• Title/Summary/Keyword: inoculation studies

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Antimicrobial Activity of Hwangryunheadok-tang and Extracts of Scutellariae Radix, Phellodendri Cortex, Coptis Rhizome, and Gardenia Jasminoides against Salmonella Typhimurium (黃連解毒湯, 黃連, 黃芩, 黃柏, 梔子의 Salmonella Typhimurium에 대한 항균효과)

  • Kim, In-soo;Lee, Young-su
    • The Journal of Internal Korean Medicine
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    • v.36 no.2
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    • pp.165-179
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    • 2015
  • Objectives: This experiment investigated the antimicrobial activity of Hwangryunheadok-tang and extracts of Scutellariae radix , Phellodendri cortex , Coptis rhizome , and Gardenia jasminoides against Salmonella typhimurium . Methods: After spreading S. typhimurium on a bacterial culture medium plate, antimicrobial activity was tested by dripping diluted Hwangryunheadok-tang or extracts of Scutellariae radix , Phellodendri cortex , Coptis rhizome , or Gardenia jasminoides (80 μl, diluted 100, 50, 10, and 1%) onto the plate, followed by culture for 16 to 72 hours. The minimal inhibitory concentration (MIC) was tested by dripping the minimum dilution that showed antimicrobial activity (80, 60, 40, and 20 μl) and measuring the density. The antimicrobial activity of Hwangryunheadok-tang and four extracts showed continuous antibacterial activity against S. typhimurium throughout the experiment. Result: 1. S. typhimurium . (Standard Microorganism, ATCC) 1) The Hwangryunheadok-tang and extracts of Scutellariae radix , Phellodendri cortex , and Coptis rhizome showed antibacterial activity in the undiluted solutions and in 50% dilutions. Gardenia jasminoides extract showed potency only in the undiluted solution. The antimicrobial potency of the undiluted solution was increased when the volume of inoculation was increased, but no difference was noted when the culture time was extended. All the extracts showed antimicrobial potency against S. typhimurium . 2. S. typhimurium isolated from diarrhea patients 1) When compared to the standard microorganism experiment on S. typhimurium , the MICs of the five extracts were increased. However, whereas the antimicrobial potency of doxycycline was lost entirely against bacteria isolated from patients with diarrhea, the antimicrobial potency of all the extracts was diminished but did not disappear. 2) The antimicrobial activity of Hwangryunheadok-tang and four extracts was continuous even when the culture time was extended to 16, 24, and 72 hours. Conclusions: The Hwangryunheadok-tang and four kinds of extracts have antimicrobial activity against S. typhimurium . Comparison of a standard microorganism with S. typhimurium isolated from diarrhea patients showed that the antimicrobial activity of all the extracts was better than that of antibiotics. Further studies should focus on the value and benefits of Hwangryunheadok-tang , and the four kinds of extracts as clinical treatments.

Studies on the development of mushroom media for bottle culture in new Pleurotus ostreatus 'Miso' (신품종 '미소' 느타리버섯 병재배 배지개발)

  • Lee, Byung-Joo;Kim, Yong-Gyun;Kim, Hong-Kyu;Yang, Euy-Seog;Lim, Yong-Pyo
    • Journal of Mushroom
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    • v.8 no.1
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    • pp.37-40
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    • 2010
  • This study was carried out to develop the best medium of new oyster mushroom for bottle culture. The new oyster mushroom cv. Miso is a Pleurotus ostreatus developed at the Chungnam Agricultural Research & Extension Services. For the bottle culture media, poplar sawdust+beet pulp+cottonseed meal (5:3:2), poplar sawdust+wheat husk meal (8:2), poplar sawdust+beet pulp+cottonseed meal (6:2:2), poplar sawdust+beet pulp+cottonseed meal (4:4:2), and poplar sawdust+beet pulp+wheat husk meal (7:1:2) were used in 850cc PP bottle. The pH was 5~6 and the C/N ratio 19.7~28.3 in bottle culture media. The time of pinhead formation was 5 to 6 days. For the fruiting body formation after inoculation took 29~31 days. The yield of fruiting body of poplar sawdust+beet pulp+wheat husk meal (7:1:2) medium was the highest at 110.4g/bottle compared to other media. Therefore, such cultivation medium would be appropriate for the commercial production of bottle culture in the new oyster mushroom 'Miso'.

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Studies on the Change of Enzyme Activities during the Fermentation of Tobacco Leaves (엽연초(葉煙草) 발효중(醱酵中) 효소활성도(酵素活性度)의 변화(變化)에 관(關)한 연구(硏究))

  • Kim, H.S.;Chun, J.K.;Lee, S.R.;Bae, H.W.;Sung, H.S.
    • Applied Biological Chemistry
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    • v.8
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    • pp.75-80
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    • 1967
  • In order to investigate the change of enzyme activities during the fermentation of tobacco leaves, whole and cut leaves were treated for twenty days at $30^{\circ}C$ and $39^{\circ}C$ by the normal procedure and inoculation of yeast, respectively, and the activity of several enzymes was measured at proper intervals. 1) Alpha-amylase activity was steadily decreased with the progress of fermentation. 2) Invertase and catalase activities were steadily decreased until the mid of the period, and then increased again toward the end of the fermentation. The latter enzyme activity showed more marked change than the former. 3) Most of the polyphenol oxidase activity were decreased abruptly in the early stage of the fermentation, and thereafter leveled down slowly. 4) Peroxidase activity was detected throughout the fermentation but no remarkable change was observed. 5) Protease activtty was not detected through the fermentation period.

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Studies on the Cultural Characteristics of Poria cocos (복령(Poria cocos)의 배양학적(培養學的) 특성(特性)에 관한 연구(硏究))

  • Hong, In-Pyo;Lee, Min-Wong
    • The Korean Journal of Mycology
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    • v.18 no.1
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    • pp.42-49
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    • 1990
  • The cultural characteristics and some factors such as nutrient sources and supplements effecting on mycelial growth and density were investigated to study the possibility of an artificial cultivation of P. cocos. The optimum pH for P. cocos was 4.0-4.5. The optimal growth temperature ranged from $25^{\circ}C$ to $29^{\circ}C$. Myceial growth of P. cocos was better in SPD than PD media. Adding the nurient sources such as dextrose, yeast and potato infusion to pine extract media practically stimulated the mycelial growth and density of P. cocos comparing to pine extract media alone. When P. cocos was cultured on sawdust media added 3 different supplements composed of corn meal, rice bran and wheat bran, corn meal was the best and its percentage was 30 (w/w) for mycelial growth. On culturing in sawdust media added by varying the mixture ratio of them, the media mixed corn meal and wheat bran (3:1, w/w) supported more vigours for mycelial growth. In inoculation test to pine stem, the fungal growth was good in under or inside pine bark and xylem, but the sclerotium was not observed in the stem. Mycelial growth was also observed in central part of pine stem by cross section.

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Microbiological Studies and Biochemical Changes in Fermenting Soybean Curd Residue during Fermentation (비지의 발효과정중 발효미생물 및 성분변화)

  • Lee, Moon-Sook;Kim, Kil-Hwan;Lee, Gui-Ju
    • Korean Journal of Food Science and Technology
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    • v.19 no.6
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    • pp.520-527
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    • 1987
  • This study was attempted to identify microorganisms in fermenting soybean curd residues (SCR). The changes in contents of free amino acids, nucleosides, reducing sugars, and oligosaccharides were also studied. The fermentation of SCR which was by change inoculation was carried out at $55^{\circ}C$ for 48 hrs. pH increased gradually during fermentation and isolated microorganisms were identified as Bacillus subtilis and Bacillus licheniformis. Moisture content decreased from 80.8% to 58.4% at 48 hrs of fermentation and other proximate composition did not show any changes during fermentation. The content of total free amino acids increased rapidly and the number and quantities of each amino acid analyzed also increased during the course of fermentation. Glutamic acid, phenylalaine, lysine and aspartic acid were rapidly liberated during fermentation. As to the changes of nucleotides, 5'-AMP little changed during the first 36 hrs but subsequently decreased to approximately 1/6 after 48 hrs of fermentation. On the other hand, 5'-IMP plus 5'-GMP did not show almost any change during the first 36 hrs but increased about 3.5 times at 48 hrs of fermentation. However, 5'-XMP was not detected. The reducing sugar level showed rapid and steady increase throughout the fermentation and that of stachyose plus raffinose decreased slightly. From these results, a possible way of utilization of fermented SCR was proposed as a substitute for soybean in meju preparation.

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STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.9-22
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    • 1974
  • Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

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Studies on the Seed Transmission of Colletotrichum spp. in Red Pepper (Capsicum annuum) (고추 탄저병균(炭疽病菌)의 종자전염(種子傳染)에 관(關)한 연구(硏究))

  • Yu, Seung Hun;Park, Jong Seong;Lee, Hyang Burm;Kim, Hong Gi
    • Korean Journal of Agricultural Science
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    • v.14 no.1
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    • pp.16-25
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    • 1987
  • Colletotrichum acutatum, C. coccodes, C. dematium and C. gloeosporioides were detected in seed samples of red pepper (Capsicum annuum). C. dematium and C. gloeosporioides were the predominant species, maximum seed infection of the species in some samples were 84% and 28%, respectively. C. acutatum and C. coccodes were recorded only in low percentages of 1-2. The blotter method proved more suitable for detecting Colletotricum spp. than the deep freezing blotter or agar plate methods. Plating of seed components showed that C. dematium and C. gloeosporioides were recovered more frequently from seed coat, and decreasing amounts of infection were observed in the endosperm and cotyledon. Seed-borne C. dematium and C. gloeosporioides caused seed rot, damping-off, seedling blight and brown discoloration of cotyledon and hypocotyl when infected seeds were sown in agar of test tube or in soil. Inoculation experiments showed that C. acutatum was pathogenic to red fruit of red pepper and C. coccodes was highly pathogenic to red fruit and weakly pathogenic to leaf of the plant. C. dematium was highly pathogenic to leaf and green fruit and C. gloeosporioides was pathogenic to not only leaf but also green and red fruits. Host range of the four seed-borne species of Colletotrichum was also investigated.

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SYNERGISTIC APOPTOTIC EFFECT OF TAXOL ON ORAL SQUAMOUS CELL CARCINOMA BY CYCLOSPORIN A (구강 편평세포암종에서 Taxol과 Cyclosporin A의 세포사멸 상승 작용 효과)

  • Suh, Min-Jung;Han, Se-Jin;Lee, Jae-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.5
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    • pp.394-404
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    • 2007
  • Oral squamous cell carcinoma is the most prevalent oral cancer, which is characterized by its high metastasis and recurrent rates and poor prognosis. Taxol is an anticancer agent which is microbial products extracted from jew tree. It combines with the tubulin and induces apoptosis by inhibiting mitosis of cell with microtubule stabilization. Recently, it was reported to be effective in various solid tumors, but only very slight effect has been seen in oral squamous cell carcinomas due to its cell-specific potencies. Cyclosporin A is used as immune suppressant and is being applied in anticancer therapy as its mechanism of induction of change of apoptotic process in various cells have been known. In this study, oral squamous cell carcinoma HN22 cell line was used for in vitro experiment and as for the experimental group taxol and cyclosporin A were applied alone and to observe the synergistic effect of apoptosis, Taxol and cyclosporin A were coadministered with different concentration of taxol for comparison. The results were obtained as follow: 1. There was no difference in Bcl-2, Bax, caspase 3, 8, 9 mRNA expression when cyclosprin A or taxol was applied alone to HN 22 cell line. 2. Caspase 3, 9 mRNA expression was prominently increased when cyclosprin A and taxol were applied together to cancer cell. 3. No significant difference was observed when cyclosporin A and taxol($1{\mu}g/ml$ and $3{\mu}g/ml$) were applied together to cancer cell line. 4. No significant difference was seen in Bcl-2, Bax, and caspase 8 mRNA expression in all the groups of in vitro experiments. 5. When cyclosporin A was applied alone in vivo study on the nude mice, histopathologi cal findings was similar to those of the control group. Oral squamous cell carcinoma induced by inoculation of HN 22 cell line was not reduced after treatment of cyclosporin A. 6. When taxol was applied alone, the islands of squamous cell carcinoma still remained, which meant insignificant healing effect. There was a lesser volume increase compared with the cyclosporin A alone. 7. When taxol and cyclosporin A were applied together, the connective tissue and calcification were seen in the histopathologic findings. Oral squamous cell carcinoma was decreased and cancer cell was disappeared. In observing the tumor mass change with time, there was a gradual decreased size and healing features. As the results of the in vitro experiment, it could conclud that only when the two agents are applied together, mitochondria-mediated apoptosis occurred by considerable increase of caspase 3, 9 mRNA expression, irrespectable of the concentration of taxol. In vivo experiment, there was a discrete synergistic effect when the two agents were applied together. But single use of cyclosporin A was not effective in this study. Based on the results of this experiment, if further clinical studies are done, taxol and cyclosporin A could be effectively used in treatment of oral squamous cell carcinomas.

Studies on the Artificial Cultivation of Pleurotus cystidiosus O.K.Miller, the Abalone Mushroom (전복느타리버섯(Pleurotus cystidiosus O.K.Miller)의 인공재배에 관한 연구)

  • Jang, Kab-Yeul;Jhune, Chang-Sung;Shin, Chul-Woo;Park, Jung-Sik;Oh, Se-Jong;Choi, Sun-Gyu;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.31 no.3
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    • pp.200-205
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    • 2003
  • The study was carried out on the artificial cultivation of the abalone mushroom, Pleurotus cystidiosus O.K.Miller. The pine sawdust substrates with 20% rise bran were good for mycelial growth and high quantity of P. cystidiosus in the bottle cultivation. Moreover, the proper volume for bottle cultivation was 850 ml and the removal of spawn and surface layer of the medium before pin-heading was more efficient. The yields of P. cystidiosus were higher in sawdust substrates added calcium carbonate than those not added calcium carbonate. The volume of 3 kg polypropylene bag is good for yield and biological efficiency in bag cultivation of P. cystidiosus. Cotton wastes were proper substrates for bag cultivation. In the effect of different cultivation temperature, $28{\pm}2^{\circ}C$ cultivation temperature was good for for primordial formation after inoculation.

A Study on the Antitumor Activity of Panax ginseng (고려인삼의 항암효과에 관한 연구)

  • Hwang, Woo-lk
    • Journal of Ginseng Research
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    • v.17 no.1
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    • pp.52-60
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    • 1993
  • Panax ginseng has been extensively used in the traditional oriental medicine as a restorative, tonic and Prophylactic agent. Recently, several reports regarding to anticancer effects of Panax ginseng has accumulated. These studies emphasized the fact that the anticancer activities might be due to a glycoside group called ginsenoside or pan.u saponin which has a water soluble characteristic. However, the authors and collaborates demonstrated that a highly lipid soluble component in extract of Panax ginseng roots contains a considerable cytotoxic activities against marine leukemic cells (L1210, P388) and human censer cells (HRT-18, HT-29, HCT48). This study was devised to observe the cytotoxic activities of Petroleum-ether extract of Panax giuseng roots (crude GBD and its Partially Purified fraction from silicic acid column chromatography (7 : 3 GX) against sarcoma-180 (5-180) and Walker carcinosar- coma 256 (Walker 256) in vivo, and murine leukemic Lymphocytes (L1210) and human rectal cancer cells (HRT-18) and human colon cancer cells (HT-29 and HCT48) in vitro. Each cell-line was cultured in medium containing serial concentration of the crude GX or 7 : 3 GX in vitro. A highly lipid soluble compound in the extract of Panax ginseng root was cytocidal to murine leukemic cells and human colon and rectal cancer cells in vitro. In the meantime, ginseng saponin derivatives did not have cytotoxic effects at its corresponding concentration. The growth rates of the cancer cells in medium containing ginseng extracts were inhibited gradually to a significant degree roughly in proportion to the increase of the extract concentration. The cytotoxic activity of 7 : 3 GX was about 3 times more potent than that of crude GX, one unit of cytotoxic activity against L1210 cells being equivalent to 2.54 Ug and 058 Ug for the crude GX and 7 : 3 GX, respectively. The Ri value of the active compound on silica- gel thin layer chromatography with petroleum-ether/ethyl ether/acetic acid mixture (90 : 10 : 1, v/v/v) as a developing so lvent was 053. While, the Panaxydol and Panaxynol as active compounds were purified from Petroleum-ether extract of Panax ginseng root by Drs. Ahn and Kim, and author found out that the one unit of cytotoxic activity of the Panaxydol and Panaxynol against L1210 cells being equivalent to 056 Ug and 0.3918 respectively. The survival times of mice inoculated with S-180 cells were extended about 1.5 to 2 times by the 7 : 3 GX treatment compared with their control group. The significantly decreased hemoglobin values of rats after inoculation with Walker 256 were recovered to normal range by oral administration of the crude Gt The synthetic levels of protein, DNA and RNA in human colon and rectal cancer cells were significantly diminished by treatment with the crude GX, which can explain a part of the origin of its anticancer activity.

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