• Title/Summary/Keyword: cell growth yield

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Regulatory Mechanism of Radiation-induced Cancer Cell Death by the Change of Cell Cycle (세포주기 변화에 타른 방사선 유도 암세포 사망의 조절기전)

  • Jeong Soo-Jin;Jeong Min-Ho;Jang Ji-Yeon;Jo Wol-Soon;Nam Byung-Hyouk;Jeong Min-Za;Lim Young-Jin;Jang Byung Gon;Youn Seon-Min;Lee Hyung Sik;Hur Won Joo;Yang Kwang Mo
    • Radiation Oncology Journal
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    • v.21 no.4
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    • pp.306-314
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    • 2003
  • Purpose : In our Previous study, we have shown the main cel1 death pattern Induced by irradiation or protein tyrosine kinase (PTK) inhibitors in K562 human myeiogenous leukemic cell line. Death of the cells treated with irradiation alone was characterized by mitotic catastrophe and typical radiation-induced apoptosis was accelerated by herblmycin A (HMA). Both types of cell death were inhibited by genistein. In this study, we investigated the effects of HMA and genistein on cell cycle regulation and its correlation with the alterations of radiation-induced cell death. Materials and Methods: K562 cells In exponential growth phase were used for this study. The cells were Irradiated with 10 Gy using 6 MeV Linac (200-300 cGy/min). Immediately after irradiation, cells were treated with 250 nM of HMA or 25 $\mu$N of genistein. The distributions of cell cycle, the expressions of cell cycle-related protein, the activities of cyclin-dependent kinase, and the yield of senescence and differentiation were analyzed. Results: X-irradiated cells were arrested In the G2 phase of the cell cycle but unlike the p53-positive cells, they were not able to sustain the cell cycle arrest. An accumulation of cells in G2 phase of first ceil-cycle post-treatment and an increase of cyclin Bl were correlated with spontaneous, premature, chromosome condensation and mitotic catastrophe. HMA induced rapid G2 checkpoint abrogation and concomitant p53-independent Gl accumulation. HMA-induced cell cycle modifications correlated with the increase of CDK2 kinase activity, the decrease of the expressions of cyclins I and A and of CDK2 kinase activity, and the enhancement of radiation-induced apoptosis. Genistein maintained cells that were arrested in the G2-phase, decreased the expressions of cyclin Bl and cdc25c and cdc25C kinase activity, increased the expression of pl6, and sustained senescence and megakaryocytic differentiation. Conclusion: The effects of HMA and genistein on the radiation-induced cell death of KS62 cells were closely related to the cell cycle regulatory activities. In this study, we present a unique and reproducible model in which for investigating the mechanisms of various, radiation-induced, cancer cell death patterns. Further evaluation by using this model will provide a potent target for a new strategy of radiotherapy.

Bioethanol Production from Seaweed Kappaphycus alvarezii by Simultaneous Saccharification and Fermentation (홍조류(Kappaphycus alvarezii)의 동시 당화 발효를 이용한 바이오에탄올의 생산)

  • Ra, Chae Hun;Kim, Sung-Koo
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.145-149
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    • 2016
  • Thermal acid hydrolysis pretreatment of Kappaphycus alvarezii was carried out with 12% (w/v) seaweed slurry and 180 mM H2SO4 at 140°C for 5 min. Utility of the thermotolerant yeast Kluyveromyces marxianus KCTC7150 was evaluated with respect to cell growth and ethanol fermentation at 40°C was close to optimal for enzymatic hydrolysis. This could lead to the integration of both the saccharification and fermentation processes. The levels of ethanol production by simultaneous saccharification and fermentation (SSF) with non-adapted and adapted K. marxianus KCTC7150 were 9.1 g/l with an ethanol yield (YEtOH) of 0.24 and 10.2 g/l with an ethanol yield (YEtOH) of 0.27 at 156 h, respectively. The two-phase SSF process was employed in this study to improve the efficiency of ethanol fermentation. Adapted K. marxianus KCTC7150 using the two-phase SSF process produced 13.5 g/l with an ethanol yield (YEtOH) of 0.35 at 96 h. Development of the two-phase SSF process could enhance the overall ethanol fermentation yields of the seaweed K. alvarezii.

Effect of Light with Different Wavelengths on Nostoc flagelliforme Cells in Liquid Culture

  • Dai, Yu-Jie;Li, Jing;Wei, Shu-Mei;Chen, Nan;Xiao, Yu-Peng;Tan, Zhi-Lei;Jia, Shi-Ru;Yuan, Nan-Nan;Tan, Ning;Song, Yi-Jie
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.534-538
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    • 2013
  • The effects of lights with different wavelengths on the growth and the yield of extracellular polysaccharides of Nostoc flagelliforme cells were investigated in a liquid cultivation. N. flagelliforme cells were cultured for 16 days in 500 ml conical flasks containing BG11 culture medium under $27{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ of light intensity and $25^{\circ}C$ on a rotary shaker (140 rpm). The chlorophyll a, phycocyanin, allophycocyanin, and phycoerythrin contents in N. flagelliforme cells under the lights of different wavelengths were also measured. It was found that the cell biomass and the yield of polysaccharide changed with different wavelengths of light. The biomass and the yield of extracellular polysaccharides under the red or violet light were higher than those under other light colors. Chlorophyll a, phycocyanin, and allophycocyanin are the main pigments in N. flagelliforme cells. The results showed that N. flagelliforme, like other cyanobacteria, has the ability of adjusting the contents and relative ratio of its pigments with the light quality. As a conclusion, N. flagelliforme cells favor red and violet lights and perform the complementary chromatic adaptation ability to acclimate to the changes of the light quality in the environment.

Production of Recombinant Human Hyperglycosylated Erythropoietin Using Cell Culture Technology by Improving Sialylation. (Sialic Acid 함량 증가 배양기술에 의한 재조합 인간 다당쇄 에리스로포이에틴의 생산)

  • 박세철;이승오;박만식;김승훈;김준환;송무영;이병규;고인영;강희일
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.142-148
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    • 2004
  • Erythropoietin is a main regulator of human erythropoiesis. Recombinant human erythropoietin (rhEPO) is one of the glycoproteins produced in animal cells, and it has oligo saccharides chains which comprise about 40% of its molecular mass. Because the content of sialic acid can extend circulatory lifetime, the high degree of sialylation is often a desirable feature of therapeutic glycoproteins. In this study, the sialylation of rhEPO produced by chinese hamster ovary cell culture was maximized by supplementing the culture medium with N-acetylm-annosamine (ManNAc), a direct intracellular precursor for sialic acid synthesis and 2-deoxy-2,3-dehydro-N-acetylneuraminic acid (NeuAc2en), a sialidase inhibitor. Feeding of 20 mM ManNAc/0.5 mM NeuAc2en into culture medium increased the sialic acid content by nearly tenfold compared with unsupplemented medium. This effect was achieved without affecting the cell growth or product yield. Six erythropoietin fractions differing in sialic acid content, ranging from 11∼15% of EPO, were identified from chinese hamster ovary cell-derived rhEPO by mono Q column chromatography. It was found that, at 20 mM ManNAc/0.5 mM NeuAc2en feeding, productivity of hyper-glycosylated EPO increased up to 50%, compared with the unsupplemented medium.

Utilization of cyclohexanol and characterization of Acinetobacter calcoaceticus C-15 (Acinetobacter calcoaceticus C-15에 의한 Cyclohexanol의 이용 및 그 특성)

  • Kim, Kyung Ae;Park, Jong Sung;Rhee, In Koo
    • Microbiology and Biotechnology Letters
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    • v.13 no.1
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    • pp.71-77
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    • 1985
  • A bacterium which grows on cyclohexanol as sole carbon and energy source was isolated from sludge of industrial areas in Taegu and identified as Acinetobacter calcoaceticus C-15. The growth medium for the optimal culture condition was composed of 0.2% cyclohexanol, 0.11% $NH_4Cl$, 0.05% $KH_2PO_4$, 0.2% $K_2HPO_4$, 0.02% $MgSO_4{\cdot}7H_2O$, and 0.05% yeast extracts. The optimal pH value and temperature for the growth were 7.2 and $33^{\circ}C$, respectively. Specific growth rate of A. calcoaceticus C-15 at $33^{\circ}C$ on the cyclohexanol and cyclohexanone was $0.27hr^{-1}$ and $0.15hr^{-1}$, respectively. Growth yield for cyclohexanol was 1.0. The bacteria utilized ethanol, 1-butanol, 1-pentanol, and cyclohexanol as a carbon source but not methanol, 1-hexanol, m-cresol, glycerol, and cyclohexane. The bacteria grew on benzoate, adipate, acetate, and citrate, but did not on salicylate, phthalate, p-hydroxybenzoate, and gluconate. A calcoaceticus C-15 did not utilize all kind of sugars other than xylose. Cell-free extracts contained $NAD^+$-linked cyclohexanol dehydrogenase which catalized the oxidation of cyclohexanol to cyclohexanone.

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Effects of Supplementing Brown Seaweed By-products in the Diet of Holstein Cows during Transition on Ruminal Fermentation, Growth Performance and Endocrine Responses

  • Hong, Z.S.;Kim, E.J.;Jin, Y.C.;Lee, J.S.;Choi, Y.J.;Lee, H.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.9
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    • pp.1296-1302
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    • 2015
  • This study was conducted to examine the effects of supplementing brown seaweed by-products (BSB) in the diet of ruminants on ruminal fermentation characteristics, growth performance, endocrine response, and milk production in Holstein cows. In Experiment 1, the effects of different levels (0%, 2%, and 4% of basal diet as Control, 2% BSB, 4% BSB, respectively) of BSB were evaluated at 3, 6, 9, 12, and 24 h in vitro batch culture rumen fermentation. The pH tended to be higher for the higher level of BSB supplementation, with the pH at 12 h being significantly higher (p<0.05) than that of the control. The concentration of ammonia nitrogen was lower at 3, 9, 12, and 24 h incubation (p<0.05) compared with the control, and tended to be low at other incubation times. Volatile fatty acid concentration appeared to be minimally changed while lower values were observed with 4% BSB treatment at 24 h (p<0.05). In Experiment 2, effects of levels (0%, 2%, and 4%) of BSB on growth performance, endocrine responses and milk production were studied with Holstein dairy cows during transition. Dry matter intake, daily gain and feed efficiency were not affected by BSB supplementation. The concentration of plasma estrogen for the control, 2% BSB and 4% BSB after three months of pregnancy were 55.7, 94.1, and 72.3 pg/mL, respectively (p = 0.08). Although the differences of progesterone levels between BSB treatments and the control were minimal, the concentration in 4% BSB treatment increased to 157.7% compared with the initial level of the study. Triiodothyronine and thyroxine levels were also higher after both three months and eight months of pregnancy than the initial level at the beginning of the study. In addition, BSB treatments during one month after delivery did not affect daily milk yield and composition. In conclusion, the present results indicate that supplementation of BSB did not compromise ruminal fermentation, and animal performance at lower levels and hence may have potential to be used as a safe feed ingredient in dairy cows.

Effects of NaCl on the Growth and Physiological Characteristics of Crepidiastrum sonchifolium (Maxim.) Pak & Kawano (NaCl 처리가 고들빼기의 생장과 생리적 특성에 미치는 영향)

  • Lee, Kyeong Cheol;Han, Sang Kyun;Yoon, Kyeong Kyu;Lee, Hak bong;Song, Jae Mo
    • Korean Journal of Medicinal Crop Science
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    • v.28 no.1
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    • pp.1-8
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    • 2020
  • Background: This study was conducted to investigate the effects of NaCl concentration on the photosynthetic parameters, chlorophyll fluorescence and growth characteristics of Crepidiastrum sonchifolium. Methods and Results: As treatments, we subjected C. sonchifolium plants to four different concentrations of NaCl (0, 50, 100 and 200 mM). We found that the photosynthetic parameters maximum photosynthesis rate (PN max), net apparent quantum yield (Φ), maximum carboxylation rate (Vcmax), and maximum electron transport rate (Jmax) were significantly reduced at an NaCl concentration greater than 100 mM. In contrast, there was an increase in water-use efficiency with increasing NaCl concentration, although in terms of growth performances, leaf dry weight, root dry weight, stem length, and total dry weight all decreased with increasing NaCl concentration. Furthermore, leakage of electrolytes, as a consequence of cell membrane damage, clearly increased in response to an increase in NaCl concentration. Analysis of the polyphasic elevation of chlorophyll a fluorescence transients (OKJIP) revealed marked decrease in flux ratios (ΦPO, ΨO and ΦEO) and the PIabs, performance index in response to treatment with 200 mM NaCl, thereby reflectings the relatively reduced state of photosystem II. This increase in fluorescence could be due to a reduction in electron transport beyond Q-A. We thus found that the photosynthetic parameters, chlorophyll fluorescence and growth characteristics of C. sonchifolium significantly increased in response to treatment with 200 mM NaCl. Conclusions: Collectively, the findings of this study indicate that C. sonchifolium shows relatively low sensitivity to NaCl stress, although photosynthetic activity was markedly reduced in plants exposed to 200 mM NaCl.

Ovarian Follicular Dynamics, Ovarian Follicular Growth, Oocyte Yield, In vitro Embryo Production and Repeated Oocyte Pick Up in Thai Native Heifers Undergoing Superstimulation

  • Chasombat, J.;Nagai, T.;Parnpai, R.;Vongpralub, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.4
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    • pp.488-500
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    • 2013
  • The objective of this study was to compare the effectiveness of the protocols for superstimulation of follicular growth in Thai native heifers. Heifers (n = 20) were randomly divided into four groups of five heifers/group. Heifers were given a single dose by i.m. administration of 100 mg Follicle Stimulating Hormone dissolved in polyvinylpyrrolidone (FSHp) at 24 h. Ovum pick up (OPU) occurred at 72 h ($F_{24}O_{72}$ protocol; Group 1) or 96 h ($F_{24}O_{96}$ protocol; Group 2), and at 36 h and OPU at 72 h ($F_{36}O_{72}$ protocol; Group 3) or 96 h ($F_{36}O_{96}$ protocol; Group 4) after follicular ablation. The dynamics of ovarian follicular growth were monitored by twice-daily ultrasonographic examinations. Blood sample collections were performed every 12 h after initiation of treatment for assessment of FSH, E2 and P4 profiles. All heifers were subjected to eight repeated sequential sessions of OPU. The follicular deviation commenced $24{\pm}5.32$ h after follicular ablation in all groups. The circulatory FSH surged quickly from 24 to 36 h (>0.8 ng/ml) after follicular ablation and circulatory estrogen levels steadily increased from 36 h until OPU in all groups. At the end of the OPU sessions, the mean number of aspirated follicles/heifer/session in $F_{36}O_{72}$ protocol (Group 3) and $F_{36}O_{96}$ protocol (Group 4) were higher than in the two other groups (p<0.05). The number of cumulus-oocyte complexes (COCs), cleaved and day 8 blastocysts rates in the $F_{36}O_{72}$ protocol (Group 3) were higher than in the other groups (p<0.05). It can be concluded that a single dose i.m. administration of 100 mg FSHp at 36 h and OPU at 72 h after follicular ablation ($F_{36}O_{72}$ protocol; Group 3) was the most effective protocol for superstimulation of follicular growth for repeated OPU and subsequent in vitro embryo production in Thai native heifers.

Environmental Effect on the Biodegradation of Toluene by Pseudomonas fluorescence KNU417 (원유오염 토양으로부터 분리한 Pseudomonas fluorescence KNU417의 톨루엔 분해에서 환경 인자의 영향)

  • Kwon, Hyeok-Man;Yeom, Sung-Ho
    • Journal of the Korea Organic Resources Recycling Association
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    • v.14 no.3
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    • pp.117-125
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    • 2006
  • A microorganism capable of degrading toluene was isolated from crude oil contaminated soil and identified as Pseudomonas fluorescence. The effects of environmental factors on the degradation of toluene were investigated. The optimum temperature for toluene degradation was $30^{\circ}C$ and the maximum specific cell growth and toluene degradation rates were $0.76hr^{-1}$ and $0.36hr^{-1}$, respectively. Although the wild cells were not able to degrade toluene at $10^{\circ}C$ and $40^{\circ}C$, the cells adapted to toluene at $30^{\circ}C$ degraded 100mg/L of toluene completely at $10^{\circ}C$ and 80% of the toluene at $40^{\circ}C$. The wild cells were not able to degrade more than 200mg/L of toluene but the toluene-adapted cells degraded up to 300mg/L of toluene. Although the optimum pH was 7.0, the degradation rates were not much different in the range of 5.5 to 9.0. When nitrate was used as a nitrogen source instead of ammonium, the adaptation period became longer by 2~10 hours and the cell growth yield became lower by 45%. The toluene degradation rates after adaptation period, however, were almost same in both cases. The observations in this study will be used in the future biofilter design and operation.

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Tylosin Production by Mutant Resistant to Oleic Acid (Oleic Acid 내성균주로부터 Tylosin 생산)

  • Choi, Du-Bok;Choi, On-You;Moon, Ok-Ran;Yoon, Mi-Ran;Ji, Sung-Nam;Shin, Dae-Yewn
    • Journal of Environmental Health Sciences
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    • v.31 no.5 s.86
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    • pp.360-364
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    • 2005
  • When rapeseed oil as the carbon source was used for tylosin production from Streptomyces fradiae TP-1239 was very sensitive to oleic acid. Cell growth was restrained by adding 0.8 g/l of oleic acid to the culture broth. Mutant strain TM-224-1 resistant to 1.2 g/l of oleic acid was obtained by screening in solid and liquid media containing oleic acid. The uptake rate of oleic acid by TM-224-1 was approximately 3.8 fold higher than the parent strain. For comparing the TM-224-1 and the parent strain, batch cultures were carried out in a jar fermentor. Cell growth of TM-224-1 strain was higher than the parent strain after two days of culturing. However, after four days of culturing, it was similar to that of the parent strain. The amount of rapeseed oil consumed by TM-224-1 and the parent strain were 60.5 and 78.2 g/l, respectively. The production and yield of tylosin was aproximately 2.0 and 3.2 fold higher than the parent strain, respectively. From these results, it was concluded that this mutant, which was resistant to oleic acid, has improved tylosin production.