• Title/Summary/Keyword: AS/ES

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Suspension Culture-Mediated Tetraploid Formation in Mouse Embryonic Stem Cells

  • Lee, Jae-Hee;Gong, Seung-Pyo;Lim, Jeong-Mook;Lee, Seung-Tae
    • Reproductive and Developmental Biology
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    • v.36 no.1
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    • pp.21-26
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    • 2012
  • Suspension culture is a useful tool for culturing embryonic stem (ES) cells in large-scale, but the stability of pluripotency and karyotype has to be maintained $in$ $vitro$ for clinical application. Therefore, we investigated whether the chromosomal abnormality of ES cells was induced in suspension culture or not. The ES cells were cultured in suspension as a form of aggregate with or without mouse embryonic fibroblasts (MEFs), and 0 or 1,000 U/ml leukemia inhibitory factor (LIF) was treated to suspended ES cells. After culturing ES cells in suspension, their karyotype, DNA content, and properties of pluripotency and differentiation were evaluated. As a result, the formation of tetraploid ES cell population was significantly increased in suspension culture in which ES cells were co-cultured with both MEFs and LIF. Tetraploid ES cell population was also generated when ES cells were cultured alone in suspension regardless of the existence of LIF. On the other hand, the formation of tetraploid ES cell population was not detected in LIF-free condition, in which MEFs were included. The origin of tetraploid ES cell population was turned out to be E14 ES cells and not MEFs by microsatellite analysis and the basic properties of them were still maintained despite ploidy-conversion to tetraploidy. Furthermore, we identified the ploidy shift from tetraploidy to near-triploidy as tetraploid ES cells were differentiated spontaneously. From these results, we demonstrated that suspension culture system could induce ploidy-conversion generating tetraploid ES cell population. Moreover, optimization of suspension culture system may make possible mass-production of ES cells.

The effect of eleutherococcus senticosus on metabolism-associated protein expression in 3T3-L1 and C2C12 cells

  • Hashimoto, Takeshi;Okada, Yoko;Yamanaka, Atsushi;Ono, Natsuhiko;Uryu, Keisuke;Maru, Isafumi
    • Korean Journal of Exercise Nutrition
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    • v.24 no.3
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    • pp.13-18
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    • 2020
  • [Purpose] In vivo studies have demonstrated the ergogenic benefits of eleutherococcus senticosus (ES) supplementation. ES has been observed to enhance endurance capacity, improve cardiovascular function, and alter metabolic functions (e.g., increased fat utilization); however, the exact mechanisms involved remain unknown. We aimed to determine whether ES could effectively induce fat loss and improve muscle metabolic profiles through increases in lipolysis- and lipid metabolism-associated protein expression in 3T3-L1 adipocytes and C2C12 skeletal muscle cells, respectively, to uncover the direct effects of ES on adipocytes and skeletal muscle cells. [Methods] Different doses of ES extracts (0.2, 0.5, and 1.0 mg/mL) were added to cells (0.2 ES, 0.5 ES, and 1.0 ES, respectively) for 72 h and compared to the vehicle control (control). [Results] The intracellular triacylglycerol (TG) content significantly decreased (p < 0.05 for 0.2 ES, p < 0.01 for 0.5 ES and 1.0 ES) in 3T3-L1 cells. Adipose triglyceride lipase, which is involved in active lipolysis, was significantly higher in the 1.0 ES group than in the control group (p < 0.01) of 3T3-L1 adipocytes. In C2C12 cells, the mitochondrial protein voltage-dependent anion channel (VDAC) was significantly increased in the 1.0 ES group (p < 0.01). Furthermore, we found that 1.0 ES activated both 5' AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) in skeletal muscle cells (p < 0.01). [Conclusion] These findings suggest that ES extracts decreased TG content, presumably by increasing lipase in adipocytes and metabolism-associated protein expression as well as mitochondrial biogenesis in muscle cells. These effects may corroborate previous in vivo findings regarding the ergogenic effects of ES supplementation.

Value Chain Network of Environment and Safety for the Industry : Its Necessity and Disclosure (산업환경·생산안전 가치창출 네트워크 구축 필요성과 담론)

  • Kim, Jae Youn;Lee, Hankyung
    • Clean Technology
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    • v.22 no.1
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    • pp.62-72
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    • 2016
  • These days, it seems to be a core time to build proactive prevention systems for small and medium sized enterprises due to the reinforced safety in the production process and the strong environmental regulations (ES) both at home and abroad. On the other hand, a network of experts that combines the industrial environment and production safety as well as even the knowledge services companies are not quite enough to prepare to support them. In this study, through a survey of experts the current statue of the convergence of industrial environment and production safety were reviewed, and the structure of knowledge ES cluster was proposed to overcome this current state. Detailed strategies such as the development and distribution of ES convergence methodology, ES efficiency analysis and an ES roundtable.

Follow Up Expression Patterns of Alkaline Phosphatase(AP) as a Marker for Establishing Mouse Embryonic Stem (ES) Cells (배아주간세포수립을 위한 Alkaline Phosphatase(AP)의 상이한 발현 양식의 추적)

  • 김진회;차수경;노민경;송상진;구덕본;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.19 no.1
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    • pp.55-63
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    • 1995
  • The putative totipotency germ cells has a relative abundance of alkaline phosphatases. Thus, histological staining of AP activity offers a new route to isolate totipotent cells and also provides insights into culture systems of these cells. Furthermore, the AP staining technique is simple and fast, requires only the napthol AS/MS substrate in combination with trapping diazonium salts such as fast red or fast blue. However, our unexpected finding was that AP staining of mouse ES cells were detected in the undifferentiaed epiblast-derived cells as well as several types of differentiating cells. This findings are different from results of Talbot et al. (1993) reported usefulness of the AP staining and implies that histological staining of AP may not by useful to determine undifferentiaed state or totipotency of ES cells. Thus, we have investigated the patterns of AP expression by RT-PCR in order to identify a marker of undifferentiated ES/primordial germ (PG) cells. In RT-PCR analysis, embryonic (E)-AP was detected only in undifferentiated ES cells, but intestinal(I)-AP was not detected in all of the examined ES and PG cells. In addition, nonspecific (NS)-AP wasdetected in undifferentiated PG cell from day 7, 5 to 13 of gestation. Histological activity of AP in ES cells was completely suppressed by addition of L-phenylalanine (Phe), L-homoarginine (Har), and L-phenylalanylglycylglycine (PheGlyGly) as an inhibitor, but RT-PCR showed the same results as in the absence of an inhibitors. Our findings suggested that expression of E-AP and NS-AP may use as a marker to determine the undifferentiated status in ES and PG cells.

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Electrospray technique for preparation of core-shell materials : A mini-review

  • Tran, Vinh Van;Lee, Young-Chul
    • Particle and aerosol research
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    • v.14 no.3
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    • pp.49-63
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    • 2018
  • During the last decade, electrospray (ES) techniques have been used as potential methods for preparing of core-shell materials. Depending on the architecture of nozzle and design of devices, the ES techniques includes monoaxial, coaxial, multiple coaxial nozzle ES and microfluidic ES devices. ES operates based on a basic principle, in which a spray of monodisperse droplets is formed by dispensing an electrically conductive liquid through a capillary charged to a sufficiently high potential. In review of many recent research papers, we take a closer look at ES techniques and their applications for fabrication of core-shell materials. Several advantages of ES technique compared with other methods were emphasized and it may be regarded as a potential tool for fabrication of core-shell materials current and near future.

Induction of Myogenic Differentiation in Myoblasts by Electrical Stimulation

  • Je, Hyeon-Jeong;Kim, Min-Gu;Cho, Il-Hoon;Kwon, Hyuck-Joon
    • Journal of the Korean Society of Physical Medicine
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    • v.14 no.2
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    • pp.63-70
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    • 2019
  • PURPOSE: While electrical stimulation (ES) is known to be a safe and flexible tool in rehabilitation therapy, it has had limited adoption in muscle regeneration. This study was performed to investigate whether ES can induce myogenic differentiation and to clarify the mechanism underlying the effects of ES on myogenic differentiation. METHODS: This study used rat L6 cell lines as myoblasts for myogenic differentiation. Electric stimulation was applied to the cells using a C-Pace EP culture pacer (IonOptix, Westwood, Ma, USA). The gene expressions of myogenic markers were examined using qPCR and immunochemistry. RESULTS: Our study showed that ES increased the thickness and length of myotubes during myogenic differentiation. It was found that ES increased the expression of myogenic markers, such as MyoD and Myogenin, and also activated the fusion of the myoblast cells. In addition, ES suppressed the expression of small GTPases, which can explain why ES promotes myogenic differentiation. CONCLUSION: We found that ES induced myogenic differentiation by suppressing small GTPases, inhibiting cell division. We suggest that ES-based therapies can contribute to the development of safe and efficient muscle regeneration.

Differentiated Human Embryonic Stem Cells Enhance the In vitro and In vivo Developmental Potential of Mouse Preimplantation Embryos

  • Kim, Eun-Young;Lee, Keum-Sil;Park, Se-Pill
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.9
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    • pp.1152-1158
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    • 2010
  • In differentiating human embryonic stem (d-hES) cells there are a number of types of cells which may secrete various nutrients and helpful materials for pre-implantation embryonic development. This study examined whether the d-hES could function as a feeder cell in vitro to support mouse embryonic development. By RT-PCR analysis, the d-hES cells revealed high expression of three germ-layered differentiation markers while having markedly reduced expression of stem cell markers. Also, in d-hES cells, LIF expression in embryo implantation-related material was confirmed at a similar level to undifferentiated ES cells. When mouse 2PN embryos were cultured in control M16 medium, co-culture control CR1aa medium or co-cultured with d-hES cells, their blastocyst development rate at embryonic day 4 (83.9%) were significantly better in the d-hES cell group than in the CR1aa group (66.0%), while not better than in the M16 group (90.7%)(p<0.05). However, at embryonic days 5 and 6, embryo hatching and hatched-out rates of the dhES cell group (53.6 and 48.2%, respectively) were superior to those of the M16 group (40.7 and 40.7%, respectively). At embryonic day 4, blastocysts of the d-hES cell group were transferred into pseudo-pregnant recipients, and pregnancy rate (75.0%) was very high compared to the other groups (M16, 57.1%; CR1aa, 37.5%). In addition, embryo implantation (55.9%) and live fetus rate (38.2%) of the d-hES cell group were also better than those of the other groups (M16, 36.7 and 18.3%, respectively; CR1aa, 23.2 and 8.7%, respectively). These results demonstrated that d-hES cells can be used as a feeder cell for enhancing in vitro and in vivo developmental potential of mouse pre-implantation embryos.

Effects of Using Soybean Protein Emulsion as a Meat Substitute for Chicken Breast on Physicochemical Properties of Vienna Sausage

  • Kang, Kyu-Min;Lee, Sol-Hee;Kim, Hack-Youn
    • Food Science of Animal Resources
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    • v.42 no.1
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    • pp.73-83
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    • 2022
  • The aim of this study is to determine the effects of using emulsion manufactured with soybeans (ES) to substitute chicken breast in Vienna sausages. Four types of Vienna sausages (S1: 10% ES and 50% chicken, S2: 20% ES and 40% chicken, S3: 30% ES and 30% chicken, and S4: 40% ES and 20% chicken) for this study were made. The pH, color, proximate composition, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), microphotographs, cooking yields, and texture profile analysis of sausages were examined. The pH value of uncooked and cooked sausages increased significantly with increasing ES content (p<0.05). The crude protein contents of S2, S3, and S4 were significantly higher than that of the control (p<0.05). Furthermore, the SDS-PAGE results showed that α-conglycinin, β-conglycinin, and the acidic subunit of glycinin all increased with increasing ES content. Microphotographs revealed that increasing the ES content decreased the size of fat globules. The cooking yields of samples increased significantly with increasing ES content (p<0.05). The hardness values of ES treated samples were significantly lower than that of the control (p<0.05). Therefore, 30% substitute of chicken breast with ES can improve the quality and structure of Vienna sausage, without inducing critical defects.

Electrical Properties of Polyaniline according to Preparation Conditions (제조 조건에 따른 Polyaniline의 전기적 성질)

  • 김언령;김태영;이보현;김종은;서광석;배종현
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.14 no.3
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    • pp.215-222
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    • 2001
  • Polyaniline-Camphorsulfonic Acid Emeraldine Salt(PANI-CSA ES) was prepared by doping Polyaniline Ermeralidine Base(PANI EB) with DL-10-Camphorsulfonic Acid(CSA). PANI-CSA ES was solved in an organic solvent by ultrasonification for different periods of time and its surface resistivity was measured. Several PANI-CSA ES solutions solved in different organic solvents were prepared and their surface resistivities were measured. Thermal stability of film casted with PANI-CAS ES solution in m-cresol was estimated by measuring its surface conductivity and the content of this moisture and organic solvents. PANI-CSA ES was blended with different polymeric binders to improve its physical properties and the surface resistivities of several kinds of PANI-CSA ES blends were measured as a function of the content of PANI-CSA ES. PANI-CSA ES polymerized by 1-step oxidative polymerization was prepared and its surface resistivity was measured.

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Controlling the Gene Expression of Corynebacterium diphtheria Toxin-A Using the Tet-On System in Mouse Embryonic Stem Cells. (Mouse Embryonic Stem Cell에서 Tetracycline-Inducible System(Tet-on System)을 이용한 Corynebacterium diphtheria Toxin-A유전자의 발현 조절)

  • 박재균;임수빈;송지환
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.11-15
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    • 2004
  • Embryonic stem (ES) cells are derived from the inner cell mass of the blastocyst-stage embryos that can be propagated indefinitely and, at the same time, can be differentiated into all the cell types that constitute the body. Current research using ES cells is mainly focused on the efficient generation of specific cell types by employing optimal differentiation conditions, which often requires the genetic manipulation of ES cells. As a way of developing an efficient system to regulate foreign gene expression in ES cells, we have inserted the gene encoding Corynebacterium diphtheria toxin-A (DTA) into an autonomously induced plasmid under positive doxycycline control ('Tet-on' system). In this study, we demonstrate that this system can lead to the cell death of mouse ES cells by the induction of DTA expression when exposed to the tetracycline derivative, doxycycline. MTT assay showed that this induction resulted in the apoptosis of ES cells.