• Title/Summary/Keyword: suspension-cultured-cells

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The Effects of Physicochemical Factors and Cell Density on Nitrite Transformation in a Lipid-Rich Chlorella

  • Liang, Fang;Du, Kui;Wen, Xiaobin;Luo, Liming;Geng, Yahong;Li, Yeguang
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2116-2124
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    • 2015
  • To understand the effects of physicochemical factors on nitrite transformation by microalgae, a lipid-rich Chlorella with high nitrite tolerance was cultured with 8 mmol/l sodium nitrite as sole nitrogen source under different conditions. The results showed that nitrite transformation was mainly dependent on the metabolic activities of algal cells rather than oxidation of nitrite by dissolved oxygen. Light intensity, temperature, pH, NaHCO3 concentrations, and initial cell densities had significant effects on the rate of nitrite transformation. Single-factor experiments revealed that the optimum conditions for nitrite transformation were light intensity: 300 μmol/m2/s; temperature: 30℃ pH: 7-8; NaHCO3 concentration: 2.0 g/l; and initial cell density: 0.15 g/l; and the highest nitrite transformation rate of 1.36 mmol/l/d was achieved. There was a positive correlation between nitrite transformation rate and the growth of Chlorella. The relationship between nitrite transformation rate (mg/l/d) and biomass productivity (g/l/d) could be described by the regression equation y = 61.3x (R2 = 0.9665), meaning that 61.3 mg N element was assimilated by 1.0 g dry biomass on average, which indicated that the nitrite transformation is a process of consuming nitrite as nitrogen source by Chlorella. The results demonstrated that the Chlorella suspension was able to assimilate nitrite efficiently, which implied the feasibility of using flue gas for mass production of Chlorella without preliminary removal of NOX.

Active Transport of Acidic Amino Acids in Suspension Cultured Brassica sp. Cells (배추과 식물현탁배양 세포내에서 산성 아미노산의 능동수송)

  • 조봉희
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.137-142
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    • 1995
  • The acidic amino acids, aspartate and glutamate, which have a negative charge in physiological pH, possess the same transport system as neutral amino acids according to the competitive inhibitory studies with the neutral amino acids. The neutral amino acids cotransported with one H+ per molecule, and one K+efflux per one molecule for charge compensation (Cho,1994), but the acidic amino acids cotransported with two H+ per one molecule, and one K+ efflux per one molecule. The active transport system, which possess the same carrier but cotransported with the different number of H+, reported for the first time. from the results, we can see that one of cotransported H+ protonated at first carboxyl group of pK$_3$ of acidic amino acids, and then as a neutral form cotransported with H+ Therefore, Brassica possess two amino acids transport system for 20 amino acids, namely general - and basic amino acids transport system. The evolutionary meaning of amino acid carriers described with other reported plants.

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Selection of Tropane Alkaloids High-Producing Lines by Single Cell Cloning of Hyoscyamus niger L. Root Cultures

  • Min, Ji-Yun;Park, Dong-Jin;Jeong, Mi-Jin;Song, Hyun-Jin;Kang, Seung-Mi;Kang, Young-Min;Choi, Myung-Suk
    • Journal of Korean Society of Forest Science
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    • v.98 no.2
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    • pp.142-147
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    • 2009
  • Hyoscyamus species is sources of the hypnotic and sedative drugs hyoscyamine and scopolamine. Single cells of Hyoscyamus niger were dissociated from suspension cultures and adventitious roots obtained from single-cell clones which were cultured on B5 medium containing 3% (w/v) sucrose, 0.1 mg/L IBA and 0.4% (w/v) gelrite. H. niger adventitious root lines showed wide variation in tropane alkaloids production and growth. An effective selection of 200 root lines was made possible by the application of the 'Dragendorff's reagent' for qualitative detection of the alkaloids from root. A high correlation coefficient (r=0.9390) was observed between the values obtained with the two methods based on HPLC and Dragendorff's reagent analysis. Among the selected roots, the highest scopolamine content was 16.64 mg/g DW (Hn-59), which was 8.82-fold more productive than the lowest alkaloid producing line (Hn-25). Here, we established an efficient selection method on tropane alkaloids production and suggest that the Dragendorff's reagent is of great practical value in selection of invisible compounds.

Taxol Production in Taxus Cell Cultures : 2. Effects of Sugar Concentration (주목 세포배양에 의한 Taxol 생산:2. 당농도의 영향)

  • 김진훈;박인석
    • KSBB Journal
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    • v.9 no.5
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    • pp.450-456
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    • 1994
  • The effects of sucrose on cell growth and anticancer drug taxol production were investigated in cell suspension cultures of Taxus cuspidata. The cells were cultured at various concentrations of sucrose (20 to $100g/\ell$). The highest specific growth rata was achieved at $40g/\ell$ of sucrose as 0.076 day-1 and the highest final cell density, 34 g DCW/$\ell$ after 25 days of culture, was obtained at $60g/\ell$ of sucrose. The cell yield(Yx/s) was found to be 0.55g cell/g sucrose and doubling time (Td) was 9.12 day. High concentration of sucrose (80, $100g/\ell$) and the addition of osmoticum enhanced the production of taxol significantly. The maximum taxol production was $1.36mg/\ell$ at $80g/\ell$ of sucrose, which was 0.01% as a specific content.

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Isolation and characterization of Auxin/indole-3-acetic acid 1 (Aux/IAA1) gene from poplar (Populus alba × P. glandulosa) (현사시나무에서 Auxin/indole-3-acetic acid 1 (Aux/IAA1) 유전자 분리 및 발현 특성 구명)

  • Bae, Eun-Kyung;Choi, Young-Im;Lee, Hyoshin;Choi, Ji Won
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.180-188
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    • 2019
  • Auxin plays a crucial regulatory role in plant growth and development processes. Three major classes of auxin-responsive transcription factors controlled by the Auxin/indole-3-acetic acid (Aux/IAA), Gretchen Hagen 3 (GH3), and small auxin up RNA (SAUR) genes regulate auxin signaling. Aux/IAA, in particular, encodes short-lived nuclear proteins that accumulate rapidly in response to auxin signaling. In this study, we isolated a PagAux/IAA1 gene from poplar (Populus alba ${\times}$ P. glandulosa) and investigated its expression characteristics. The PagAux/IAA1 cDNA codes for putative 200 amino acids polypeptide containing four conserved domains and two nuclear localization signals (NLSs). Utilizing Southern blot analysis, we confirmed that a single copy of the PagAux/IAA1 gene was present in the poplar genome. The expression of this gene is specific to leaves and flowers of the poplar. PagAux/IAA1 expressed in the early exponential growth phase of cell-cultured in suspension. PagAux/IAA1 expression level reduced in drought and salt stress conditions, and the presence of plant hormones such as abscisic acid. However, expression enhanced in cold stress, cambial cell division, and presence of plant hormones such as gibberellic acid and jasmonic acid. Thus, these results suggest that PagAux/IAA1 participates in cold stress response as well as developmental processes in the poplar.

Different Physiological Activity of Selected Rice Cultivars to Diphenylether Herbicide, Oxyfluorfen - I. Differential Responses of the Calli, Cells and Protoplasts (Oxyfluorfen에 대한 내성(耐性) 및 감수성(感受性) 벼품종(品種)의 생리활성(生理活性) 기구(機構) - I. Callus, 단세포(單細胞) 및 원형질체(原形質體) 반응(反應))

  • Kuk, Y.I.;Guh, J.O.;Lee, E.K.
    • Korean Journal of Weed Science
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    • v.16 no.1
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    • pp.42-53
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    • 1996
  • The tolerant and susceptible rice cultivars to oxyfluorfen were selected from 400 rice cultivars in a laboratory, and tested in comparison with barnyardgrass, a typical oxyfluorfen susceptible weed. The responses to oxyfluorfen in the different levels of calli, cells and protoplasts of the rice cultivars were investigated. $I_{50}$ value of the tolerant rice cultivars was about $10^{-4}M$, whereas that of the susceptible rice cultivars and barnyardgrass was about $10^{-6}M$, showing significant difference between the two groups. The growth rate of calli segregated from the tolerant rice cultivars was higher than that from the susceptible rice cultivars and barnyardgrass by treatment of oxyfluorfen to the calli. The growth rate of suspension-cultured cells of the tolerant rice cultivars was higher than that of the susceptible rice cultivars and barnyardgrass after treatment of oxyfluorfen. The viability of protoplasts from the tolerant rice cultivars was higher than that from the susceptible rice cultivars and barnyardgrass after one or two hours of oxyfluorfen treatment. The intactness of protoplasts from the tolerant rice cultivars was also higher than that form the susceptible rice cultivars and barnyardgrass.

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The effect of PDGF-BB loaded TCP/chitosan microgranules on new bone formation (혈소판유래성장인자를 함유한 TCP-chitosan 미세과립이 신생골조직 형성에 미치는 영향에 관한 연구)

  • Seol, Yang-Jo;Lee, Jue-Yeon;Kye, Seung-Beom;Lee, Young-Kyu;Kim, Won-Kyeong;Lee, Yong-Moo;Ku, Young;Han, Soo-Boo;Lee, Seung-Jin;Chung, Chong-Pyoung;Rhyu, In-Chul
    • Journal of Periodontal and Implant Science
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    • v.32 no.3
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    • pp.489-500
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    • 2002
  • The purpose of this study was to evaluate newly fabricated tricalcium phosphate(TCP)/chitosan microgranuls as bone substitutes. TCP/chitosan microgranules were fabricated by dropping TCP-chitosan suspension into the NaOH/ethanol solution. The size of microgranules could be controllable via airflow rate. PDGF-BB was loaded into the fabricated granules via freeze-drying methods(300 ng/20 mg). To evaluate cell proliferation, cultured osteoblasts cell lines(MC3T3-El) was dropped on the BioOss(R), chitosan microgranules, TCP/chitosan microgranules and cultured for 1, 7 , 14, and 28 days. Scanning electron microscopic observation was done after 7 days of culture and light microscopic examination was done after 28 days of culture. PDGF-BB release from the microgranules was tested. Rabbit calvarial defects(8 mm in diameter) were formed and chitosan, TCP/chitosan, PDGF-TCP/chitosan microgranules, and BioGran(R) were grafted to test the ability of new bone formation. At SEM view, the size of prepared microgranules was 250-1000 um and TCP powders were observed at the surface of TCP/chitosan microgranules. TCP powders gave roughness to the granules and this might help the attachment of osteoblasts. The pores formed between microgranules might be able to allow new bone ingrowth and vascularization. There were no significant differences in cell number among BioOss(R) and two microgranules at 28 day. Light and scanning electron microscopic examination showed that seeded osteoblastic cells were well attached to TCP/chitosan microgranules and proliferated in a multi-layer. PDGF-BB released from TCP/chitosan microgranules was at therapeutic concentration for at least 1 week. In rabbit calvarial defect models, PDGF-TCP/chitosan microgranules grafted sites showed thicker bone trabeculae pattern and faster bone maturation than others. These results suggested that the TCP/chitosan microgranules showed the potential as bone substitutes.

Characteristics and Pathogenicity of Hyphantria cunea Nuclear Polyhedrosis Virus (Hyphantria cunea Nuclear Polyhedrosis Virus의 특성 및 병원성)

  • Lee, Keun-Kwang;Kim, Myung-Kon;Park, Il-Woong
    • Applied Biological Chemistry
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    • v.38 no.5
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    • pp.435-439
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    • 1995
  • Some characteristics and pathogenicity of Hyphantria cunea nuclear polyhedrosis virus (HcNPV), a potential microbial pesticide was studied. H. cunea NPV replicated in the nucleus of S. frugiperda cells cultured in the TNMFH medium. In case of virus infected cell, prepolyhedra formation was observed at 24hrs post-infection. At 48 hrs post-infection, Most of the infected cell contained many mature polyhedra which were released into culture media 72 hrs post-infection, with the cells grown in suspension culture, pH of the culture medium increased during the virus replication: the pH of fresh medium was 6.35 and rose to 6.77 within 120 hrs. Polyhedra formed a band in linear density gradient of sucrose by centrifugation, which co-sedimented with $50{\sim}55%$ sucrose. The shape of the purified polyhedra was mostly tetragonal hexahedron and its size was about $2.5{\mu}m$. Electron microscopy and phase contrast microscopy showed that many bundled nucleocapsids were occluded in mature polyhedra at 48 hrs post infection. H. cunea larvae infected with NPV showed a higher motality in the second and third instar than in the fourth instar. Death rate of H. cunea larvae in the second and third instar fed with leaves coated with $1.5{\times}10^{9}{\sim}l.5{\times}10^{7}PIBs/ml$ reached more than 90%.

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A study of the effects of PDGF-BB on the characteristics of bone stromal and periodontal ligament cells (혈소판유래성장인자-BB가 골간질세포와 치주인대세포의 성상에 미치는 영향)

  • Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.949-965
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    • 1996
  • The main goal of periodontal therapy is to restore the lost periodontal tissue and establish the attachment appratus. Current acceptable therapeutic techniques are included : removal of diseased soft tissue, demineralization of exposed root surface, using the barrier membrane for preventing the downgrowth of gingival epithelial cell, insertion of graft materials as a scaffolding action, and biological mediators for promoting the cell activity. The latest concept one among them has been studied which based on the knowledge of cellular biology of destructed tissue. Platelet-derived growth factor(PDGF) is one of the polypeptide growth factor which have been reported as a biological mediator to regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the influences of the PDGF as biological mediator to periodontal ligament and bone marrow cell. Both right and left maxillary first molar were extracted from rat which had treated with 0.4% ${\beta}-Aminopropionitril$ for 5 days, and feeded until designed date to sacrifice under anesthesisa. Periodontal ligament were removed from the extracted socket of the rat, and cultured with Dulbecco's Modified Essential Medium(DMEM) contained with 10% Fetal Bovine Serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. Bone marrow cell were culture from bone marrow suspension with which washed out from femur with same medium. The study was performed to evaluate the effect of PDGF to periodontal ligament and bone cell, cell proliferation rate, total protein synthesis, and alkaline phosphatase activity of rat periodontal ligament(PDL) cell and bone stromal(RBS) cell in vitro. The effects of growth factors on both cells were measured at 3, 5th day after cell culture with (control group) or without growth factors(experimental group). The results were as follows: 1. The tendency of cell proliferation under the influence of PDGF showed more rapid proliferation pattern than control at 3 and 5 days after inoculation. 2. The activity of Alkaline phosphatase revealed 14, 80% increased respectively at 3, 5 days culture than control group. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(P<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P<0.05). 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group l(PDL 100%), 2(PDL 70%:GF 30%), and 3(PDL 50%:GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30%:GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05). From the above results, it is assumed that the PDGF on PDL cells and RMB cell culture. GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

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Isolation and Expression of Dormancy-associated protein 1 (DRM1) in Poplar (Populus alba × P. glandulosa) (현사시나무에서 Dormancy-associated protein 1 (DRM1) 유전자의 분리와 발현특성 구명)

  • Yoon, Seo-Kyung;Bae, Eun-Kyung;Choi, Hyunmo;Choi, Young-Im;Lee, Hyoshin
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.69-75
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    • 2017
  • Dormancy-associated protein (DRM) is involved in the dormancy physiology of plants and is conserved in almost all plant species. Recent studies found that DRM genes are involved in the abiotic stress response, and characterization studies of these genes have been conducted in several plants. However, few studies have focused on DRM genes in woody plants. Therefore, in this study, cDNA coding for DRM (PagDRM1) was isolated from poplar (Populus alba ${\times}$ P. glandulosa), and its structure and expression characteristics were investigated. PagDRM1 encodes a putative protein composed of 123 amino acids, and the protein contains two conserved domains (Domain I and Domain II). PagDRM1 is present as one or two copies in the poplar genome. Its expression level was highest in the stem, followed by mature leaves, roots, and flowers. During the growth of cultured cells in suspension, PagDRM1 was highly expressed from the late-exponential phase to the stationary phase. In addition, PagDRM1 expression increased in response to drought, salt stress, and treatment with plant hormones (e.g., abscisic acid and gibberellic acid). Therefore, we suggested that PagDRM1 not only plays an important role in the induction of dormancy, but also contributes to stress tolerance in plants.