• Title/Summary/Keyword: cell production

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Production of Identical Multiplets from Deep-Frozen Embryos (동결보존배를 이용한 일란성 다태생산에 관한 연구)

  • Shin Sang-Tae
    • Journal of Veterinary Clinics
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    • v.10 no.2
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    • pp.243-249
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    • 1993
  • Although plenty of experiments for production of genetically identical twins have been reported, most of these reports were focused on microsurgical bisection of morula stage embryos, and little research has been performed for the production with frozen emb교os. The objective of these experiments were to assess the in vitro and in vitro developmental potential of blastomeres isolated from frozen-thawed 4-cell and 8-cell mouse embryos and to produce the identical multiplets from those embryos. The percentages of isolated 1/4, 2/4, 4/4, (control), 1/8, 2/8, 3/8, 4/8 and 8/8(control) blastomeres of 4-cell and 8-cell mouse emb교os which developed to normal blastocyst were 38.7%, 73.6%, 96.2(control), 15.1%, 40.1%, 65.9%, 88.3%, and 98.5%(control), respectively. The percentages of isolated 1/4, 2/4, 4/4(control), 1/8, 2/8, 3/8, 4/8 and 8/8(control) blastomeres of frozen-thawed 4-cell and 8-cell mouse embryos which developed to normal blastocyst were 35.6%, 68.5%, 100.0%(control), 16.1%, 50.6%, 71.7%, 90.9% and 100.0%(control), respectively. Normal 26 pups were obtained by transfer of 240 blastocyts developed 1, on 1/4 to 8/8 blastomeres. Those 26 pups obtained, 4 identical twins were produced from 2/4, 2/8, 3/8 and 4/8 blastomeres.

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THE EFFECTS OF MECHANICAL STRESS ON CULTURED BONE CELL POPULATIONS (Mechanical stress가 골조직세포군에 미치는 영향)

  • Kim, Sang-Tae;Cha, Kyung-Suk
    • The korean journal of orthodontics
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    • v.24 no.1 s.44
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    • pp.105-114
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    • 1994
  • The movement of teeth during orthodontic treatment requires bone remodeling process of bone formation and bone resolution. To find out the changes occuring in the cell itself, mechanical stress was applied to the cell populations involved in the bone metabolism. Bone tissue cell populations were isolated from fetal rat calvaria and divided into OC and OB groups. Following results were obtained from measuring the changes in acid & alkaline phosphatease activity, cyclic AMP and $PGE_2$ production in time lapse after the application of mechanical stress. 1. In case of the marker enzyme of specific bone tissue cell, acid phosphatase activity was high in OC group and alkaline phosphatase activity was high in OB group. 2. After the mechanical stress was applied, acid phosphatase activity was decreased in both OC and OB groups and alkaline phosphatase activity was increase in OB group. 3. When the mechanical stress was applied for 15, 30 and 60 minutes, the production of $PGE_2$ increased in both OC and OB groups, as the time span increased. 4. When the mechanical stress was applied for 20 and 40 minutes, the production of $PGE_2$ increased in both OC and OB groups, as the time span increased.

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Production of Novel Cell-Associated Tannase from Newly Isolated Serratia ficaria DTC

  • Belur, Prasanna D.;Gopal, Mugeraya;Nirmala, K.R.;Basavaraj, N.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.732-736
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    • 2010
  • Five strains of tannic acid degrading bacteria were isolated and identified by phenotypic characterization. All the five isolates showed cell-associated activity, whereas only three showed extracellular activity. Serratia ficaria DTC, showing the highest cell-associated activity (0.29 U/l), was selected for further shake-flask studies. Tannase synthesis was growth associated and reached the peak in the late stationary phase of growth. Organic nitrogen sources enhanced the tannase production. Peak tannase production of 0.56 U/l was recorded in the medium having the initial pH of 6. The pH and temperature optima of the enzyme were found to be 8.9 and $35^{\circ}C$, respectively. This is the first report of cell-associated activity in the case of bacterial tannase. Cell-associated tannase of Serratia ficaria DTC could be industrially important from the perspective of its activity at broad temperature and pH ranges, and its unusually high activity at pH 8.9.

Whole-Cell Biocatalysis for Producing Ginsenoside Rd from Rb1 Using Lactobacillus rhamnosus GG

  • Ku, Seockmo;You, Hyun Ju;Park, Myeong Soo;Ji, Geun Eog
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1206-1215
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    • 2016
  • Ginsenosides are the major active ingredients in ginseng used for human therapeutic plant medicines. One of the most well-known probiotic bacteria among the various strains on the functional food market is Lactobacillus rhamnosus GG. Biocatalytic methods using probiotic enzymes for producing deglycosylated ginsenosides such as Rd have a growing significance in the functional food industry. The addition of 2% cellobiose (w/v) to glucose-free de Man-Rogosa-Sharpe broths notably induced β-glucosidase production from L. rhamnosus GG. Enzyme production and activity were optimized at a pH, temperature, and cellobiose concentration of 6.0, 40℃, and 2% (w/v), respectively. Under these controlled conditions, β-glucosidase production in L. rhamnosus GG was enhanced by 25-fold. Additionally, whole-cell homogenates showed the highest β-glucosidase activity when compared with disrupted cell suspensions; the cell disruption step significantly decreased the β-glucosidase activity. Based on the optimized enzyme conditions, whole-cell L. rhamnosus GG was successfully used to convert ginsenoside Rb1 into Rd.

The Effects of Dietary Enzyme Mixture Reinforced with ${\beta}$-Glucanase Activity on Mini Production and the Change of Somatic Cell Count in Lactating Dairy Cows (사료내 ${\beta}$-glucanase 활성 강화 고역가 복합효소제 첨가급여가 착유우의 유생산 및 체세포수 변화에 미치는 영향)

  • Joo, Eun-Jung;Jeong, Su-Jin;Yoon, Byung-Seon;Nam, Ki-Taek;Choi, Il-Shin;Ahn, Jong-Ho;Hwang, Seong-Gu
    • Korean Journal of Organic Agriculture
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    • v.12 no.2
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    • pp.231-241
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    • 2004
  • In recent years, many researches are actively undertaken for environmental-friendly animal production according to the increased understanding about food safety because of the outbreak of various diseases such as mad cow disease, Foot and mouth disease and Poultry Influenza virus. However, high quality(higher safety)- animal production may not be successful without increasing of disease resistance of animal and the improvement of feeding environment. To increase the disease resistance is able to be accomplished by stimulating the immune function. The present study was undertaken to investigate the effects of enzyme mixture reinforced with ${\beta}$-glucanase activity which degrade polysaccharide to release ${\beta}$-glucan known as stimulator of immune function on the change of milk production and somatic cell count. After 12weeks of experimental feeding, milk production tended to be increased and somatic cell count was decreased from average $227{\times}10^4$ to $37.1{\times}10^4$. Milk protein and solid-fat content were tended to increase but milk fat showed decreasing tendency by the feeding of enzyme mixture. All together, it has been suggest6d that the improvement of high quality milk production may be possible through the dietary addition of immune modulating enzyme mixture in lactating dairy cows.

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Effects of Galgeungyulpitang on Cellular Production of Melanin and Elastase

  • Jo, Na Young;Lee, Eun Yong;Lee, Cham Kyul;Roh, Jeong Du
    • Journal of Acupuncture Research
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    • v.36 no.1
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    • pp.33-37
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    • 2019
  • Background: This study was designed to investigate the potential effects of Galgeungyulpitang for whitening and elasticity treatment by examining its effect on melanoma cells. Methods: The effects of Galgeungyulpitang on B16/F10 melanoma cell viability, production of melanin, tyrosinase and elastase, were investigated. Cell viability was measured by colorimetric assay that assesses cell metabolic activity (MTT assay). Melanin was measured by Hosei's method, tyrosinase was measured by Yogi's method and elastase was measured by James's method. Results: At concentrations higher than $500{\mu}g/mL$ Galgeungyulpitang, cell viability was significantly reduced ($p{\leq}0.05$). At concentrations of $500{\mu}g/mL$ and lower, morphological changes were not observed. The rate of melanin synthesis was significantly reduced to $73.49%{\pm}2.92%$ at a concentration of $500{\mu}g/mL$ Galgeungyulpitang compared with untreated cells (p < 0.05). Extracellular tyrosinase production was not significantly decreased in vitro, however, intracellular tyrosinase production was significantly reduced to $76.06%{\pm}2.17%$ when treated with Galgeungyulpitang at a concentration of $500{\mu}g/mL$ compared with the control (p < 0.05). Elastase Type 1 production was significantly reduced to $74.98%{\pm}3.24%$ and $69.62%{\pm}4.66%$ at concentrations of 250 and $500{\mu}g/mL$ Galgeungyulpitang, respectively (p < 0.05). Elastase Type 4 production was significantly reduced to $72.77%{\pm}3.52%$ at concentrations of 250 and $500{\mu}g/mL$ (p < 0.05). Conclusion: The results in this study showed that Galgeungyulpitang may inhibit melanin and tyrosinase synthesis, and inhibit elastase production, suggesting that Galgeungyulpitang may be potentially beneficial for skin whitening and loss of skin elasticity treatments.

A Novel Oxidative Stress-inducible Peroxidase Promoter and Its Applications to Production of Pharmaceutical Proteins in Transgenic Cell Cultures

  • Lee, Ok-Sun;Park, Sun-Mi;Kwon, Suk-Yoon;Lee, Haeng-Soon;Kim, Kee-Yeun;Kim, Jae-Whune;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.143-150
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    • 2002
  • A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

Production of 10-deacetylbaccatin III in Taxus cuspidata Suspension Cell Culture (주목 현탁세포배양을 이용한 10-deacetylbaccatin III 생산)

  • Lee, Gue-Wha;Kim, Dong-Il
    • KSBB Journal
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    • v.14 no.6
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    • pp.665-671
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    • 1999
  • In this study, enhanced production of 10-deacetylbaccatin III(10-DAB), a precursor of taxol in semisynthesis, was investigated in Taxus cuspidata cell suspension cultures. The effects of initial inoculum size and sugar concentration were examined to prove the relationship between the production of 10-DAB and cell growth. The cell growth was found to be stimulated in Schenk and Hildebrandt(SH) medium. The lower the inoculum size as well as initial sugar concentration, the faster the cell growth rate. When the initial sugar concentration was dept low, the production of 10-DAB into medium was increased. By using perfusion culture, continuous cell growth was possible until the end of culture and more than 34.67 g/L of cell concentration could by obtained. This is about 2.5 times higher level than that of control batch culture.

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Rumen Microbes, Enzymes and Feed Digestion-A Review

  • Wang, Y.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.11
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    • pp.1659-1676
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    • 2002
  • Ruminant animals develop a diverse and sophisticated microbial ecosystem for digesting fibrous feedstuffs. Plant cell walls are complex and their structures are not fully understood, but it is generally believed that the chemical properties of some plant cell wall compounds and the cross-linked three-dimensional matrix of polysaccharides, lignin and phenolic compounds limit digestion of cell wall polysaccharides by ruminal microbes. Three adaptive strategies have been identified in the ruminal ecosystem for degrading plant cell walls: production of the full slate of enzymes required to cleave the numerous bonds within cell walls; attachment and colonization of feed particles; and synergetic interactions among ruminal species. Nonetheless, digestion of fibrous feeds remains incomplete, and numerous research attempts have been made to increase this extent of digestion. Exogenous fibrolytic enzymes (EFE) have been used successfully in monogastric animal production for some time. The possibility of adapting EFE as feed additives for ruminants is under intensive study. To date, animal responses to EFE supplements have varied greatly due to differences in enzyme source, application method, and types of diets and livestock. Currently available information suggests delivery of EFE by applying them to feed offers the best chance to increase ruminal digestion. The general tendency of EFE to increase rate, but not extent, of fibre digestion indicates that the products currently on the market for ruminants may not be introducing novel enzyme activities into the rumen. Recent research suggests that cleavage of esterified linkages (e.g., acetylesterase, ferulic acid esterase) within the plant cell wall matrix may be the key to increasing the extent of cell wall digestion in the rumen. Thus, a crucial ingredient in an effective enzyme additive for ruminants may be an as yet undetermined esterase that may not be included, quantified or listed in the majority of available enzyme preparations. Identifying these pivotal enzyme(s) and using biotechnology to enhance their production is necessary for long term improvements in feed digestion using EFE. Pretreating fibrous feeds with alkali in addition to EFE also shows promise for improving the efficacy of enzyme supplements.

Apoptotic Effect of ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts on MIA PaCa-2 Cells (인진과 황련 추출물의 췌장암 세포주 MIA PaCa-2에 대한 세포사멸 효과)

  • Joo, Hyun-A;Bae, Hyeon-Jin;Hwang, Chung-Yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.27 no.4
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    • pp.158-176
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    • 2014
  • Purpose : The purpose of this study is to investigate the effect of ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts on cell death in pancreatic cancer cells. Method : Human-derived pancreatic cancer cell line, MIA PaCa-2 cells were treated by Prescription A with various concentrations and the cytotoxicity was determined by MTT assay. To investigate the effects of Prescription A on pancreatic cancer cells, the staining of Annexin V/PI, cell cycle arrest, nuclear chromatin condensation and the production of reactive oxygen species (ROS) were examined. The effect of Prescription A's effective components, ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts on cell death were also observed. Results : The viability of MIA PaCa-2 cells treated with Prescription A were decreased in a dose dependent manner. Prescription A induced cell death in MIA PaCa-2 cells as shown by result of Annexin V/PI double staining, chromatin condensation and cell cycle arrest. In addition, production of ROS was increased by Prescription A treatment, suggesting that ROS induced by Prescription A mediated cell death. Furthermore, Prescription A's effective components, ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts were also induced apoptosis of MIA PaCa-2 cells through ROS production. Conclusion : These results suggest that Prescription A's effective components, ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts induced death of MIA PaCa-2 through ROS production.