• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.51-56
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• 2002

An antitumor substance was purified from the culture filtrate of phytopathogenic fungus Alternaria brassicicola SW-3 isolated from soil of a chinese cabbage patch, and its characteristics were investigated. Antitumor activity of A. brassicicola SW-3 was measured by MTT assay. The cytotoxic activity against human cancer cell line was detected in the culture filtrate of A. brassicicola SW-3, but no activity found in mycelium. Antitumor substance was isolated from the culture broth by ethyl acetate extraction and purified by silica gel column chromatography. Structure of the purified compound was analyzed by the instrumental analysis such as $^1$H-NMR, $^{13}$ C-NMR and IR spectroscopy. The purified fungal metabolite of an A. brassicicola SW-3, consists of 11 carbon chain with two hydroxyl groups and two epoxides which is identical to depudecin. The $IC_{50}$/ values of the active compound identified as depudecin were $69\mu$g/mL and $57\mu$g/mL against mouse melanoma B16BL6 cell line, and human hepatoma SK-HEP1 cell line, respectively.

• YAKHAK HOEJI
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• v.39 no.6
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• pp.661-665
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• 1995

Ginsenoside-Rg$_{1}$(G-Rg$_{1}$) has been shown to stimulate DNA synthetic activity in SK-HEP-1 cells. This study was therefore designed to determine in SK-HEP-1 cells whether the stimulatory effect of G-Rg$_{1}$ may be mediated by protein kinase C (PKC) which is known to play a key role in the signal transduction pathway leading to the cell proliferation. Using the tn situ PKC assay method, the PKC enzyme activity was determined in SK-HEP-1 cell cultures in response to G-Rg$_{1}$ at 3*10$^{-5}$ M or phorbol 12-myristate 13-acetate(PMA) at 10$^{-6}$ M which in the enzyme activity by 1.5- and 7-fold, respectively. Furthermore, G-Rg$_{1}$, was also able to synergistically increase the enzyme activity by 11-fold m the cell cultures in the presence of PMA. These stimulatory effects of G-Rg$_{1}$ or PMA on the DNA synthetic activity and the PKC activity were ablished by a specific PKC inhibitor, GF109203X. These results suggest that the stimulatory effect of G-Rg$_{1}$ on the DNA synthetic activity may be partly due to stimulation of PKC-mediated signal transduction pathway leading to the proliferation of SK-HEP-1 cells.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.184-189
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• 2004

Rice wine cake (RWC) is the solid waste obtained after rice wine fermentation. For the mass production of the spores of yeast Saccharomyces from RWC, the optimum pretreatment condition of RWC, the optimum composition of culture medium, and the optimum culture condition were examined. For sporulation, yeast cells were grown in the pre sporulation medium (PSM), transferred into sporulation medium (SM) containing 1 % potassium acetate, and incubated in a rotary shaking incubator at $25^{\circ}C$ for 4 days. The supernatant of the mixture of RWC and water was used as the presporulation medium (PSM). The optimum temperature and time for the pre-incubation of the mixture of RWC and water (1:2) to obtain maximum sporulation yield were $V^{\circ}C$ and 24 hr, respectively, and optimum culture time in PSM was 48 hr. Using these optimum conditions, the asci number obtained was 0.72$ 1.06${\times}$10^{8}$$m\ell$. The addition of wheat coat koji into SM increased the final number of asci to beTEX>$10^{8}$ $m\ell$. Spores were formed in the SM with the initial pH of 7-11, but no spores were formed in the SM with the initial pH of 5. To save the time and effort to pretreat the RWC, 2% and 0.5% RWC without any pretreatment were directly added into PSM containing 1 % brown sugar and SM, respectively, and the maximum asci number of $1.27${\times}$10^{8}$ /$m\ell$ was obtained.

• Microbiology and Biotechnology Letters
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• v.22 no.5
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• pp.507-514
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• 1994

Acetyl xylan esterase was produced by E. coli HB101 harboring a recombinant plasmid pKMG6 which contained the estI gene of Bacillus stearothermophilus. The maximum production was observed when the E. coli strain was grown at 37$\circC for 12 hours in the medium containing 0.5% acetyl xylan, 1.0% tryptons, 1.0% sodium chloride, and 0.5% yeast extract. The esterase produced was purified to homogeneity using a combination of ammonium sulfate fractionation, DEAE Sepharose CL-6B ion exchange chromatography and Sephacryl S-200 gel filtration. The native enzyme had an apparent molecular mass of 60 kd and was composed of two identical subunits of 29 kd. The N-terminal amino acid sequence of the polypeptide was Ala-X-Leu-Gln- Ile-Gln-Phe-X-X-Gln. The acetyl esterase displayed a pH optimum of 6.5 and a temperature opti- mum of 45$\circC. The heavy metal ions such as Ag$^{++}$, Hg$^{++}$ and Cu$^{++}$ inhibited nearly completely the activity of the esterase, and no specific metal ion was found to be required for the enzyme activity. The enzyme readily cleaved MAS, $\beta$-D-glucose pentaacetate, $\alpha$-naphthyl acetate, $\rho$-nitrophenyl acetate as well as acetyl xylan, but had no activity on $\rho$-nitrophenyl propionate, $\beta$-nitrophenyl butyrate or $\beta$-nitrophenyl valerate. The Km and Vmax values for MAS were 2.87 mM and 11.55 $\mu$mole/min, respectively. Synergistic behavior was demonstrated with a combination of xylanase and esterase from B. stearothermophilus in hydrolyzing acetyl xylan.

• The Korea Journal of Herbology
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• v.30 no.2
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• pp.31-36
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• 2015

Objectives : In this study, we demonstrated the antioxidant activities and the inhibitory effect on oxidative DNA damage of ethyl acetate fractions extracted from Sophorae Flos and Sophorae fructus. Methods : Sophorae Flos and Sophorae fructus were extracted with methanol(MeOH) and divided to Petroleum ether, Ethyl acetate(EtOAC) and Water fraction. The antioxidant activities were conducted by the 1,1-diphenyl-2-picryl hydrazyl(DPPH) radical, 2, 2'-Azine-bis(3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt(ABTS) radical scavenging assay, $Fe^{2+}$ chelating assay and Reducing power assay. The inhibitory effect of DNA damage were characterized on ${\varphi}$ X-174 RF I plasmid DNA cleavage assay. In addition, we analyzed the Total phenol contents and the Vitamin C contents of Sophorae Flos and Sophorae fructus. Results : The results of DPPH were 92.71% and 94.72%, ABTS were 87.16% and 62.44%, and $Fe^{2+}$ chelating were 95.81% and 85.11% at $200{\mu}g/m{\ell}$ of Sophorae Flos and Sophorae fructus respectively. The Sophorae Flos showed stronger effect than Sophorae fructus in Reducing Power assay. Total phenol content was 111.77 mg/g and 122.54 mg/g, and Vitamin C content was 2.59 mg/g and 3.03 mg/g. Also both Sophorae Flos and Sophorae fructus have inhibitory antioxidant effect on ${\varphi}$ X-174 RF I plasmid DNA cleavage assay. Conclusions : Over all, this study suggests that Sophorae Flos and Sophorae fructus can be used as not only effective antioxidant but also natural medicine.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.11
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• pp.1583-1591
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• 2015

The present study was conducted to determine the effect of lipid sources with different fatty acid profiles on nutrient digestion and ruminal fermentation. Ten rumen and duodenal fistulated Nellore steers (268 body weight${\pm}27kg$) were distributed in a duplicated $5{\times}5$ Latin square. Dietary treatments were as follows: without fat (WF), palm oil (PO), linseed oil (LO), protected fat (PF; Lactoplus), and whole soybeans (WS). The roughage feed was corn silage (600 g/kg on a dry matter [DM] basis) plus concentrate (400 g/kg on a DM basis). The higher intake of DM and organic matter (OM) (p<0.001) was found in animals on the diet with PF and WF (around 4.38 and 4.20 kg/d, respectively). Treatments with PO and LO decreased by around 10% the total digestibility of DM and OM (p<0.05). The addition of LO decreased by around 22.3% the neutral detergent fiber digestibility (p = 0.047) compared with other diets. The higher microbial protein synthesis was found in animals on the diet with LO and WS (33 g N/kg OM apparently digested in the rumen; p = 0.040). The highest C18:0 and linolenic acid intakes occurred in animals fed LO (p<0.001), and the highest intake of oleic (p = 0.002) and C16 acids (p = 0.022) occurred with the diets with LO and PF. Diet with PF decreased biohydrogenation extent (p = 0.05) of C18:1 n9,c, C18:2 n6,c, and total unsaturated fatty acids (UFA; around 20%, 7%, and 13%, respectively). The diet with PF and WF increased the concentration of $NH_3-N$ (p<0.001); however, the diet did not change volatile fatty acids (p>0.05), such as the molar percentage of acetate, propionate, butyrate and the acetate:propionate ratio. Treatments PO, LO and with WS decreased by around 50% the concentration of protozoa (p<0.001). Diets with some type of protection (PF and WS) decreased the effects of lipid on ruminal fermentation and presented similar outflow of benefit UFA as LO.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.7
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• pp.962-970
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• 2012

This study aimed to investigate the effects of ruminal infusion of garlic oil (GO) on fermentation dynamics, fatty acid (FA) profile, and abundance of bacteria involved in biohydrogenation in the rumen. Six wethers fitted with ruminal fistula were assigned to two groups for cross-over design with a 14-d interval. Each 30-d experimental period consisted of a 27-d adaptation and a 3-d sample collection. Goats were fed a basal diet without (control) or with GO ruminal infusion (0.8 g/d). Ruminal contents collected before (0 h) and at 2, 4, 6, 8, and 10 h after morning feeding were used for fermentation analysis, and 0 h samples were further used for FA determination and DNA extraction. Garlic oil had no influence on dry matter intakes of concentrate and hay. During ruminal fermentation, GO had no effects on total VFA concentration and individual VFA molar proportions, whereas GO increased the concentrations of ammonia nitrogen and microbial crude protein (p<0.05). Compared with control, GO group took a longer time for total VFA concentration and propionate molar proportion to reach their respective maxima after morning feeding. The ratio of acetate to propionate in control reduced sharply after morning feeding, whereas it remained relatively stable in GO group. Fatty acid analysis showed that GO reduced saturated FA proportion (p<0.05), while increasing the proportions of C18, t11-18:1 (TVA), c9,t11-conjugated linoleic acid (c9,t11-CLA), t10,c12-CLA, and polyunsaturated FA (p<0.05). The values of TVA/(c9,t11-CLA+TVA) and C18:0/(TVA+C18:0) were reduced by GO (p<0.05). Real-time PCR showed that GO tended to reduce Butyrivibrio proteoclasticus abundance (p = 0.058), whereas GO had no effect on total abundance of the Butyrivibrio group bacteria. A low correlation was found between B. proteoclasticus abundance and C18:0/(TVA+C18:0) (p = 0.910). The changes of fermentation over time suggested a role of GO in delaying the fermentation process and maintaining a relatively modest change of ruminal environment. The inhibitory effects of GO on the final step of biohydrogenation may be related to its antibacterial activity against B. proteoclasticus and other unknown bacteria involved.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.3
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• pp.619-630
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• 2009

To investigate the hair growth activity of fractions and extract of Arisaematis Rhizoma in the hair removed skin of normal and spontaneous alopecia areata model in C57B/6N mice. These experiments were performed with the macroscopic, microscopic, immunohistochemical(VEGF, c-kit, PKC-${\alpha}$, TGF and FGF) and RT-PCR(TGF-${\beta}$, IGF, prolactin and placenta lactogen) methods. The results were as follows: Macroscopic observation after topical application of vehicle, 50% EtOH as control and extract of Arisaematis Rhizoma to the hair removed skin of C57BL/6N mice on the 9th, 11th and 15th day. Extensive hair growth activity was observed in treated group with extract of Arisaematis Rhizoma on the 9th, 11th and 15th day. In Arisaematis Rhizoma extracts treated group, hair follicles of middle stage of anagen was observed and it were grown down to subcutaneous tissue of skin in all the normal mice on 15th day. But in control group, most of hair follicles of telogen phase was observed in skin. The treatment of extract of Arisaematis Rhizoma increased expression of IGF(145%) and placenta lactogen(108%) in the skin of normal C57BL/6N mice on the 11th day compared to control group(100%). But expression of TGF-${\beta}$(90%) and prolactin(91%) decreased in the skin of normal C57B/6N mice on the 11th day compared to control group(100%). After application of fractions(chloroform, ethyl acetate and water fractions) of Arisaematis Rhizoma extract for 9th day, hair growth effect was observed in whole skin area in 50% of normal mice. But in control group, hair growth effect was not observed in whole skin area of normal mice. Immunoreactive density of VEGF, c-kit, PKC-${\alpha$ and FGF in skin of fractions of Arisaematis Rhizoma extracts was strongly stained in epidermis, bulge, secondary hair germ cells, cutaneous trunci m., subcutaneous tissue, root sheath compare to control group on the 9th day. In spontaneous alopecia areata model, The hair growth activity of Arisaematis Rhizoma extrat treated group(75%) was observed to be strong compared to control group(O%) on 7th day. These experiments suggest that fractions and extracts of Arisaematis Rhizoma may stimulate the topical hair growth activity. Thus it can be useful for treatment of alopecia areata.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.1
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• pp.115-118
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• 2003

This study was conducted to determine optimal extraction condition and free radical scavenging effect of ethanol extracts of Campell Early grape seeds and skins at different temperatures, and of organic solvent fractionated from ethanol extracts of grape seeds and skins. The free radical scavenging effect was tested using 1,1-diphenyl-2-picrylhydrazyl (DPPH) method and expressed as dose required for a 50% reduction of DPPH radical (RC$_{50}$). The extraction yields were increased with increasing the extraction temperature but it was not related with ethanol concentrations. The ethanol extract of grape seeds obtained at 5$0^{\circ}C$ showed the more stronger RC$_{50}$ (16.8 $\mu\textrm{g}$/mL) than those of both 3$0^{\circ}C$ and 78$^{\circ}C$. The ethylacetate fraction obtained from ethanol extract of grape seed showed the strongest RC$_{50}$ (15.4 $\mu\textrm{g}$/mL). Overall, both ethanol extracts and their fractions of grape seeds showed significantly stronger free radical scavenging effect than those of skin extracts.racts.

• Journal of Applied Biological Chemistry
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• v.55 no.3
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• pp.169-172
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• 2012

The aim of this research was to investigate the antibacterial activity of Clerodendron trichotomum. Antibacterial activities of the n-hexane, methylene chloride (MC), ethyl acetate, and n-butanol fractions from C. trichotomum were tested against Staphylococcus aureus, Escherichia coli, and Helicobacter pylori. The n-hexane and MC fractions showed antibacterial activity against H. pylori at a concentration of 1.7 mg/mL and showed inhibition zones of 10 and 11 mm in disc assay, respectively. Further testing of 22-dehydroclerosterol and ${\beta}$-amyrin (each 3.4 mg/mL) from the MC fraction of C. trichotomum revealed moderate antibacterial effects against E. coli, S. aureus, and H. pylori. In particular, ${\beta}$-amyrin showed clear zones of 12 and 13 mm against E. coli and H. pylori, respectively, suggesting its potential as an antibacterial agent. The active compounds from C. trichotomum might provide a promising therapeutic agent against infections by E. coli, S. aureus, and H. pylori.