• Title/Summary/Keyword: Cell suspension

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Synergistic Effect of Physico-chemical Treatment and Bacteriocin Produced by Enterococcus faecium MJ-14 (Enterococcus faecium MJ-14가 생산하는 박테리오신과 물리화학적 처리의 상승효과)

  • Lim Sung-Mee
    • Journal of Food Hygiene and Safety
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    • v.20 no.4
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    • pp.217-224
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    • 2005
  • When L. monocytogenes ($10^{5}CFU/mL$) at exponential phase cells were heated for 5 min at $65^{\circ}C$ in the presence of the bacteriocin (30 BU/mL) produced by E. faecium MJ-14, the number of viable cells was markedly reduced at p < 0.05. The bactericidal effect of bacteriocin showed synergism with combination ot organic acids (citric acid or acetic acid) or chemical preservatives (sodium benzoate, sodium lactate, sodium nitrate or potassium nitrate). For example, the number of viable cells was reduced by 4.8 log units under combination of the bacteriocin (30 BU/mL) and sodium nitrate ($100{\mu}g/mL$), while it was reduced by 1.1 log unit only under single treatment of the bacteriocin after 12 k at $37^{\circ}C$. The addition of the bacteriocin (300 BU/mL) into skim milk inoculated with L. monocytogenes ($10^{5}$CFU/mL) reduced the cells by 1.5 log unit, in case of the cell suspension stored at $4^{\circ}C$ for 24 hr. Moreover, L. monocytogenes was reduced by 2 log unit when stored at $-20^{\circ}C$ for 7 days in gound pork added with 300 BU/mL of 린e bacteriocin.

Ability for Biosynthesis of C27 Brassinosteroids by an Enzyme Pool Prepared from Cultured Cells of Phaseolus vulgaris (강낭콩 현탁배양세포에서 추출된 Enzyme Pool의 C27 Brassinosteroids 합성 능력)

  • Kim Tae-Wuk;Joo Se-Hwan;Kim Seong-Ki
    • Journal of Plant Biotechnology
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    • v.32 no.1
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    • pp.63-71
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    • 2005
  • A cell-free enzyme solution prepared from suspension cultured cells of Phaseolus vulgaris successfully mediated conversions of cholesterol $\to$ cholestanol and 6-deoxo-28-norteasterone $\leftrightarrow$ 6-deoxo-28-nor-3-dehydroteasterone $\leftrightarrow$ 6-deoxo-28-nortyphasterol $\to$ 6-deoxo-28-norcastasterone $\to$ 28-norcastasterone. Al-though conversion of cholestanol to 6-deoxo-28-norteasterone intermediated by 6-deoxo-28-norcathasterone was not demonstrated, this strongly suggests that a complete set of biosynthetic enzymes catalyzing reactions from cholesterol to 28-norcastasterone via 6-deoxo-28-nor type brassinosteroids is endogenously present in the cells, which demonstrates that a $C_{27}$ brassinosteroids biosynthetic pathway, namely the late C-6 oxidation for $C_{27}$ brassinosteroids, is operative in the cells. Additionally, the enzyme solution mediated conversion of 28-norcastasterone to castasterone in the presence of S-adenosyl-methionine and NADPH, providing that the $C_{27}$ brassinosteroids biosynthesis is an important route to generate castasterone in the cells. Together with our previous finding that castasterone can be biosynthesized by the same biosynthetic pathway in tomato, this study demonstrates that the $C_{27}$ brassinosteroids biosynthesis is a common alternative process to maintain endo-genous level of castasterone, an active $C_{28}$ brassinosteroid, in plants.

Quantitative Analysis of DNA Single-strand Breaks in EL 4 cells and Mouse Spleen Lymphocytes after Irradiation (방사선에 의한 EL 4 백서 백혈병 세포 및 정상 백서 비장 임파구 DNA Single-Strand Breaks의 정량적 분석과 측정)

  • Yoo Seong Yul;Cho Chul Koo;Koh Kyung Hwan;Park Woo Yoon;Park Young Hwan;Kim Sung Ho;Kim Tae Hwan;Chung In Yong
    • Radiation Oncology Journal
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    • v.8 no.2
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    • pp.137-144
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    • 1990
  • The filter elution technique was used to assay Co-60 $\gamma$ ray-induced DNA single-strand breaks(SSB) in EL 4 mouse leukemia cell and mouse spleen lymphocyte. The lymphocytes were stimulated with lipopolysaccharide (LPS, 20 $\mug/ml$) to label [${^3}H$] thymidine. EL 4 cells and lymphocytes in suspension were exposed at $0^{\circ}C$ to 0 Gy, 1 Gy, 5 Gy,10 Gy of Co-60 radiation and elution procedure was performed at PH 12.1. The number of DNA single-strand breaks increased with increasing doses of $\gamma$ rays. The strand scission factor (SSF) was estimated in each experiment (eluted volume 21 ml). The slope for EL 4 cells was $0.01301\pm0.00096\;Gy^{-1}(n=5)$ and the slope for lymphocytes was $0.01097\pm0.00091\;Gy^{-1}(n=5)$. The slopes were significantly different (P<0.005). Thus EL 4 cells were more sensitive to induction of DNA SSB by ionizing radiation than lymphocytes.

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Comparison of Effects of Different Activation Treatments on Development of Rabbit Embryos Reconstituted with Fetal Fibroblast

  • Lee, H.J.;Yoo, J.G.;Cho, S.R.;Lee, S.L.;Chong, J.R.;Yeo, H.J.;Hwang, J.M.;Park, J.S.;Yea, E.H.;Rho, G.J.;Choe, S.Y.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.22-22
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    • 2001
  • To produce reconstituted rabbit embryos with fetal fibroblasts, the present study was evaluated the efficiencies of the different fusion and activation conditions as assessments of subsequent development and chromosome in the embryos. New Zealand White rabbits were used throughout the study. Fetal fibroblasts collected from 22-d of fetuses were cultured in DMEM + 10% FBS in 5% $CO_2$ in air. The culture was maintained for 10 passages. In every passage half of cell suspension were kept In frozen. From rabbits treated with FSH in 30% PVP solution and hCG, oocytes were surgically collected from oviducts at 14 h post-hCG injection and stripped off their cumulus cells by re-pipetting in a 300 IU hyaluronidase solution. Oocytes with an extruded first polar body and dense cytoplasm were enucleated by micromanipulation in Ham's F-10 medium+7.5 g/$m\ell$ cytochalasin B. Euncleation was confirmed under a fluorescence microscope after staining with 5 g/$m\ell$ bisbenzimide for 2 min. Each enucleated oocyte was injected with a fetal fibroblast into a perivitelline space. Reconstructed eggs were compared fusion rates either at 2.0 ㎸/cm or 1.6 ㎸/cm(60 sec, double pulses). After fusion, all eggs were activated with the combination of 5 M ionomycin (5 min) and 10 g/$m\ell$ cycloheximide (CHX, 3h), and cultured in CRlaa medium and transferred into TCM199+10% FBS on day 3. Although there was not significantly differ in fusion rate between treatments (60%, 2.0 ㎸/cm vs. 79.4%, 1.6 ㎸/cm), none of them in the eggs fused with 2.0 ㎸/cm developed to blastocyst. In comparison of development and chromosome status between different activation treatments (Group 1; 5 M ionomycin/10 g/$m\ell$ CHX, Group 2; 5 M ionomycin/5 g/$m\ell$ CHX + 2 mM DMAP after fusion with 1.6 ㎸/cm), there were not differ in cleavage and development rates (67.3% and 28.9% in Group 1; 67% and 33% in Group 2). All out of 8 embryos evaluated in Group 1 appeared a normal diploid chromosome sets and mean number of cells (Mean SEM) on day 4.5 of culture was 141.5 23.15 (n=8). It can be concluded that the use of cycloheximide has not happened in chromosome abnormalities, and fetal fibroblasts can be used for cloning in rabbit.

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Antifungal Activity of Streptomyces pad anus isolate TH04 against Monilinia fructicola, Brown rot Fungus on Stone-fruits (잿빛무늬병균에 대한 Streptomyces padanus isolate TH04의 항균활성)

  • Lim, Tae-Heon;Choi, Yong-Hwa;Lee, Dong-Woon;Han, Sang-Sub;Cha, Byeong-Jin
    • The Korean Journal of Pesticide Science
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    • v.12 no.3
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    • pp.291-294
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    • 2008
  • The Streptomyces padanus isolate TH04, isolated from mummified peaches, showed strong antifungal activity to Monilinia fructicola. The inhibition activity of the isolate TH04 to mycelial growth and spore germination at 1% concentration of sub-antifungal powder made from culture suspension (CS) was ranged from 79.8% to 81.0% and from 73.9% to 75.8% to M. fructicola four strains, respectively. In the test of antifungal activity in mixed culture of the isolate and M. fructicola, inhibition rate was 7.5%, 86.8% and 94.0% in 0.01, 0.1, and 1% concentration of CS containing bacterial cell of the isolate, respectively. On apples (cultivar; Fuji), the control values of the isolate TH04 crude filtrates (0.1 and 1%) were 85.9% and 100%, respectively. The results suggest that the isolate TH04 indicate development possibility as biocontrol agent of brown rot caused by M. fructicola with the study on delivery method and fermentation condition to produce an antifungal compound.

Studies on the Pharmacodynamic Action of Methemoglobin (Methemoglobind의 약력학적(藥力學的) 작용(作用)에 관(關)한 연구(硏究))

  • Kim, Kwang-Yun
    • The Korean Journal of Pharmacology
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    • v.2 no.1 s.2
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    • pp.49-69
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    • 1966
  • For the purpose of stydying the pharmacodynamic action of methemoglobin, the author made the following experiments: 1. Preparation of hemoglobin and methemoglobin solutions: Red cell suspension from rabbit blood was hemolysed with distilled water and then divided into two portions. One portion was dialysed through cellophane paper and made isotonic with the proper amount of sodium chloride. The second portion was treated with sodium nitrite to convert hemoglobin to methemoglobin, dialysed through cellophane paper and made isotonic. 2. The concentration of methemoglobin in solution, plasma and urine was determined by Horecker and Brackette's method, and that of hemoglobin by the cyanmethemoglobin method. 3. The concentration of methemoglobin and hemoglobin in the plasma and urine of rabbits was measured at several intervals of time after infusion of the above samples. 4. The blood pressure and respiration of rabbits were recorded on a kymograph, and the effects of the samples on them were observed. 5. The effects of the samples on the movements of the in-situ heart and the isolated intestine of rabbits were studied. 6. The kidneys of rabbits were excised 4 to 5 hours after injection of the samples, and histopathological examinations were made. These experiments revealed the following results: 1. When methemoglobin solution was allowed to stand in room air, there was no decrease in the concentration of methemoglobin. 2. When methemoglobin solution was mixed with whole blood and incubated at $37^{\circ}C$, the concentration of methemoglobin decteased gradually. 3. After the infusion of methemoglobin and hemoglobin solutions, the rate of disappearance of methemoglobin in the plasma was more rapid than that of hemoglobin in the plasma. The higher the initial concentration in the plasma, the larger was the rate of disappearance of methemoglobin. 4. The rate of disappearance of methemoglobin was exceedingly rapid for 30 minutes after the infusion. 5. The urinary excretion of methemoglobin was more rapid than that of hemoglobin. 6. It would seem that the circulating blood contains substances which are promptly mobilized in the plasma to reduce methemoglobin to hemoglobin. 7. Moderate amounts of methemoglobin solution caused some rise in the blood pressure and a transient acceleration of the respiration of the rabbits. These effects of methemoglobin were milder than those of hemoglobin. 8. The movements of the in-situ heart and the isolated intestine of rabbits were accelerated by methemoglobin. These accelerating effects were milder than those of hemoglobin. 9. In the kidneys of rabbits treated with methemoglobin solution, hyperemia of the glomeruli, cloudy swelling and hemoglobin deposit in the tubular epithelium, hemoglobin casts in the tubular lumina of the proximal tubules, and interstitial congestion were constantly observed. There was no definite difference between the histological findings in the rabbit kidneys injected with methemoglobin, and those injected with hemoglobin solutions.

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Passive Immunity by Splenocyte Transfer against Amebic Meningoeneephalitis in Mice (세포에 의한 아메바성 수막뇌염에 대한 피동면역의 전달)

  • 임경일;유재숙
    • Parasites, Hosts and Diseases
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    • v.26 no.3
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    • pp.169-174
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    • 1988
  • The role of passive cell-mediated transfer of immunity against primary amoebic meningoen- cephalitis(PAME) in mice was studied. Waegleria fowleri, ITMAP 359, were cultured in CGVS medium. The ICR mice used were six week-old males of average weight of 15 g. Immunization was done by three intraperitoneal injections of $1{\times}10^6$ N. fowleri trophozoites at the interval of one week. Splenocytes were obtained from normal and immune mice spleens, and Ix107 cells were administered intraperitoneally into mice 3 days before challenge infection. Mice were infected intranasally with $7{\times}10^4$ N. fowleri trophozoites in a $3{\;}{\mu}l$ suspension under secobarbiturate anesthesia. Transplants of normal or immune splenocytes seem to alter the pattern of the PAME level- opment. The splenocytcs transferred from immune mice reduced the mortality rate in the JV. fowleri infected mice, as compared with the mice transferred with the same number of normal splenocytes or without splenocyte, The blastogenic response of the splenocytes to both lipopoly- saccharide and concanavalin A was elevated on duty 7 after infection the mice transinoculated with immune splenocytes. The serum antibody titers in the mice transferred with immune spleno- cytes were increased gradually from day 7 up to day 20 after infections by mean of ELISA. It is suggested that the transfer of splenocytes from immuniged mice conferred immunity against N. fowleri infection.

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Plant Production from Desiccated Somatic Embryos of Acanthopanax chiisanensis (지리오가피 (Acanthopanax chiisanensis) 체세포배의 건조처리를 통한 식물체 증식)

  • Lee, Kang-Seop;Bang, Keuk-Soo;Choi, Yong-Eui;Ahn, Byung-Yong
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.381-385
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    • 2003
  • An efficient method of plant regeneration from Acanthopanax chiisanensis somatic embryos was developed. Cotyledonary somatic embryos were obtained in liquid Murashige and Skoog (MS) medium from embryogenic cell suspension cultures. They were desiccated for 0 to 72 hr and then cultured on MS medium containing NAA, BA, GA$_3$, (0-0.5mg/L). The highest multiple shoots formation (100%) was obtained from 72 hr desiccated somatic embryos on ifs medium with 0.5mg/L NAA+0.5mg/L BA or 0.5 mg/L NAA+0.5mg/L BA+0.5mg/L GA$_3$ after 6 weeks culture. Plant conversion from multiple shoots was not high. The highest plant conversion from multiple shoots was obtained on 1/3MS medium with 1.0mg/L GA$_3$. Converted plantlets were transferred to ex vitro condition and the highest survival rate (70%) of the plantlets was obtained on plastic pots containing vermiculite and sand. These results indicate that micropropagation procedure can be applied for an efficient mass propagation of Acanthopanax chiisanensis.

Effect of kinds and concentrations of osmoticum on somatic embryo induction and germination from suspended embryogenic cell in Larix kaempferi (낙엽송(Larix kaempferi) 현탁배양된 배발생세포로부터 체세포배 유도 및 발아를 위한 삼투압제 종류 및 농도 효과)

  • Kim, Yong-Wook
    • Journal of Plant Biotechnology
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    • v.40 no.3
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    • pp.141-146
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    • 2013
  • This study was conducted to examine suspended embryogenic cells growth with days of culture, effects of various kinds/concentrations of osmoticum for induction of somatic embryos (SEs), following somatic embryos germination or plantlet regeneration. The proliferation pattern of embryogenic cells in suspension culture is characterized by settled cells volume (SCV) increased with the duration of culture with marked the maxium of SCV (10.1 ml) in 18 days of culture, however the SCV of cells gradually decreased after that. In comparison of kinds/concentrations of osmoticum on somatic embryo induction, the highest induction number (352.3/g FW) of the SE was showed in 0.2 M sucrose, in addition, we also observed some effects with treatments of 0.2 M maltose (203.7) and 0.3 M maltose (193.7), respectively. However, no somatic embryos produced in treatments of 7.5% PEG plus 0.15 M sucrose or maltose. In comparison of germination efficiency of SEs which occurred from the treatments of various kinds/ concentrations of osmoticum, the highest induction frequency of cotyledon (25.2%) was obtained from SEs that produced 0.3 M maltose, however, the best occurrence rates of hypocotyl (39%), radicle (30.3%) and plantlet regeneration (3.5%) were observed from the 0.2 M sucrose treatment, respectively.

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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