In order to increase the content of germanium in ginseng adventitious roots, the effects of chelating agents on germanium content and root growth were investigated in the submerged cultures of ginseng adventitious roots. Chelating agents such as citric acid, oxalic acid, phosphoric acid, EDTA (Ethylenediamine tetraacetic acid) or EGTA (Ethylene glycol-bis $({\beta}-aminoethylether)-tetraacetic$ acid) were administrated in the submerged culture of ginseng root containing 50 ppm $GeO_2$. After 6 weeks of cultivation, fresh weight, germanium and saponin contents in the roots were analyzed. Among chelating agents, addition of 1.0mM phosphoric acid was found to be best for germanium accumulation. Under this condition, germanium content increased 1.4 times as compared to that of the control. The germanium content in the adventitious roots also increased with addition of EDTA or EGTA, while they inhibited the growth of ginseng adventitious root. Citric and oxalic acids were not effective for increasing germanium content in adventitious roots. As the results, it suggests that the phosphoric acid can be proved as the optimal agent for the enhancement of germanium accumulation in ginseng adventitious roots. These results can be served as a guideline for the mass production of ginseng adventitious roots containing germanium by large-scale production.
Fusarium proliferatum KGL0401 was previously isolated from Physalis alkekengi var. francheti plant roots and exhibited higher GA productivity than wild type Gibberella fujikuroi. The :tim of this work was to find out an optimal culture condition for GA production. Various carbon(fructose, glucose, lactose, maltose, sucrose) and nitrogen($KNO_3$, urea, glycine, $NaNO_3,\;NH_4Cl$) sources were used for this study. GAs activities were analysed by gas chromatography and mass spectrometry(GC-MS). The highest yield of $GA_3$ was found in the growth medium supplemented with sucrose as carbon source and $NH_4Cl$ as nitrogen source. The optimum carbon-nitrogen concentration for $GA_3$ production was found to be 0.5 M:0.17 M. Supernatant was prepared from the culture fluid of F. proliferatum KGL0401 cultured for 7 days at 3 0'E and the 10 ul supernatant was treated with 2 leaf-rice seedling.
The strain No. 33, which produces cellulose-degrading enzyme, was isolated from soil. Yellow halo was identified when the culture supernatant of the strain was loaded onto agar plate containing 2.0% CMC using paper disc method. From scanning electron microscopic observation, the morphology of the stain was rod-shaped. For identification, several biochemical characteristics were tested, and this strain was identified to Bacillus sp. So, we named this strain as Bacillus sp. No. 33. The maximal growth was observed when the stain was cultured in the medium containing 1.0% glucose, 3.0% yeast extract, 0.5% $KH_2PO_4$, 0.02% $MgSO_4{\cdot}7H_2O$, pH 7.0 at $30^{\circ}C$ for 39 hours with shaking. The maximal enzyme production was accomplished using the medium containing 4.0% CMC, 2.0% yeast extract, 0.5% $KH_2PO_4$, 0.04% $MgSO_4{\cdot}7H_2O$, pH 7.0 at $30^{\circ}C$ for 42 hours with shaking.
This study was performed to evaluate the effects of extracts of Drynariae Rhizoma on the characteristics of rat calvaria cells(RCV) and bone marrow cells(RBM) which have the important role on the bone formation in vitro. Drynariae Rhizoma has been known as the useful herbal medicament for treatment of the wound healing including regeneration of bone fracture, and also has been used to treat the periodontal lesions, tooth mobility, gingival bleeding and pus discharge via sulcus in Oriental Medicine. In control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 10% fetal bovine serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, extracts of Drynariae Rhizoma(0.1, 1, 5, 10, $50{\mu}g/ml$) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 3, 7, 14, 21, 30th day, the amount of total protein synthesis and alkaline phosphatase activity of RCV at 2,4th day and those of RBM at 3, 6th day. And also, the calcified nodule of RCV was examed at 3, 5th day in three goup, control, experimental, culture with the PDGF group. The results were as follow ; 1. Both RCV and RBM cells in Drynariae Rhizoma-treated experimental group proliferated more rapidly than nontreated control group. The experimental group below $5{\mu}g/ml$ Drynariae Rhizoma-treated showed more prominent cell proliferation from the 7th day to the 21st day than the control group and above $10\;{\mu}g/ml$ treated group in RCV. 2. Amount of total protein synthesis was more increased in Drynariae Rhizomatreated group than in control group. In $5{\mu}g/ml$ Drynariae Rhizoma-treated group showed most prominent protein synthesis of the any other exrperimental group and control group. 3. Alkaline phosphatase activity also more increased in Drynariae Rhizomatreated group than control group. 4. Mineralized nodules in Drynariae Rhizoma-treated group were more than not in control group but also in PDGF-treated group. From the above results, Drynariae Rhizoma appeared to enhanced the proliferation, protein synthesis, alkaline phosphatase activity and cellular ability of mineralized nodule formation than PDGF. So that, we conclude that Drynariae Rhizoma enhances the activities of bone cells which have the important role on the periodontal regeneration and optimal application of Drynariae Rhizoma was thought to be useful as the means in bone regeneration.
This study was performed to evaluate the effects of platelet-derived growth factor (PDGF) and epidermal growth factor (EGF) on the characteristics of beagle dog's periodontal ligament (BPD) cells and bone marrow (BBM) cells which have the important role on the early stage of periodontal tissue regeneration in vitro. In control group, the cells ($1.5{\times}10^5$cells/ml) were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum, $50{\mu]g/ml$ ascorbic acid, and 10mM/ml ${\beta}-glycerophosphate$. In experimental groups, growth factors, PDGF or EGF(10ng/ml), were added into the above culture condition. And then each group was characterized by examining the cell proliferation rate, amount of total protein synthesis, alkaline phosphatase activity at 1, 5, 9, 13, 17th day after seeding of cells into the culture wells. The results were as follows: 1. Both BPD and BBM cells in PDGF-treated group proliferated more rapidly than non-treated cells. This finding also was observed in EGF-treated group but it was not as prominent as that of PDGF-treated group. The proliferation rates of both cells showed the time-dependent pattern during experimental periods in all three groups. 2. Amount of total protein synthesis was more increased in PDGF-treated group than in control group. But no significant difference between EGF-treated group and control group was observed throughout experimental periods even though the tendency of amount of protein synthesis was time-dependent pattern. 3. Alkaline phosphatase activity also more increased in PDGF-treated group than control group. But slight decrease tendency was seen in both cells of EGF-treated group. From the above results, PDGF appeared to enhance the proliferation and cellular activities including amount of total protein synthesis and alkaline phosphatase activity of BPD and BBM cell, but EGF did not show notable effects. The optimal application of these growth factors was thought to be useful as the adjunctive means in periodontal regeneration procedures.
This study was carried out to determine the optimal medium composition and growth regulators for the micropropagation of Platycodon grandiflorum (Jacq.) A. DC. Nodes containing yellow green petals were used as plant materials to execute the study. The best performance of adventitious root development was found in 1/4 strength of MS basal salt and the growth was satisfactory in the concentration of 1/2 MS medium. The best condition for adventitious root development and growth was observed in the higher concentration (5%) of sucrose and activated charcoal free 1/4MS medium respectively. Adventitious roots were developed at the controlled culture medium at pH 4.8 with a tendency of suppression with higher levels of pH. However, it was prevailed that the development and growth depended on the concentration of agar. The lower concentration of agar (0.4%) was performed better than that of higher concentration (1.2%), whereas the agar concentration (0.4%) showed the best performance for the development and growth of adventitious roots. For the development of shoots containing node, BA combined with IAA was more effective than kinetin with IAA or NAA. The highest shoot development (3.9 shoots per explant) was performed on MS medium supplemented with 0.1 mg/L BA and 0.5 mg/L IAA.
Lim Kyoung-Suk;Son Shung-Hui;Kang Ho Young;Jun Hong-Ki
Journal of Life Science
/
v.15
no.4
s.71
/
pp.657-663
/
2005
Collagenases are generally defined as enzymes that are capable of degrading the polypeptide backbone of native collagen under conditions that do not denature the protein. An extracellular collagenase-producing bacterial strain was isolated from kimchi and identified to be Bacillus subtilis JS-17 through morphological, cultural, biochemical characteristics and 16S rDNA sequence analysis. Optimum culture condition of Bacillus subtilis JS-17 for the production of collagenase was $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4\cdot7H_2O,\;0.01\%\; MnSO_4\cdot4H_2O,\;,0.1\%$ citrate and $0.1\%\;CaCl_2$. The production of collagenase was optimal at $30^{\circ}C$ for 72 hr. A collagenase was isolated from the culture filtrate of Bacillus subtilis JS-17. The enzyme was purified using Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography and DEAE-Sephadex A-50 column chromatography The purified collagenase has an specific activity 192.1 units/mg. The molecular weight of the purified enzyme was estimated to be 28 kDa by SDS-PACE. The purified collagenase has $100\%$ activity up to $55^{\circ}C$.
Journal of the Korea Academia-Industrial cooperation Society
/
v.17
no.9
/
pp.322-328
/
2016
This study investigated the effects of temperature, salinity, and algal diet to find the optimal conditions for 5 days for the mass culture of the tropical tintinnid, Metacylis tropica. This tintinnid had a small, hyaline, and ovoid lorica. The oral diameter, length, and maximum width of the lorica were $36.7{\mu}m$, $49.5{\mu}m$, and $44.5{\mu}m$, respectively. In the temperature experiments, the highest maximum density and population growth rate were observed at $30^{\circ}C$ with 340.7 cells/mL and 1.1/day, respectively. Lower salinities adversely affected the population growth of M. tropica. The maximum density was observed at 33 ppt (840 cells/mL). In the diet experiments, M. tropica fed Isochrysis galbana showed the highest density (413 cells/mL) and population growth rate (1.2/day). As a result, M. tropica is appropriate as a potential prey organism for early fish larvae with smaller mouths because the tintinnid has a relatively small size compared to the rotifer. In addition, the conditions of $30^{\circ}C$, 33 ppt and supplying I. galbana would be effective in the cultivation of M. tropica.
Kim, Ji Min;Shin, Sung-Chan;Kwon, Hyun-Keun;Cheon, Yong-Il;Ro, Jung Hoon;Lee, Byung-Joo
Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
/
v.32
no.1
/
pp.15-23
/
2021
Background and Objectives During speech, the vocal folds oscillate at frequencies ranging from 100-200 Hz with amplitudes of a few millimeters. Mechanical stimulation is an essential factor which affects metabolism of human vocal folds. The effect of mechanical vibration on the cellular response in the human vocal fold fibroblasts cells (hVFFs) was evaluated. Materials and Method We created a culture systemic device capable of generating vibratory stimulations at human phonation frequencies. To establish optimal cell culture condition, cellular proliferation and viability assay was examined. Quantitative real time polymerase chain reaction was used to assess extracellular matrix (ECM) related and growth factors expression on response to changes in vibratory frequency and amplitude. Western blot was used to investigate ECM and inflammation-related transcription factor activation and its related cellular signaling transduction pathway. Results The cell viability was stable with vibratory stimulation within 24 h. A statistically significant increase of ECM genes (collagen type I alpha 1 and collagen type I alpha 2) and growth factor [transforming growth factor β1 (TGF-β1) and fibroblast growth factor 1 (FGF-1)] observe under the experimental conditions. Vibratory stimulation induced transcriptional activation of NF-κB by phosphorylation of p65 subunit through cellular Mitogen-activated protein kinases activation by extracellular signal regulated kinase and p38 mitogen-activated protein kinases (MAPKs) phosphorylation on hVFFs. Conclusion This study confirmed enhancing synthesis of collagen, TGF-β1 and FGF was testified by vibratory stimulation on hVFFs. This mechanism is thought to be due to the activation of NF-κB and MAPKs. Taken together, these results demonstrate that vibratory bioreactor may be a suitable alternative to hVFFs for studying vocal folds cellular response to vibratory vocalization.
This study was carried out to investigate the artificial cultivation of Pleurotus eryngii on the optimal medium vessel, periods of cultivation and the optimal method of pinheading for both yield and quality of fruiting body were also performed. The optimal composition of sawdust medium in polypropylene(PP) bottle was combination of sawdust(70%) and corncob(30%) but increased amount of corncob delayed the period of mycelial growth. The mycelial growth and the yield of fruiting body in the medium with beat pulp were worse than that without beat pulp. The optimal composition of nutrients for both yield and quality of fruit body tuned out to be a combination of rice bran(12%), wheat bran(12%) and cottonseed cake(6%). Additions of zeolite, shell lime and bean curd dregs were not effective in mycelial growth and yield of fruit body. When testing size of PP bottle for cultivation, the larger of bottle mouth is, the more pinheading number found, but the number of available fruit body is not significantly different. The culture in $1100\;ml-{\phi}75\;mm$ bottle is the best in the yield and quality of fruit body than those in $555\;ml-{\phi}50\;mm,\;850\;ml-{\phi}58\;mm,\;850\;ml-{\phi}65\;mm\;and\;1100\;ml-{\phi}65\;mm$ bottle. Using the PP bag for cultivation, a square shaped bag was better than a round shaped and black square shaped in mycelial growth and yield of fruit body. The most suitable period of incubation was 35 days after inoculation at $22{\pm}2^{\circ}C$. When the incubation periods was decreased less than 35 days, the pileus formation and yields were very bad but a pinheading condition looked similar, For an optimal pinheading, turning upside down was better than standing and covering.
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