• The Journal of Korean Medicine
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• v.26 no.1 s.61
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• pp.37-45
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• 2005

In the present study, we examined the effect of crude methanolic extract (CME) from Euonymus alatus (Thunb.) Sieb on arachidonic acid (AA) cascade in SKBR3 human breast cancer cell line. CME had a potent inhibitory activity of prostaglandin E2 (PGE2) release induced by A23187, a $Ca^{2+}$ ionophore. The inhibition was concentration-dependent, with the 50 value of about 5 M. CME had no inhibitory effect on A23187-induced phosphorylation of p42/p44 extracellular signal regulated kinase/mitogen-activated protein kinase or on the liberation of [14C]-AA from the cells labeled with [14C]-AA. However, CME concentration-dependently inhibited the conversion of AA to $PGE_2$ in microsomal preparations, showing its possible inhibition of cyclooxygenase (COX). In enzyme assay in vitro, CME inhibited the activities of both constitutive COX (COX­I) and inducible COX (COX-2) in a concentration-dependent manner, with the 50 values of about 0.8 and 2M, respectively. Lineweaver-Burk plot analysis indicated that CME competitively inhibited the activities of both COX-l and -2. This study is a first demonstration that CME directly inhibits COX activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.15 no.1
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• pp.22-26
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• 1986

This investigation was carried out to study the effect of heating treatment on the saponin fractions in ginseng extract and crude saponin. The changes of saponin patterns and amounts were investigated using HPLC and compared with peak area of each fraction. Shape of crude saponin was changed more easy to compare with in ginseng extract. The more extracting temperature risen and treated time longer, the more changes of saponin amounts and patterns were shown, expecially at $100^{\circ}C$ over. All of the saponin fraction except ginsenoside-Rd were relatively unstable in heating treatment. Suitable extracting condition was extracted at $80^{\circ}C$ for 40 hours with $H_2O$.

• The Journal of Natural Sciences
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• v.4
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• pp.69-83
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• 1991

To determine optimum ethanol concentration and clarification time in ethanol clarification of red ginseng extract(RG-Ext), physical properties, crude saponin recovery of clarified RG-Ext. and stability of red ginseng drink prepared from clarified RG-Ext. were investigated. Color intensity, redness(a value), viscosity and yield of clarified RG-Ext. were decreased in proportion to the increase of ethanol concentration and clarification time, but transmittance, brightness(L value) and yellowness (b value) were decreased. Crude saponin recovery of clarified RG-Ext. were not change significantly by the increase of ethanol concentration. Red ginseng drink prepared from 50-90% ethanol clarified RG-Ext. were stable without precipitation until six months at the storage of $0-5^\circC$ and $40^\circC$.

• Korean Journal of Microbiology
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• v.23 no.4
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• pp.259-264
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• 1985

The research was carried out for the purpose of finding effects of gerbal saponins on aflatoxin synthesis by Aspergillus parasitics NRRL 2999. A. parasiticus with $10^6$ conidia were grown at $30^{\circ}C$ for 9 days on the enriched medium that is optimum for the frowth and aflatoxins production by the mold. The inhibitory effect on the growth and aflatoxins produced by the mold occurred in the presence of 0.36% of crude red-ginseng saponin showing both the growth and aflatoxins production come to 62.3% (growth), 38.7% (aflatoxin $B_1$) and 22.9% (aflatoxin $G_1$) of the control. Thd next effective saponin to inhibit the growth and aflatoxins production was from burdock seeds. However, saponin extracted from honeysuckle flowers had no inhibitory effect. The mold caused no changes in the pH of the medium when it contained red-ginseng saponin. Red-ginseng saponin was more effective than the white-ginseng in inhibiting both the growth and aflatoxin production.

• Parasites, Hosts and Diseases
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• v.38 no.3
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• pp.159-166
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• 2000

The present study intended to verify activities of cysteine proteinase of Pneumocystis carinii from rats and to purify the enzyme. In order to exclude the contamination of host-derived enzymes, concentrates of P. carinii was primarily treated with a mixture of proteinase inhibitors before Iysis of P carinii. A 68-kDa cysteine proteinase was finally purified from the crude extract of P. carinii by 4 sequential chromatographic methods. The enzyme showed an optimal activity at pH 5.5 in 0.1 M sodium acetate, and its activity was specifically inhibited by L-trans-epoxy-succinylleucylamido (4-guanidino) butane (E-64) and iodoacetic acid, suggesting that the enzyme is a cysteine proteinase. The 68-kDa proteinase weakly digested rnacrornolecules such as collagen, hemoglobin and fibronectin. The present study demonstrated the activity of cysteine proteinase at the 68-kDa band of P. carinii, and purified and characterized the molecule.

• Preventive Nutrition and Food Science
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• v.1 no.1
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• pp.111-116
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• 1996

Polyphenol oxidase(PPO) isolated from the crude extract of ascidian, Halocynthia roretzi, showed higher affinity for catechol than tyrosine or DL-DOPA. Successful enzyme assay could be performed at $25^{\circ}C$, 10min. by mixing 0.2ml of crude enzyme extract with 2.8ml of 0.13M catechol in 0.1M sodium phosphate buffer(pH 6.4). The specific activity of PPO which had been purified with a combination of ammonium sulfate treatment, ion exchange chromatography on DEAE-cellulose, and gel filtration on Sepharose 6B was 13-fold disc gel electrophoresis. The activity of PPO was stable from pH 5.0 to 8.0 and showed the peak activity at pH 6.4 .The optimum reaction temperature for PPO oxidation on catechol was 35$^{\circ}C$ and those enzyme were heat stable up to 4$0^{\circ}C$. Molecular weigth of the enzyme was estimated about 170kDa. One molecule was found to be composed of gour subunits. Two of them had molecular weigh of 55kDa and the others 30kDa. The {TEX}$K_{m}${/TEX} values, {TEX}$V_{max}${/TEX} and catalytic efficiency({TEX}$V_{max}${/TEX}/{TEX}$K_{m}${/TEX}) for catechol were 0.12mM, 2.5mM/liter/min. and {TEX}$0.18min^{-1}${/TEX} respectively. The substrate affinity and electrophorectic pattern suggested that the enzyme of ascidian was considered to be not tyosine but catechol oxidase.

• Ocean and Polar Research
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• v.40 no.4
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• pp.249-257
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• 2018

Matrix metalloproteinases (MMPs) are associated with the invasion and metastasis of malignant tumors composed of cancer cells in an increased state of expression. This study evaluates the inhibitory effect of Carex pumila on MMP-2 and MMP-9 activity in phorbol-12-myristate-13-acetate (PMA)-stimulated HT-1080 human fibrosarcoma cells using gelatin zymography, MMPs enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay. C. pumila was extracted twice with dichloromethane ($CH_2Cl_2$) and methanol (MeOH). Treatment with $CH_2Cl_2$ extract and MeOH extract in PMA-stimulated HT-1080 cells effectively reduced the production of MMP-2 and 9. Also, the combined crude extracts ($CH_2Cl_2$ and MeOH) significantly inhibited the enzymatic activities and the expression of MMP-2 and MMP-9 in mRNA and protein levels. The combined crude extracts were partitioned between $CH_2Cl_2$ and water. The organic layer was further fractionated with n-hexane, 85% aqueous methanol (85% aq.MeOH) and the aqueous layer was separated into n-butanol and water, successively. Of the fractions, 85% aq.MeOH fraction showed the highest inhibitory activity of MMP-2 and MMP-9 in gelatin zymography and MMP ELISA kit. Furthermore, 85% aq.MeOH fraction most significantly suppressed cell migration. In RT-PCR and Western blot assay, n-butanol and 85% aq.MeOH fractions exerted the greatest inhibition on mRNA and protein expression of MMP-2 and MMP-9, respectively. As a result, C. pumila can be used as a good anti-invasive agent source.

• Natural Product Sciences
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• v.27 no.4
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• pp.245-250
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• 2021

The methylglyoxal (MGO) trapping constituents from Malus baccata L. were investigated using incubation of MGO and crude extract under physiological conditions followed by HPLC analysis. The peak areas of MGO trapping compounds decreased, and their chemical structures were identified by HPLC-ESI/MS. Sieboldin was identified as a major active molecule representing MGO-trapping activity of the crude extract. After reaction of sieboldin and MGO, remaining MGO was calculated by microplate assay method using imine (Schiff base) formation of 2,4-dinitrophenylhydrazine (DNPH) and aldehyde group. After 4 h incubation, sieboldin trapped over 43.8% MGO at a concentration of 0.33 mM and showed MGO scavenging activity with an RC₅₀ value of 0.88 mM for the incubation of 30 min under physiological conditions. It was also confirmed that sieboldin inhibited the production of advanced glycation end products (AGE) produced by bovine serum albumins (BSA)/MGO. Additionally, MGO trapping mechanism of sieboldin was more specifically identified by ¹H-, ¹³C-, 2D NMR and, confirm to be attached to the position of C-3' (or 5').

• Korean Journal of Medicinal Crop Science
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• v.11 no.3
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• pp.224-231
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• 2003

In this study effect of different mulching materials on yield and crude saponin contents in Anemarrhena asphodeloides Bunge. The results of this study are summarized as follows: The rhizome yield of seed propagation type in transparent PE mulching cultivation was of which increased 16.7% more than those of non-mulching cultivation. The rhizome yield of young-plant propagation type in green PE mulching and transparent PE mulching cultivations were of which increased 29.4% and 26.5%, respectively more than those of non-mulching cultivation. The contents of methanal extract of seed propagation type in transparent PE mulching and black PE mulching cultivations were of increased each 31.0%, 15.4% more than those of non-mulching cultivation. Crude saponin contents of seed propagation type in transparent PE mulching cultivation was increased up to 30.3% compared to those of non-mulching cultivation. However, the crude saponin contents of seed propagation type in green PE mulching and black & white PE mulching cultivations were not affected by the non-mulching cultivation. Crude saponin contents of young-plant propagation type in transparent PE mulching cultivation was increased 24.0% and 15.4% more than those of green PE mulching and black PE mulching cultivations.

• Food Science of Animal Resources
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• v.30 no.6
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• pp.989-996
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• 2010

This study was aimed at investigating the differences in chemical composition of antler according to growing days (80 d and 90 d after casting) and sections (upper, middle, base) in elk. Antlers of six bulls (aged 3 years) cutted on day 80 and 90 after casting were used in this trial and compared chemical composition such as crude protein, crude fat, crude ash, calcium, magnesium, amino acids, lipids, and fatty acids. The contents of crude protein and ether extract were higher in antlers cutted on day 80 than in antlers cutted on day 90, but that of crude ash was higher in antlers cutted on day 90. Significantly difference was observed in crude protein content between growing days in middle part of antler (p<0.05). Calcium and magnesium contents were higher on 90 d than 80 d, but there was no significant difference between growing days of antler. Based on the growing period of antler, there were significant differences (p<0.05) in aspartic acid, threonine, valine, leucine, phenylalanine and lysine, and essential amino acids content was higher in antlers cutted on day 80. Total lipid, neutral lipid, and phospholipid contents were higher in antlers cutted on day 80 than in those cutted on day 90, sphingo-phospholipid content was higher than glycerophospholipid content and values was higher in antlers cutted on day 80 than in those cutted on day 90. For total fatty acids, unsaturated fatty acids composition, there was no significant difference between growing periods of antlers, but values was higher on day 80 than on day 90. Essential fatty acids contents were lower in antler cutted on day 90 than antlers cutted on day 80 and higher for upper part than lower part.