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Optimum Conditions of Freezing Lyophilization and Bioluminescence Activity Recovery for Environmental Applications Using a Recombinant Strain (유전자 재조합 균주를 환경에 적용하기 위한 (동결) 건조 및 활성회복 조건 최적화)

  • Ko Kyung-Seok;Kim Myung-Hee;Kong In-Chul
    • Journal of Soil and Groundwater Environment
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    • v.11 no.5
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    • pp.43-50
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    • 2006
  • Bioreporter bacteria, such as recombinant bioluminescent bacteria, have been used for the detection of specific compounds in complex environmental media. In this study, optimum conditions for the preparation and application of deep-freezed and Iyophilized recombinant bioluminescent strain KG1206 were investigated for the future application on contaminated environmental sites. Genetically engineered microorganism, Pseudomonas putida mt-2 KG1206, contains TOL plasmid and the plasmid inserted $P_{m}$, promoter on the upper part of lux gone in vector pUCD615, and m-toluate and benzoate are considered direct inducers for bioluminescence. Optimum conditions determined for the preparation and application of the deep-freezed and lyophilized strain were followings: cryoprotective agent (24% sucrose), lyophilization time (12 hrs), strain concentration ($OD_{600}=0.6$), reconstitution for freezed strain (quick reconstitution at $35^{\circ}C$), reconstitution for lyophilized strain ($3{\sim}6$ hrs exposure on LB medium), carrying conditions (keep at $20^{\circ}C$ after reconstitution). These results demonstrate the feasibility of deep-freezed or lyophilized state of genetically engineered bioluminescent strain for environmental usage.

Effects of SIS Sponge and Bone Marrow-Derived Stem Cells on the Osteogenic Differentiation for Tissue Engineered Bone (SIS 스폰지와 골수유래줄기세포를 이용한 조직공학적 골분화 유도)

  • Park Ki Suk;Jin Chae Moon;Yun Sun Jung;Hong Keum Duck;Kim Soon Hee;Kim Moon Suk;Rhee John M.;Khang Gilson;Lee Hai Bang
    • Polymer(Korea)
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    • v.29 no.5
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    • pp.501-507
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    • 2005
  • Small intestinal submucosa (SIS) had been widely used as a biomaterial without immune rejection responses. SIS sponges prepared by crosslinking with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). SIS powders dissolved in $3\%(v/v)$ acetic acid aqueous solution for 48hrs and freeze-dried. EDC solution ($H_2O$ : ethanol = 5 : 95) as a crosslink agent was used in concentration of 100mM. In vitro, rat-BMSCs seeded in SIS sponges and induced the osteogenesis for 28 days. We have characterized the osteogenic potential of rat-BMSCs in SIS sponges by alkaline phosphatase activity(ALP), n assay, SEM and RT-PCR for osteogenic phenotype. In SEM, all morphology of SIS sponges was regular and showed interconnected pore structure. By RT-PCR analysis, we observed type I collagen expression. These results demonstrate osteogenic differentiation of rat-BMSCs. In conclusion, we confirmed that the morphology of surface, cross-section, and side of SIS sponges were highly porous with good interconnections between each pores, which can support the surface of cell growth, proliferation, and differentiation. This result indicates that SIS sponge is useful for osteogenesis of BMSCs.

α-Glucosidase Inhibition Activity of the Extracts of Katsura Tree (Cercidiphyllum japonicum Sieb. Et Zucc) Leaves

  • Lee, Tae-Seong;Ryu, Wang-Gyun;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.43 no.2
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    • pp.238-247
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    • 2015
  • Katsura tree (Cercidiphyllum japonicum Sieb. Et Zucc) leaves were collected, air-dried and extracted with 70% aqueous acetone, then concentrated and sequentially fractionated using n-hexane, $CH_2Cl_2$, EtOAc, and $H_2O$ to be freeze dried for antioxidant and ${\alpha}$-glucosidase inhibition activity tests. The antioxidant activity of the extracts was evaluated using DPPH (1,1-diphenyl 2-picrylhydrazyl) free radical scavenging assay. The test concentrations were adjusted to 500, 250, 125, 62.5, 31.25, 15.62, 7.81, 3.9, 1.95 and 0.97 ppm. The $H_2O$ and EtOAc fractions showed higher activities compared with the control, ${\alpha}$-tocopherol, at all concentrations. The crude fraction also gave better activity at the concentrations lower than 62.5 ppm. However, the nonpolar n-hexane and $CH_2Cl_2$ fractions gave prominently lower activities compared with the control at all concentrations. The $IC_{50}$ values of the crude, EtOAc, and $H_2O$ fractions exhibited 11.78, 4.29 and $9.80{\mu}g/m{\ell}$, respectively, compared with $12.08{\mu}g/m{\ell}$ of the control. But the n-hexane and $CH_2Cl_2$ fractions indicated 300 and $91.85{\mu}g/m{\ell}$ of $IC_{50}$, respectively. ${\alpha}$-Glucosidase inhibition activity was evaluated at the concentrations of 50, 25, 12.5, 6.3, 3.1, 1.6 and 0.8 ppm. The inhibition activities were increased according to as the increase of sample concentrations. However, the nonpolar n-hexane and $CH_2Cl_2$ fractions indicated very low inhibition activities compared with acarbose, a positive control. The EtOAc fraction showed very good capability as almost 100% compared with the control at the higher concentrations than 12.5 ppm and the crude fraction also indicated good potential as 95% and 100% at 25 and 50 ppm, respectively. The $H_2O$ fraction gave good inhibition value as 90% at 50 ppm although the value was lower than the control. These results showed that the polar fractions had better ${\alpha}$-glucosidase inhibition activities. The $IC_{50}$ values of the nonpolar fractions, n-hexane and $CH_2Cl_2$, showed very lower values as 468 and $103.6{\mu}g/m{\ell}$, respectively, than the control. ${\alpha}$-Glucosidase Inhibition Activity of the Extracts of Katsura Tree (Cercidiphyllum japonicum Sieb. Et Zucc) Leaves However, the polar fractions, crude, EtOAc and $H_2O$, showed 7.1, 3.7 and $13{\mu}g/m{\ell}$, respectively, indicating that these fractions can be used as natural bioresources for treating diabetes mellitus. Also ${\alpha}$-glucosidase inhibition activity had a positive correlation with antioxidant activity of the extracts.

Quantitative analysis of rutin with mulberry leaves (II) (뽕나무 유전자원의 rutin 함량 변이(II))

  • Kim, Hyun-Bok;Kim, Sun-Lim;Lee, Sun-Ho;Sung, Gyoo Byung;Seok, Young-Seek;Kim, Yong Soon;Ju, Wan-Taek;Kim, Sung-Kuk;Chung, Da Eun;Jo, You-Young;Kweon, HaeYong;Lee, Kwang-Gill
    • Journal of Sericultural and Entomological Science
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    • v.52 no.2
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    • pp.129-133
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    • 2014
  • We analyzed rutin content using mulberry genetic resources. They were grown under the same environment and conditions. Mulberry leaves were collected and then freeze-dried and powdered for rutin test. As a result, mean content of mulberry strains was $0.38{\pm}0.17%$, and the coefficient of variation (CV) was 45.7%. The variation was greatly severe. Among the tested strains, 'Jangloe' was showed the highest content of 0.97%, whereas 'Sungsu 3' was showed the lowest content of 0.01% respectively. Finally we selected rutin high-containing 6 strains. They are as follows. 'Jangloe', 'Pumbo 6', 'Youjin', 'Kaeryangjeonjeon', 'Baekhak', and 'Pumbo 32' which are more than twice of the overall average content.

Screening for inhibitory effect on nine CYP isoforms by 20 herbal medications (고속 스크리닝 기법을 이용한 한약제제의 cytochrome P45O 저해능 탐색)

  • Kim, Hyun-Mi;Liu, Kwang-Hyeon
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.334-339
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    • 2007
  • We evaluated the potential of 20 herbal medications (HMs), commonly used in Korea, to inhibit the catalytic activities of several cytochrome P450 (CYP) isoforms. The abilities of 500 ${\mu}g/ml$ of aqueous extracts of 20 HMs to inhibit phenacetin O-deethylation (CYP1A2), coumarin 6-hydroxylation (CYP2A6), bupropion hydroxylation (CYP2B6), rosiglitazone hydroxylation (CYP2C8), tolbutamide 4-methylhydroxylation (CYP2C9), S-mephenytoin 4'-hydroxylation (CYP2C19), dextromethorphan O-demethylation (CYP2D6), chlorzoxazone 6-hydroxylation (CYP2E1), and midazolam 1'-hydroxylation (CYP3A) were tested using human liver microsomes. The HMs Woohwangcheongsimwon suspension and Hwanglyeonhaedok-Tang strongly inhibited CYP2B6 and CYP2D6 isoform activity, respectively. These results suggest that some of the HMs used in Korea have potential to inhibit CYP isoforms in vitro. Although the plasma concentrations of the active constituents of the HMs were not determined, some herbs could cause clinically significant interactions because the usual doses of those individual herbs are several grams of freeze-dried extracts.

Antibacterial Activities of hot-water and ethyl alcohol Extracts of Medicinal Herbs on Fish Pathogenic Bacteria (천연 생약재 열수 및 알코올 추출물의 어병 세균에 대한 항균력)

  • Choe, Hye-Seung;Kim, Lee-Cheong;Lee, Ju-Seok;Jo, Mi-Ra;Seo, Chang-Ho;Park, Su-Il
    • Journal of fish pathology
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    • v.17 no.1
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    • pp.39-55
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    • 2004
  • Hundreds of medicinal herbs have been using for the purpose of diseases treatment and immune enhancement for human being and other animals including fishes. Among them, 49 species of medicinal herbs were selected and tested for antibacterial activities against 19 strains of fish pathogenic bacteria in different 4 species. The 49 medicinal herbs were extracted by water and ethyl alcohol. The extracts were freeze dried and some paper discs from the extracts were prepared for the evaluation of antibacterial activity. The tested pathogenic bacteria were 5 strains of Edwardsiella tarda, 5 strains of Vibrio sp., 4 strains of Lactococcus garvieae, 1 strain of Lactococcus raffinose, 1 strain of Streptococcus parauberius, and 3 strains of Streptococcus iniae. The Galla rhois (Obaeja), Gaeonnamu and Hwangleyon showed antibacterial activities on both gram negative and gram positive fish pathogenic bacteria. The Youkgae, Sangbaekpi, Bogolji and Gamcho showed very effective antibacterial activities on gram positive pathogens while Jiyu, Aeyoeb and Yeonkyo showed very effective on gram negative pathogens.

The Effects of Freeze-Dried Potato Flour Addition on the Fermentation Characteristics of Yogurt (감마선 조사된 감자분말 첨가가 요구르트의 발효특성에 미치는 영향)

  • Ahn Byung-Young;Kim Dong-Han;Choi Dong-Seong
    • The Korean Journal of Food And Nutrition
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    • v.17 no.4
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    • pp.374-381
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    • 2004
  • This study was performed to characterize the mechanism of the acceleration of lactic acid fermentation in milk incubated with potato flour irradiated by ${\gamma}$-ray and two types of lactobacillus, Lactobacillus bulgaricus and Streptococcus thermophillus, and to examine its effect on the quality of yogurt by assessing the acidity of lactic acid, viscosity, free amino acid, and sensory. In lactic acid fermented in the presence of 0.5% (w/w) potato flour, its acidity was higher, its pH was lower, and its viscosity was increased than lactic acid fermented in the absence of potato flour. In the sample fermented with the mixture of S. thermophillus and L. bulgaricus, the acid production rate was highest. In the samples fermented with a single type of bacteria, the acid production rate of S. thermophillus was higher than that the L. bulgaricus. Fermented a single type of bacteria with potato flour, the lysine utilization rate of L. bulgaricus and S. thermophillus was high. L. bulgaricus produced a large quantity of glutamic acid whereas S. thermophillus consumed glutamic acid. In sensory evaluation of yogurt fermented with potato flour, the preference of texture was improved while its color, taste, flavor, and overall preference were decreased. The data suggest that L. bulgaricus stimulates the growth of S. thermophillus by providing free glutamic acid that is required by S. thermophillus and thus the addition of potato flour shortens the fermentation period of yogurt.

The Antioxidant Ability of Grape Seed Extracts (포도씨 추출물의 항산화성)

  • Jang, Jae-Kweon;Han, Ji-Young
    • Korean Journal of Food Science and Technology
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    • v.34 no.3
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    • pp.524-528
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    • 2002
  • The study was carried out to evaluate the antioxidant activity of grape seed extracts by measuring acid value, peroxide value (POV), thiobarbituric acid reactive substances (TBARS value) and electron donating ability by 1,1-dipheny1-2-pycryl hydrazil (DPPH) method. Total phenol content of freeze dried crude solvent extracts increased in the order of ethanol>acetone>hot water. Among extracts, the total phenol content of 70% ethanol extract of grape seed (GSE) was the highest, 51%. Besides the total phenol, the other major components of GSE were crude carbohydrate (29.63%), crude protein (3.38%) and crude lipid (2.84%). The acid values of crude rice bran oil containing GSE or several antioxidants at the concetration of 0.01% were 52.3 for the control, 0.5 for GSE, 2.3 for dibutyl hydroxytoluene (BHT), 45.0 for tertiarybutyl hydroxy quinone (TBHQ), 29.9 for tocopherol and 37.7 for ascorbic acid. The POVs for linoleic acid methyl ester in the presence of 0.01% antioxidants were 1220 meq/kg in control, 55 meq/kg in GSE, 104 meq/kg in BHT, 952 meq/kg in tocopherol and 71 meq/kg in GSE, 37% in BHT, 37% in tocopherol and 52% in ascorbic acid. The electron donating abilities by DPPH in the presence of 0.01% antioxidants were 95.3% in GSE, 75.0% in BHT, 96.3% in tocopherol and 98.2% in ascorbic acid. These results indicated that the antioxidants activities of GSE were significantly higher than those of several antioxidants compared.

Effects of Kimchi Supplementation and/or Exercise Training on Body Composition and Plasma Lipids in Obese Middle School Girls (운동 및 김치 보충제 섭취가 비만 여중생의 신체조성 및 혈중지질에 미치는 영향)

  • 백영호;곽정록;김세종;한성섭;송영옥
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.906-912
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    • 2001
  • The effects of aerobic exercises and/or supplementation of kimchi on changes of the body composition and plasma lipids of obese middle school girls were studied. Thirty eight girls, 28 obese girls and 10 normal weighed girls, were paricipated. Among obese girls, 8 were assigned to exercise group (FG) 12 were grouped as kimchi group (KG) , and 8 were asked to practice excercise and to take kimchi simultaneously(excercise kimchi group, EKG). Ten girls whose weight is normal asked to remain o their own diet during 6 weeks of experiment (control group, CG)/ EG practiced jogging and rope-jumping for 60 minutes four times a week and KG took 3 g of freeze-dried kimchi packed in a 500 mg capsule daily which is equivalent to 30 g of fresh kimchi, EKC, EG and KG showed beneficial effects on changes of the body composition and plasma lipids compared to those of CG, EG showed greater effect than KG in reducing body fat resulted decrease in BMI, fat mas,. abdominal fat, and triglyceride concentration and increase in HDL-cholesterol. KG seemed to have greater effect on lowering plasma cholesterol and LDL-cholesterol than EG/ But the greatest effects in terms of reduction in weight, BMI fat mass, abdominal fat, total cholesterol, LDL-cholesterol, triglyceride, and increase in HDL-cholesterol were observed from EKG. These results indicate that kimchi supplemenation while practicing excercise might improve the obese state by reducing body fat content as well as reducing plasma lipids.

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In vitro Cytotoxic Effect of Extracts from Styela plicata (오만둥이(Styela plicata) 추출물의 in vitro 세포독성 효과)

  • Lee, Bo-Bae;Cha, Mi-Ran;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.9
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    • pp.1099-1105
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    • 2007
  • The present study describes the preliminary evaluation of the anticancer activity of Styela plicata. Freeze-dried S. plicata was extracted with methanol, ethanol, acetone, and water, and then anticancer effect of the extracts was measured by the MTT reduction assay and phase-contrast microscopy on the HT-29 human colon carcinoma cells. Among the extracts, acetone extract showed the highest anticancer activity. The cell proliferation rates markedly decreased by 94.0% at the concentration of 500 ${\mu}g/mL$ of acetone extract compared with control cells. The acetone extract was further fractionated with hexane, diethyl ether, ethyl acetate, and water layer according to the degree of polarity. The HT-29 cells with hexane layer extract (250 ${\mu}g/mL$) decreased the cell viability to 5.1% of untreated control. The growth of SW620, HeLa, and MCF-7 cells was decreased to about 10%, by the treatment of hexane layer extract 250 ${\mu}g/mL$. Theses results suggest extracts from S. plicata as possible natural cancer therapeutic material.