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Establishment of rhizosphere microbes for plant protection on soil-borne diseases -Benificial antagonist and its mode of action toward ginseng root rot pathogen- (근권미생물과 토양병방제 -유용길항균이 인삼근부병원에 미치는 영향-)

  • Kim, S.I.;Lee, M.W.
    • The Korean Journal of Mycology
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    • v.22 no.1
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    • pp.50-61
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    • 1994
  • From soil samples, 380 antagonistic microorgnisms were isolated. Among the isolates, 42 strains had mycelia growing inhibition ability against Fusariun solani, ginseng root rot causing pathogen. Isolates CHA 1 and S-PFHR 6 were proposed as antagonists for this study and they were identified as Promicromonospora sp. and Pseudomonas pseudoalcaligenes respectively. As an antagonism against hyphae of F. solani in dual culture test, CHA 1 and S-PFHR 6 inhibited linear growing, caused abnormal branching, and the membrane projection which formed by cell wall destruction. The secondary metabolites contained in the culture filtrates which prepared from PD broth and Nutrient broth inhibited the spore germination to 14.3%. The culture filtrate of S-PFHR 6 which prepared by a little amount of soil extract addition to nutrient rich medium had more strongly. inhibited the spore germination and spore germination decreased to less than 4.0% in it. The soil used in this study had fungistasis and the germination rate of macroconidia and chlamydospore of F.solani was 19.4% and 17.7% respectively. The steam sterilized soil lost fungistasis and germination rate of conidia increased to more than 97.9%. The soils amended with the propagule of CHA 1 and S-PFHR 6 increased fungistasis and the germination rate of macroconidia decreased to 14.7% and 11.7% respectively in each treatments. But the soil ammended with glucose and asparagine annulled fungistatic ability and the germination rate of macroconidia increased to more than 48.0%. As an antagonistic activity of the secondary metabolites of two antagonistic isolates in soil, the germination rate of macroconidia of F. solani was 9.3% in the soil amended with the culture filtrate of CHA 1 but the culture filtrate of S-PFHR 6 had no such activity. In the soil which treated with antagonist propagule or culture filtrate, the chlamydospore germination rate was lower than that in natural soil. The addition of glucose and asparagine to antagonist propagule treated soil did not enhanced the chlamydospore germination.

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Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli (대장균이 생산한 재조합 인체 감마인터페론의 발현과 정제)

  • Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
    • KSBB Journal
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    • v.21 no.3
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    • pp.204-211
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    • 2006
  • For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

Metalloproteinase Plays a Role in Mucin Secretion (Mucin 분비에 영향을 미치는 Metalloproteinase)

  • Oh, Yeon-Mok;Choi, Hee Jin;Shim, Tae Sun;Lee, Sang Do;Kim, Woo Sung;Kim, Dong-Soon
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.3
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    • pp.289-296
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    • 2004
  • Background : Mucus hypersecretion in the patients with airway diseases represents poor prognosis as well as discomfort. However, there is no known therapy for its effective control. One important component of mucus is mucin, a glycosylated protein, which endows mucus with viscosity. We studied whether a proteinase has a role in mucin secretion and if so, which. Methods : (1) Inhibition of mucin secretion Group-specific proteinase inhibitors were tested to evaluate whether a proteinase belonging to a group of proteinases plays a role in mucin secretion. Phenylmethylsulfonyl fluoride(PMSF, a serine proteinase inhibitor), E-64(a cysteine proteinase inhibitor), Pepstatin(an aspartic proteinase inhibitor) and 1, 10-Phenanthroline(a metalloproteinase inhibitor) were treated into the Calu-3 cell line for 24 hours. The enzyme linked immunoabsorbant assay(ELISA) for MUC5AC was performed to evaluate the amount of mucin secretion and to compare with a control. (2) Stimulation of mucin secretion Matrix metalloproteinase-9(MMP-9), MMP-12 and TACE(TNF-alpha converting enzyme), which are known to be related with airway diseases, were used to be treated into Calu-3 for 24 hours. ELISA for MUC5AC was performed to evaluate the amount of mucin secretion and to compare with the controls. Results : (1) PMSF($10^{-4}M$), E-64($10^{-4}M$), Pepstatin($10^{-6}M$) and 1, 10-Phenanthroline($10^{-4}M$) reduced the MUC5AC secretion by $1{\pm}4.9%$(mean${\pm}$standard deviation; P=1.0 compared with the control), $-6{\pm}3.9%$(P=0.34), $-13{\pm}9.7%$(P=0.34) and $41{\pm}8.2%$(P=0.03), respectively. (2) The amounts of MUC5AC secretion stimulated by MMP-9(250ng/ml), MMP-12(100ng/ml) and TACE(200ng/ml) were $103{\pm}6%$(P=0.39), $102{\pm}8%$(P=1.0) and $107{\pm}13%$(P=0.39), respectively, compared with the controls. Conclusion : Metalloproteinase(s) is (are) suggested to play a role in mucin secretion. It appears that metalloproteinases, other than MMP-9, MMP-12 or TACE, affect the mucin secretion in this in vitro model.

Factors Predicting the Development of Radiation Pneumonitis in the Patients Receiving Radiation Therapy for Lung Cancer (방사선 치료를 시행 받은 폐암 환자에서 방사선 폐렴의 발생에 관한 예측 인자)

  • An, Jin Yong;Lee, Yun Sun;Kwon, Sun Jung;Park, Hee Sun;Jung, Sung Soo;Kim, Jin whan;Kim, Ju Ock;Jo, Moon Jun;Kim, Sun Young
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.1
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    • pp.40-50
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    • 2004
  • Background : Radiation pneumonitis(RP) is the major serious complication of thoracic irradiation treatment. In this study, we attempted to retrospectively evaluate the long-term prognosis of patients who experienced acute RP and to identify factor that might allow prediction of RP. Methods : Of the 114 lung cancer patients who underwent thoracic radiotherapy between December 2000 and December 2002, We performed analysis using a database of 90 patients who were capable of being evaluated. Results : Of the 44 patients(48.9%) who experienced clinical RP in this study, the RP was mild in 33(36.6%) and severe in 11(12.3%). All of severe RP were treated with corticosteroids. The median starting corticosteroids dose was 34 mg(30~40) and median treatment duration was 68 days(8~97). The median survival time of the 11 patients who experienced severe RP was significantly poorer than the mild RP group. (p=0.046) The higher total radiation dose(${\geq}60Gy$) was significantly associated with developing in RP.(p=0.001) The incidence of RP did not correlate with any of the ECOG performance, pulmonary function test, age, cell type, history of smoking, radiotherapy combined with chemotherapy, once-daily radiotherapy dose fraction. Also, serum albumin level, uric acid level at onset of RP did not influence the risk of severe RP in our study. Conclusion : Only the higher total radiation dose(${\geq}60Gy$) was a significant risk factor predictive of RP. Also severe RP was an adverse prognostic factor.

Effect of Steroid Administration Ex Vivo on the IκB/NF-κB Pathway in Human Peripheral Blood Monocytes (스테로이드의 투여가 말초혈액 단핵구에서 IkB/NF-κB경로에 미치는 영향)

  • Yoon, Ho Il;Lee, Hee-Seok;Lee, Chang-Hoon;Lee, Choon-Taek;Kim, Young Whan;Han, Sung Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.5
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    • pp.542-550
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    • 2003
  • Background : Synthetic glucocorticoids are widely used in many chronic inflammatory diseases because of their excellent anti-inflammatory activity. Enhancing the transcription of $I{\kappa}B$ and preventing activated NF-${\kappa}B$ from binding to ${\kappa}B$ sites are thought to be the underlying mechanisms. But these data are largely derived from in vitro studies using cell lines. In this study, after administrating a steroid to volunteers, we evaluated the effect on the NF-${\kappa}B$ system. Methods : Prednisolone(0.5mg/kg/d) was orally administered to 5 healthy volunteers for 7 days. Before and after the administration, we sampled their peripheral blood monocytes, and performed western blot analysis both with stimulation, using IL-$1{\beta}$, LPS, TNF, and without stimulation(baseline). We also performed EMSA after stimulation with LPS. Results : After ingestion of the steroid, baseline expressions of $I{\kappa}B{\alpha}$ were increased in two of the subjects, while suppressed degradations of $I{\kappa}B{\alpha}$ to stimulations were observed in all five. In addition, the binding capacity of NF-${\kappa}B$ after the administration was decreased. Conclusion : Steroid plays such roles as enhancing the transcription of $I{\kappa}B{\alpha}$, suppressing the DNA binding capacity of NF-${\kappa}B$, and suppressing the degradation of $I{\kappa}B{\alpha}$.

Effect of L-Ascorbic Acid on Collagen Synthesis in 3T6 Fibroblasts and Primary Cultured Cells of Chondrocytes (3T6 세포주 및 연골 초대배양세포의 Collagen 합성에 미치는 비타민 C의 영향)

  • Kim, Mi-Hyang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.42-47
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    • 2006
  • L-Ascorbic acid (AsA) is an essential nutrient for prevention of scurvy in humans, primates and guinea pigs that lack $L-gulono-\gamma-lactone$ oxidase which is required for the final step of AsA biosynthesis. AsA participates in various hydroxylation reactions involved in the biosynthesis of collagen. The purpose of this study is to clarify the role of AsA on collagen synthesis in 3T6 fibroblasts and primary cultured cells of chondrocytes. Cells were cultured in medium supplemented with catalase and AsA at various concentration. Supplement of AsA induced collagen synthesis in 3T6 fibroblasts and primary cultured cells of chondrocytes. The most remarkable induction of collagen synthesis by AsA was found in primary cultured chondrocytes. The content of collagen representing the amounts of extracellular matrix significantly increased in the cells of which growth was stimulated by AsA, while it decreased with increasing passage numbers of subculture in cells. It showed that the content of collagen decreased in the medium which contained AsA at the concentration higher than 5.0 mM. However, the contents of collagen to DNA were not different among various AsA concentrations. Supplementing with AsA resulted in enhancement of collagen formation and extracellular matrix. Therefore, there might be a Positive correlation between the activity of catalase and the AsA concentration. Moreover, it can be assumed that AsA stimulates the collagen synthesis by optimizing the cell-culture environment.

Antibacterial Activities of Trace Elements in Combination with Food Additives (미량원소 강화 식품소재의 항균효과)

  • Kim, Bo-Mi;Mok, Jong-Soo;Oh, Eun-Gyoung;Son, Kwang-Tae;Shim, Kil-Bo;Cho, Young-Je
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.1
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    • pp.35-41
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    • 2006
  • Antibacterial activities of the trace elements in combination with the food additives were measured against 6 kinds of food-borne microorganisms such as Escherichia coli, Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus, Bacillus subtilis and Pseudomonas fluorescens. The difference of antibacterial activity was not shown among the kinds of food additives, such as dextrin, gelatin and collagen. Zn and Ge in combination with food additives had strong antibacterial effect. Especially, $1\%$ zinc acetate in combination with $1\%$ gelatin was more effective against P. fluorescens and S. aureus than against Bacillus sp., E. coli and V. parahaemolyticus. Minimum inhibitory concentration of zinc acetate in combination with $1\%$ gelatin appeared to be 0.5 mg/mL on S. aureus and P. fluorescens, and 1.0 mg/mL on E. coli, V. parahaemolyticus, B. cereus and B. subtilis. Minimum bactericidal concentration of zinc acetate in combination with $1\%$ gelatin appeared to be 0.5 mg/mL on P. fluorescens and 1.0 mg/mL on E. coli, V. parahaemolyticus, S. aureus, B. cereus and B. subtilis. The zinc acetate in combination with gelatin showed stronger inhibitory effect in acidic range below pH 6.0, and remained active even after heat treatment at $121^{\circ}C$ for 15 min. In comparison with control, the viable cell counts of fish pastes, which were coated with the solution containing both $1\%$ zinc acetate and $3\%$ gelatin, were decreased by more than 100-fold until the storage of 7 days at $10^{\circ}C$. The results indicate that the combined use of zinc acetate and some food additives could prolong the shelf life of fish pastes by 8 days or more at $10^{\circ}C$.

Protective Effect of a Herbal Preparation (HemoHIM) on the Self-Renewal Tissues and Immune System against γ-Irradiation (방사선에 대한 생약복합조성물(HemoHIM)의 재생조직 및 면역계 방호 · 회복촉진 효과)

  • Jo, Sung-Kee;Park, Hae-Ran;Jung, Uhee;Oh, Heon;Kim, Sung-Ho;Yee, Sung-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.805-813
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    • 2005
  • In our previous study, a novel herb mixture (HIM-I) of Angelim gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix was developed to protect the intestinal and immune systems and promote its recovery against radiation damage. In this study, a new herbal preparation (HemoHIM) with the high immune modulating activity was developed from HIM-I. HIM-I was fractionated into ethanol fraction (HIM-I-E) and polysaccharide fraction (HIM-I-P). And HemoHIM was prepared by adding HIM-I-P to HIM-I. The protective activities against $\gamma$ -irradiation were compared among HemoHIM, HIM-I and the fractions. HemoHIM and HIM-I significantly decreased the radiation-induced DNA damage in vitro, and scavenged hydroxyl radicals in a dose-dependent manner. HemoHIM showed similar activity to HIM-I. In vitro proliferation assay with mouse lymphocytes and bone marrow cells showed that HIM-I-P was remarkably higher than HIM-I and HIM-I-E in cell proliferating activity. HemoHIM showed higher activity than HIM-I and this might be associated with the higher polysaccharide content. The in vivo protective effects of HemoHIM and HIM-I were investigated in $\gamma$-irradiated mice. HemoHIM increased the surviving intestinal crypts to a similar extent compared with HIM-I. In contrast, HemoHIM appeared to be more effective than HIM-I in endogenous spleen colony formation assay. The recovery of white blood cells and lymphocytes in irradiated mice were significantly enhanced by the administration of HemoHIM. Also HemoHIM administration prolonged the survival of irradiated mice. These results showed that the novel herbal preparation, HemoHIM, effectively protected the self-renewal tissues and immune system, and promoted the survival of irradiated mice. Moreover, in comparison with HIM-I, HemoHIM maintained similar activity in the reduction of oxidative damage of self-renewal tissue but exhibited the higher activity in protection and proliferation of immune and hematopoietic cells. These results suggested that HemoHIM might be more effective than HIM-I in immune modulation as well as radioprotection.

The Physiological Efficacy of Aloe Gel (Aloe Gel의 생리 효과에 대한 고찰 - Gel의 다당류와 미량 성분을 중심으로 -)

  • 서화중
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.6
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    • pp.1026-1038
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    • 1995
  • Although aloe lost a lot of its previous popularity in modern clinical uses as medicine numerous scientific researches still have claimed the beneficial properties(curing and general tonic effect) of aloe gel. Whereas considerable contradictory reports have helped to confuse the aloe gel issue and continually aroused controversy about aloe gel efficacy. However health food, cosmetic and medicinal products made from aloe gel are widely available in the world market especially in U.S.A. so the growing of Aloe plant and the processing of A. vera gel have become big industries in some countries. In some previous papers the salicylic acid, one of the common trace gel components, was thought to have an analgetic and antinflammatory effect. Large amount of Mg ion in the gel was suggested to act as anesthetic, Mg-lactate as antihistamic, and Aloctin A(a glycoprotein) as wound healer by promoting the cell growth. The carboxypeptidase and bradykinase activity in the gel were proposed to have the pain relieving and antiinflammatory effect. But any of thes etheories concerining the physiological action of the trace gel components has not been demonstrated by modern pharmacology, and failed to be supported by clinical research. It was suggested by some research workers that trace amount of anthraquinone compounds in the gel play an important role to act as false substrate inhibitors for PG and TX production(antiprostanoid effect), by which, they believed, inflammation, burn and frostbite, and infected wound could be healed. This hypothesis has not been substantiated. Butthe suggested antimicrobial action, antidiabetic, and antidotic effect of aloe gel are likely to be attributed to the trace anthraquinone compounds. In a lot of recent experimental reports it has been claimed that aloe gel polysaccharides(acetylglucomannan, acetylmannan, and glycoprotein) have the antimicrobial, antinflammatory, antitumour, and infected wound healing effect by immunoenhancement. It is hoped that these effects will be soon documented in clinical studies, then the controversy on aloe gel beneficial effect will cease. In the 30 days subchronic toxicity test the lowest observed adverse effect level of acemannan(acetylmannan) on dog was 5.0 mg/kg, IP. But the aloe gel is generally agreed to be harmless and non toxic even for the internal use such as health food. In the case of idiosynrasy one must keep the delayed type hypersensitivity reaction of aloe gel in mind. In conclusion it seem to be impossible to simply refuse a lot of evidences made by research workers who have claimed aloe gel's beneficial effects and to deny the fact that there had been long therapeutic histories of Aloe plants.

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Solubility of Hydrogen Sulfide and Methane in Ionic Liquids: 1-Ethy-3-methylimidazolium Trifluoromethanesulfonate and 1-Butyl-1-methylpyrrolidinium Trifluoromethanesulfonate (1-Ethyl-3-methylimidazolium trifluoromethanesulfonate와 1-Butyl-1-methylpyrrolidinium trifluoromethanesulfonate 이온성 액체에 대한 황화수소와 메탄의 용해도)

  • Lee, Byung-Chul
    • Korean Chemical Engineering Research
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    • v.54 no.2
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    • pp.213-222
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    • 2016
  • Solubility data of hydrogen sulfide ($H_2S$) and methane ($CH_4$) in two kinds of ionic liquids with the same anion: 1-ethyl-3-methylimidazolium trifluoromethanesulfonate ([emim][TfO]) and 1-butyl-1-methylpyrrolidinium trifluoromethanesulfonate ([bmpyr][TfO]) are presented at pressures up to about 30 MPa and at temperatures between 303 K and 343 K. The gas solubilities in ionic liquids were determined by measuring the bubble point pressures of the gas + ionic liquid mixtures with various compositions at different temperatures using a high-pressure equilibrium apparatus equipped with a variable-volume view cell. The $H_2S$ solubilities in ionic liquid increased with the increase of pressure and decreased with the increase of temperature. On the other hand, the $CH_4$ solubilities in ionic liquid increased significantly with the increase of pressure, but there was little effect of temperature on the $CH_4$ solubility. For the ionic liquds [emim][TfO] and [bmpyr][TfO] with the same anion, the solubility of $H_2S$ as a molality basis was substantially similar, regardless of the temperature and pressure conditions as a molar concentration basis. Comparing the solubilities of $H_2S$ and $CH_4$ in the ionic liquid [emim][TfO], the solubilities of $H_2S$ were much greater than those of $CH_4$. For the same type of ionic liquid, the solubility data of $H_2S$ and $CH_4$ obtained in this study were compared to the solubility data of $CO_2$ from the literature. When compared at the same pressure and temperature conditions, the $CO_2$ solubility was in between the solubility of $H_2S$ and $CH_4$.