• Title/Summary/Keyword: Cell suspension

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Improvement of an hydroxyapatite bead adherence assay for streptococcus sanguis (Streptococcus sanguis의 구형 Hydroxyapatite 비드에의 부착 Assay 방법의 개량)

  • 최선진;이시영;송요한
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.48-55
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    • 1989
  • The purpose of the present investigation was to improve several procedures being used in the adherence assay of Streptococcus sanguis cells to hydroxyapatite (HA) beads and to study the effect of the beads on the counting of radioactivity. The standard adhere assay involved the adherence of radiolabeled bacteria to 40mg of HA beads. The beads were mixed with ['H]thymidine-labeled bacterial cells and incubated for 60 minutes at room temperature. Unadsorbed cells were removed, the beads with adsorbed cells were dried, and the radioactivity was monitered in a scintillation spectrometer. The 30 seconds sonication of cells in a form of long chains appeared to be adequate for obtaining mostly singlet or doublet cells. Unlike the counting of S. sanguis cell suspension, bacterial cells adhered to HA or saliva-coated HA(SHA) required smaller volume (2.5ml) of scintillaton fluid for better counting. Eighteen percent quenching of counts could be attributed to the beads. Among 3 procedures commonly used to equilibrate the beads for adherence assay, no differences were found in their effectiveness. The HA beads on which the bacteria remained attached in scintillant during the counting were found to be the source of sample self-absorption representing 34.5% of the total radioactivity counts resulting from the beads dissolved in HCl solution.

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Influence of Exposure to Extremely Low Frequency Magnetic Field on Neuroendocrine Cells and Hormones in Stomach of Rats

  • Hong, Min-Eui;Yoon, Kyu-Hyun;Jung, Yoon-Yang;Lee, Tae-Jin;Park, Eon-Sub;Sohn, Uy-Dong;Jeong, Ji-Hoon
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.3
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    • pp.137-142
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    • 2011
  • Extremely low frequency magnetic fields (ELF-MF) have the ability to produce a variety of behavioral and physiological changes in animals. The stomach, as the most sensitive part of the neuroendocrine organ of the gastrointestinal tract, is crucial for the initiation of a full stress response against all harmful stress. Thus, the purpose of this study was to examine whether ELF-MF stimuli induce changes in the activity of neuroendocrine cells, considering their involvement in endocrine or paracrine effect on surrounding cells. The exposure to ELF-MF (durations of 24 h and 1 or 2 weeks, 60 Hz frequency, 0.1 mT intensity) altered the distribution and occurrence of gastrin, ghrelin and somatostatinpositive endocrine cells in the stomach of rats. The change, however, in the secretion of those hormones into blood from endocrine cells did not appear significantly with ELF-MF exposure. Comparing with sham control, ELF-MF exposure for 1 and 2 week induced an increase in $BaSO_4$ suspension propelling ratio of gastrointestinal tract, indicating that ELF-MF affects gastrointestinal motility. Our study revealed that ELF-MF exposure might influence the activity of endocrine cells, an important element of the intrinsic regulatory system in the digestive tract. The pathophysiological character of these changes and the mechanism responsible for neuroendocrine cell are still unclear and require further studies.

Relationship of Cotyledon Number with Procambium Differentiation in Somatic Embryogenesis of Codonopsis lanceolata L. (더덕의 체세포배발생에서 전형층분화와 자엽수의 관계)

  • Choi Pil-Son;Soh Woong-Young;Cho Duck-Yee;Liu Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.135-138
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    • 2005
  • Embryogenic callus was obtained from cotyledonary explants of Codonopsis lanceloata on Murashige & Skoog's medium supplemented with 1 mg/L 2,4-D. Suspension cultures of the embryogenic calli were grown on a shaker at 100 strokes/min, and then the calli were subcultured for 2 weeks in 2,4-D-free medium to produce somatic embryo. In somatic embryos at the globular stage, cotyledon initials began to differentiate themselves in the near distal end of the procambial strand. Dicotyledons, tricotyledon, tetracotyledon and fused cotyledon were differentiated from the distal ends of two, three, four and circular procambial strands, respectively. Nearly circular procambial strand in lower hypocotyls were independently differentiated into two, three, four procambial tissues at cotyledonary node and cotyledons to form somatic embryos with dicotyledon, tricotyledon, tetracotyledon. If the distal subepidermal cells of globular embryo exclusively became cotyledon initials, the torpedo or cotyledonary embryo was characterized by somatic embryos with fused cotyledon.

Taxol Production in Taxus sap. Cell Cultures 1. Studies on Taxol Content in Yew Trees and Cultured Plant Cells (주목 세포배양에 의한 Taxoll 생산 1.주목 부위 및 서식 고도별 Taxol 함량 및 세포주 유도에 관한 연구)

  • 강인선;전정욱
    • KSBB Journal
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    • v.9 no.3
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    • pp.299-305
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    • 1994
  • Taxol contents in various parts of 15 years old yew tree were determined. The descending order of taxol content per unit mass was stem bark, root bark, needle and seed. In the seed, that order was seed coat, embryo and endosperm. The total amount of taxol extractable from a 15 years old yew tree was 1.68 gram. This amount was distributed in needle, stem bark, root bark and seeds as 48.0, 23.8, 27.9 and 0.4%, respectively. Altitudinal variation of taxol content was also observed. More taxol was observed in yew trees grown at high altitude over 1000m above sea level. Calli and suspension cultures were induced from various yew trees. The presence of taxo] in cultured cells was established bv HPLC. The taxol content in cultured cells were different according to the source of explants. These results may be useful for the goal of large scale taxol production by cultured yew tree cells.

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Selective Detection of Campylobacter sp. and Campylobacter jejuni in Meat Food by Polymerase Chain Reaction (PCR을 이용한 육류 내 Campylobacter sp. 및 Campylobacter jejuni의 분리 검출)

  • Joo, Jong-Won;Hong, Kyung-Pyo;Kim, Yong-Hui;Cho, Sang-Buem
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.5
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    • pp.753-759
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    • 2008
  • The principal objective of this study was to develop the optimum oligonucleotide primers for the simple detection of Campylobacter in food samples. In order to achieve this goal, a variety of oligonucleotide primers were designed via the modification of 16S rDNA, ceuE and mapA sequences of Campylobacter. Through the subsequent analysis of the specificity and sensitivity of primers, two types of oligonucleotide primers, CB4 and CJ1, were selected for Campylobacter genus-specific and C. jejuni species-specific primers, respectively. The detection limit was found to be $10^0{\sim}10^1$ cells per reaction with the prepared cell suspension, however, the sensitivity in the meat samples was less, at $10^1{\sim}10^2$. We suggested that PCR inhibitors such as hemoglobin or immunoglobulin in pork or beef influenced.

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Production of Chiral Phenyl Oxirane by Rhodosporidium toruloides in Hollow-fiber Reactor (Rhodosporidium toruloides를 이용한 Hollow-fiber 반응기에서의 광학활성 Phenyl Oxirane 생산)

  • 김희숙;박성훈;이은열
    • Journal of Life Science
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    • v.13 no.6
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    • pp.788-793
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    • 2003
  • Production of chiral phenyl oxirane by Rhodosporidium toruloides SJ-4 was investigated. Racemic phenyl oxirane was kinetically resolved by enantioselective hydrolysis reaction by epoxide hydrolase of R. toruloides in two-phase hollow-fiber reactor system. Dodecane with high concentration of the racemic substrate passed through the lumen side and cell suspension was recirculated through the shell side of the hollow fiber reactor For the removal of phenyl-1, 2-ethandiol to reduce the product inhibition to biocatalysts, another hollow-fiber reactor was employed to extract the diol. Racemic phenyl oxirane up to 300 mM was enantioselectively resolved with high enantiopurity (>99% ee) in hollow-fiber reactor system.

Ethanol changes atpB gene expression and proton permeability in Streptococcus mutans (에탄올이 Streptococcus mutans의 atpB 유전자 발현 및 양성자 투과성에 미치는 영향)

  • Cho, Chul Min;Park, Yong Jin;Lee, Sae A;Kim, Jin Bom;Kang, Jung Sook
    • Journal of Korean Academy of Oral Health
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    • v.42 no.4
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    • pp.224-228
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    • 2018
  • Objectives: As a first step to study the anticaries effect of ethanol alone, we investigated the effects of ethanol on the expression levels of the atpB gene and proton permeability of Streptococcus mutans in suspension cultures. Methods: S. mutans UA159 was grown in brain heart infusion medium at either pH 4.8 or 6.8. The total extracted RNA was reverse-transcribed into cDNA using a $Superscript^{TM}$ First-Strand Synthesis System. The resulting cDNA and negative controls were amplified by ABI PRISM 7700 real-time PCR system with SYBR Green PCR Master Mix. For proton flux assay, bacterial suspensions were titrated to pH 4.6 with 0.5 M HCl, and then additional 0.5 M HCl was added to decrease the pH values by approximately 0.4 units. The subsequent increase in pH was monitored using a glass electrode. Ten percent (v/v) butanol was added to the suspensions at 80 min to disrupt the cell membrane. Results: In a concentration-dependent manner, ethanol alone not only decreased the growth rate of S. mutans and the expression of the atpB gene but also increased the proton permeability at both pH 4.8 and 6.8. Conclusions: These findings suggest that ethanol has the potential for an anticaries ingredient. We believe that ethanol may be used together with fluoride and/or other cariostatic agents in order to develop better anticaries toothpastes and/or mouthrinses.

Evaluation of SERS Nanoparticles to Detect Bacillus cereus and Bacillus thuringiensis

  • Hong, Jeehwa;Qin, Jianwei;Van Kessel, Jo Ann S.;Oh, Mirae;Dhakal, Sagar;Lee, Hoonsoo;Hwang, Chansong;Chan, Diane E.;Kim, Dongho;Cho, Hyunjeong;Kim, Moon S.
    • Journal of Biosystems Engineering
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    • v.43 no.4
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    • pp.394-400
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    • 2018
  • Purpose: This research evaluated five types of nanoparticles to develop a surface-enhanced Raman spectroscopy (SERS) method for the rapid detection of two Bacillus species (Bacillus cereus and Bacillus thuringiensis) that are commonly found on fresh produce, which can cause food poisoning. Methods: Bacterial concentrations were adjusted to a constant turbidity, and a total of $30{\mu}L$ of each Bacillus cell suspension was prepared for each nanoparticle. A point-scan Raman system with laser light source of wavelength 785 nm was used to obtain SERS data. Results: There was no qualitative difference in the SERS data of B. cereus and B. thuringiensis for any of the five nanoparticles. Three gold nanoparticles, stabilized in either citrate buffer or ethanol, showed subtle differences in Raman intensities of two Bacillus species at $877.7cm^{-1}$. Conclusions: Among the three types of nanoparticles, the gold nanoparticles stabilized in citrate buffer showed the lowest standard deviation, followed by gold nanoparticles stabilized in ethanol. This result supports the potential application of gold nanoparticles for SERS-based detection of B. cereus and B. thuringiensis.

Scutellaria baicalensis Inhibits Coxsackievirus B3-Induced Myocarditis Via AKT and p38 Pathways

  • Fu, Qiang;Gao, Lu;Fu, Xiao;Meng, Qinghua;Lu, Zhihong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1230-1239
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    • 2019
  • Scutellaria baicalensis Georgi has been widely used in China for treatment of various diseases. This study investigated the effect of Scutellaria baicalensis Georgi extracts (SBE) against Coxsackievirus B3 (CVB3)-induced myocarditis in vitro and in vivo. In vitro, Hela cells and primary myocardial cells were infected with CVB3 and treated with SBE ($50-800{\mu}g/ml$) and ribavirin ($200{\mu}M$) for 48 h and then determined by CCK8 assay. Real-time PCR and western blotting assays were performed. In vivo, a myocarditis model was induced in male BALB/c mice by injecting CVB3 suspension intraperitoneally for three times, followed by treatment with SBE (400 and 200 mg/kg) and ribavirin (100 mg/kg) for 28 days. SBE ameliorated the cytotoxicity of CVB3 in Hela cells, especially at $400{\mu}g/ml$ (39.93% vs 65.67%, p < 0.05) without influencing cell growth and also significantly reduced CVB3 replication in primary myocardial cells. The levels of AKT, ERK, and p38 were increased after CVB3 infection. SBE could downregulate the expressions of AKT and p38. In vivo, the mortality rate from CVB3 reached to 66.67%, while 10.00% and 23.33% of this came after 400 and 200 mg/kg SBE treatment, respectively (p < 0.05). The CVB3 replication was obviously reduced after SBE administration from day 5. Similarly, the levels of AKT, ERK, and p38 mRNAs and proteins were increased, and SBE suppressed the expression of AKT and p38. Our study indicates that SBE is a promising potent antiviral agent against CVB3-induced myocarditis by inhibition of virus replication via depressing AKT and p38 expressions.

Architectural Design Related to the Growth Principles of Tree (나무의 성장원리를 응용한 건축 디자인에 관한 연구)

  • Kim, Tai Young
    • The Journal of the Convergence on Culture Technology
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    • v.7 no.1
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    • pp.49-57
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    • 2021
  • This study is to find ways to self-produce energy even in buildings through the system of trees that grow by themselves focused on literatures and case studies. It is divided into the structure, circulation and reaction system of tree. 1) In the structural system, the tree is divided into the shoot and root system, and maintains rigidity with the cell membranes. The wind resistance caused by the trunk and crown can be applied to the seismic structure principle of building, and the role of platelike buttresses of lateral roots can be applied to the horizontal truss and suspension bridge. 2) In the circulation system, the transpiration action through the fine stomata of the leaves can be a very effective cooling means because a large amount of heat is released and this method can be directly introduced into the cooling of buildings. 3) In the responsive system, the response system according to environmental changes that can be read from the leaves and flowers of trees can be applied to the roof and exterior design of buildings through the use of new sensing technologies and materials.