• Title/Summary/Keyword: doubling condition

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The importance of post-thaw subculture for standardizing cellular activity of fresh or cryopreserved mouse embryonic stem cells

  • Ko, Dong Woo;Yoon, Jung Ki;Ahn, Jong il;Lee, Myungook;Yang, Woo Sub;Ahn, Ji Yeon;Lim, Jeong Mook
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.3
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    • pp.335-343
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    • 2018
  • Objective: Remarkable difference in cellular activity was found between early and late subpassaged embryonic stem cell (ESCs) lines, which can be created by subtle changes in cell manipulation protocol. This study subsequently examined whether post-thaw subculture of early subpassaged ESC lines could further affect the activity of the ESCs. Methods: Fresh (as a control treatment) or cryopreserved F1 hybrid (B6CBAF1) early ESC lines (C57BL/6xCBA) of the 4 (P4) or the 19 passage (P19) were subcultured once, twice or six times under the same condition. The post-thaw survival of the ESCs was monitored after the post-treatment subculture and the ability of cell proliferation, reactive oxygen species (ROS) generation, apoptosis and mitochondrial ATP synthesis was subsequently examined. Results: Regardless of the subculture number, P19 ESCs showed better (p<0.05) doubling time and less ATP production than P4 ESCs and such difference was not influenced by fresh or cryopreservation. The difference between P4 and P19 ESC lines became decreased as the post-treatment subculture was increased and the six times subculture eliminated such difference. Similarly, transient but prominent difference in ROS production and apoptotic cell number was detected between P4 and P19 ESCs only at the 1st subculture after treatment, but no statistical differences between two ESC lines was detected in other observations. Conclusion: The results of this study suggest that post-thaw subculture of ESCs under the same environment is recommended for standardizing their cellular activity. The activity of cell proliferation ability and ATP synthesis can be used as parameters for quality control of ESCs.

Inhibitory Effect of Petroleum Ether Extract of Panaz Ginseng Root against Growth of Human Cancer Cells (고려인삼 중 Petroleum Ether 추출물의 인체 암세포 증식억제효과)

  • 이선희;황우익
    • Journal of Ginseng Research
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    • v.10 no.2
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    • pp.141-150
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    • 1986
  • This study was attempted to screen the cytotoxic activity of petroleum ether ex- tract from panax ginseng root against human colon cancer cells. Two extracts of panax ginseng root, crude and partially purified, were used for this experiment. The crude extract was prepared by extraction with petroleum ether using Soxhlet aparatus for 12 to 15 hours from panax ginseng and the extract was partially purified by silicic acid column with mixture of petroleum ether: ethyl ether (70 : 30, v/v). Three species of human colon cancer cells, HRT-18, HCT-48 and HT-29, were maintained in DMEM (Dulbecco's modified Eagle medium), and the cells were cultured in DMEM containing serial concentration of the crude or partially purified fraction to observe the cytotoxic activity of the both extracts. The effects of incubation time and concentration of the both extracts in culture medium against the growth of the each cancer cell were determined. The results obtained are summarized as follows: 1. The doubling times of the HRT-18, HCT-48 and HT-29 cells were about 20, 24 and 22 hours, respectively. 2, The inhibitory effects of the crude extract on the growth of cancer cells were increased according to the rise of concentration of the extract and incubation time. 3. The inhibitory effect of partially purified fraction on the growth of HRT-18 cell was about 4 times stronger than that of the crude extract under same experimental condition. 4 The inhibitory effects of the crude and purified fraction on the growth of each cancer cell were shown difference by the kind of the cancer cell. In view of the above results, it could be said that the petroleum ether extract of panax ginseng root inhibited the division of the human colon cancer cell, in vitro.

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Studies on the Petroleum hydrocarbon-utilizing Micro-organisms(Part 2) - On the Production of Single Cell Protein from Petroleum hydrocarbon with a yeast strain - (석유 탄화수소 이용 미생물에 관한 연구 (제 2 보) - 효모를 이용한 석유탄화수소로 부터 단백질 생산에 관하여 -)

  • Lee, Ke-Ho;Shin, Hyun-Kyung
    • Applied Biological Chemistry
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    • v.14 no.1
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    • pp.9-18
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    • 1971
  • In order to obtain basic information on the production of single cell protein from petroleum, more than 400 yeast strains were isolated from various soil samples in Korea utilizing petroleum hydrocarbon as the sole carbon source. A yeast strain showing the highest cell yield among the isolated strains was selected and identified. The optimal culture condition was searched in the flasks shaken throughout the procedure. And the growing characteristics for the selected yeast strain and chemical analysis of the yeast cell component were carried out. The results obtained were as follows: 1. The selected yeast strain was identified as Candida curvata and we named it Candida curvata-SNU 70. 2. The composition of the medium proposed for the present yeast strain is: Light Gas Oil 30ml, Urea 400mg, Ammonium sulfate 100mg, Potasium phosphate (monobasic) 670mg, Sodium phosphate (dibasic) 330mg, Magnesium sulfate 500mg, Calcium carbonate 3g, Yeast extract 50mg, Tween 20 0.05ml, Tap water 1,000ml. 3. Other culture conditions employed for the yeast were pH 5.5-7.0, temp. $30^{\circ}C$ under an affluent aerobic state. 4. Addition of light gas oil in portions to the culture media as the growth proceeded was more effective, especially in the cultivation on the higher oil concentration media. 5. Studies on the propagation of the yeast cells in the light gas oil medium revealed that the yeast has the lag phase lasted 16 hours and the logarithmic growth phase covered 16 to 28 hours. The specific growth rate was about $0.22\;hr^{-1}$ and doubling time was 3.2 hrs. during the logarithmic growth phase. 6. Under the cultural condition employed, the cell yield against the amount of light gas oil (wt%) was 16.1% and the protein content of the dried yeast cells was 48.4%.

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A Study on the Production of Yeast Utilizing Ethanol as a Sole Carbon Source (Ethanol 이용 미생물에 의한 단세포 단백질 생산에 관한연구)

  • Lee, Ke-Ho;Ha, Jin-Hong
    • Applied Biological Chemistry
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    • v.16 no.1
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    • pp.1-11
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    • 1973
  • In order to obtain the basic informations on the production of single cell protein from ethanol, 145 yeast strains utilizing ethanol as a sole carbon source were isolated from 32 soil samples in Korea. A yeast strain showing the highest cell yield among the isolated strains was selected and identified. The optimum culture condition, utilization of other carbon sources and the cultural characteristics for the selected yeast, and the chemical analysis of the yeast cell composition, and utilization of ethanol by the selected yeast were investigated. All the culture was carried out in the shaking flasks. The results obtained were as follows: 1. The selected yeast strain was identified as Debaryomyces nicotianae-SNU 72. 2. The optimum composition of the medium for the selected yeast is : Ethanol 40 ml, Urea 0.5 g, Potassium phosphate (dibasic) 0.5 g, Ammoium phosphate (monobasic) 0.15 g, Magnesium sulfate 0.05 g, Calcium chloride 0.01g, Yeast extract 0.005 g, Tap water 1000 ml. 3. The optimum pH was 5.0-5.5, the optimum temperature $30-33^{\circ}C$ and the aerobic state was unimportant. 4. Utilization of methanol, n-propanol, iso-propanol, n-butanol, iso-butanol, tert-amyl alcohol and acetic acid by the selected yeast was very weak. So substitution of the subtrate was thought to be impossible. 5. Studies on the propagation of the yeast cells showed that the lag phase of the yeast cells lasted 16 hours, and the logarithmic growth phase extended 16 to 28 hours. The specific growth rate was about $0.19\;hr^{-1}$ and the doubling time was 3.6 hours during the logarithmic growth phase. 6. As the result of the chemical analysis of the dry yeast cells, the content rate of the crude protein was 55.19 %, the content of others was similar to the average content of the yeast component. 7. After 34 hours cultivation, under the optimum culture condition investigated, the dry cell yield against the amount of the added ethanol was 53.4 % (W/V%), the dry cell yield against the amount of the utilized ethanol was 73.6 % (W/V%), the evaporation rate of ethanol was about 19.1 %.

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Effect of Nutrients on Competition among the Harmful Dinoflagellates Cochlodinium polykrikoides and the Diatom Skeletonema sp. in Jaran Bay Using a Mathematical Model (수치모델을 활용한 자란만에서 유해 와편모조류 Cochlodinium polykrikoides와 규조류 Skeletonema sp.의 경쟁에 미치는 영양염의 영향)

  • Oh, Seok Jin;Kim, Hyun Jung;Kwon, Hyeong Kyu;Yang, Han-Soeb;Kim, Seok Yoon
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.20 no.2
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    • pp.92-101
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    • 2015
  • We investigated cause of non-outbreak of Cochlodinium polykrikoides blooms in the western coast of Jaran Bay during summer, 2013, based on the species competition among C. polykrikoides and Skeletonema sp. using a mathematical model. As a result of the model simulation where the nutrient conditions in Jaran Bay was applied during summer, the cell density of Skeletonema sp. was higher than that of C. polykrikoides. In the sensitivity analyses by doubling and halving the parameters, any parameter had little effect on the cell density of C. polykrikoides. The cell density of Skeletonema sp. was significantly affected by changes in the parameter values. These results indicated that the growth of C. polykrikoides could be unaffected by rapidly changing environments. However, the growth of Skeletonema sp. may have been promoted by the changing nutrient supply of coast environments. Therefore, C. polykrikoides might have been suppressed by diatom blooms, such as Skeletonema sp., in changing nutrient supply condition of Jaran Bay.

Frequency Doubling in LiIO3 Crystals by the Ring Enhancement Cavity (고리형 증폭 공진기에 의한 LiIO3결정에서 제2조화파 발생)

  • Kim, Sang-Gee
    • Journal of Korean Ophthalmic Optics Society
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    • v.4 no.2
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    • pp.45-49
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    • 1999
  • The second harmonic, wavelength is 397nm, of the continuous wave diode laser, whose maximum power is 35mW, was generated in $LiIO_3$ crystals in a ring enhancement cavity. 5mm- and 10mm-long crystals cut $43.21^{\circ}$ for optic axis were used in this experiment. Both surfaces of those were anti-reflection coated for 794nm. In case the crystal was inserted into the cavity, the condition of separation between two concave mirrors for the optimum mode matching was found. The conversion efficiency of second harmonic generation was increased by the resonant enhancement of pumping power in the ring enhancement cavity, and the frequency of diode laser was locked to that of the counter-propagation mode generated from the surface of crystal. When the pumping power was 28 mW, the infrared buildup factor was about 45 without the crystal, and 14 with the crystal due to the transmission loss of crystal. The maximum second harmonic powers of $1.5{\mu}W$ and $6.6{\mu}W$ were obtained, and corresponding conversion efficiencies were $(6.584{\pm}0.56){\times}10^{-3}$%, $2.6{\pm}0.21){\times}10%{-2}$% in 5mm- and 10mm-long $LiIO_3$, respectively.

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Long-term Effects of Change in Family Structure On Achievement During Transition to Adulthood : Focusing on the effect of parental divorce/death on health condition, depression and educational attainment (가족구조의 변화가 성인이행기 발달에 미치는 영향 : 주관적 건강상태, 우울, 교육성취를 중심으로)

  • Kim, Yeonwoo
    • Korean Journal of Social Welfare Studies
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    • v.41 no.4
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    • pp.225-246
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    • 2010
  • The family environment children are exposed to growing up greatly influences their future potential and achievements. Previous findings show that changes in family structure during childhood, particularly those resulting from divorce or death, cause lasting negative consequence that affect the child physically, psychologically, economically, and socially. Unfortunately, single-parent households are becoming increasingly common in Korea, nearly doubling to more than a million cases in the last two decades. Existing domestic and international studies of this area tend to focus on the short-term effects of growing up in a single-parent household. In addition, these studies group their samples in ways that result in findings that may be too broad or are not necessarily an accurate representation of the subjects. This study attempts to address some of these shortcomings by focusing on the long-term effects of how changes in family structure early in children's lives affect achievement during their transition to adulthood. In addition, it takes into account the development cycle the child is in at the time of family restructuring, and what kind of long-term effects result from that. In this analysis, we find that there are several cases of statistically significantly differences in domain achievement depending on the developmental stage the child was in when the parental divorce or death occurred. The findings indicate that changes in family structure during the infant/toddler period influence health condition and depression, while changes in family structure during middle-childhood and adolescence do not. Meanwhile, changes in family structure during any point in the developmental stages have negative effects on educational attainment, with the severity of these negative effects depending on when the family changes occur. The negative effect on educational attainment is most prominent when a change in family structure occurs during the infant/toddler period, followed by adolescence, then middle-childhood.