acid protein with a predicted molecular weight of 59.4 kDa including 37 N-terminal leader amino acids. EstES1 exhibited the highest sequence similarity (44%) to a carboxylesterase found in Haliangium ochraceum DSM14365. Phylogenetic analysis indicated that EstES1 belongs to a currently uncharacterized family of lipases. Within the conserved domain, EstES1 retains the catalytic triad that consists of the consensus penta-peptide motif, GESAG. EstES1 demonstrated a broad substrate specificity toward the long acyl group of ethyl esters (C2-C12), and its optimal activity was recorded toward p-Nitrophenyl butyrate (C4) at pH 9.0 and $40^{\circ}C$ (specific activity of 255.4 U/mg). The enzyme remained stable in the ranges of $60-65^{\circ}C$ and pH 9.0-10.5 and in the presence of methanol, ethanol, isopropanol, and dimethyl sulfoxide. Therefore, EstES1 has potential for use in industrial applications involving high temperature, organic solvents, and/or alkaline conditions.
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