• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.1-8
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• 2011

Techniques to evaluate gene expression profiling, such as sufficiently sensitive cDNA microarrays or real-time quantitative PCR, are efficient methods for monitoring human pluripotent stem cell (hESC/iPSC) cultures. However, most of these high-throughput tests have a limited use due to high cost, extended turn-around time, and the involvement of highly specialized technical expertise. Hence, there is an urgency of rapid, cost-effective, robust, yet sensitive method development for routine screening of hESCs/hiPSCs. A critical requirement in hESC/hiPSC cultures is to maintain a uniform undifferentiated state and to determine their differentiation capacity by showing the expression of gene markers representing all three germ layers, including ectoderm, mesoderm, and endoderm. To quantify the modulation of gene expression in hESCs/hiPSC during their propagation, expansion, and differentiation via embryoid body (EB) formation, we developed a simple, rapid, inexpensive, and definitive multimarker, semiquantitative multiplex RT-PCR platform technology. Among the 9 gene primers tested, 5 were pluripotent markers comprising set 1, and 3 lineage-specific markers were combined as set 2, respectively. We found that these 2 sets were not only effective in determining the relative differentiation in hESCs/hiPSCs, but were easily reproducible. In this study, we used the hES/hiPS cell lines to standardize the technique. This multiplex RT-PCR assay is flexible and, by selecting appropriate reporter genes, can be designed for characterization of different hESC/hiPSC lines during routine maintenance and directed differentiation.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.33-33
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• 2003

This study was conduct to compare the efficacy to produce male and female somatic cloned piglets. Maturation of porcine COCs was accomplished by incubation in NCSU-23 medium supplemented with 0.6 mM cysteine, 10% porcine follicular fluid, 1mM dibutyryl cyclic adenosine monophosphate (dbc-AMP, Sigma, USA), and 0.1 IU/ml human menopausal gonadotrophin (hMG, Teikokuzoki, Japan) for 20h and then cultured without dbcAMP and hMG for another 18 to 24 h. Female and male fetal cells were isolated from each fetus, cultured in ES-DMEM medium containing 10% FCS. Enucleated oocytes were fused with fetal fibroblasts (passage 4 to 15). Reconstructed embryos were cultured in NCSU-23 with 4 mg/ml BSA under mineral oil at 39$^{\circ}C$ in 5% $CO_2$ in air. A total of 12,328 nuclear-transferred embryos (1- to 4-cell stage) were surgically transferred into 69 surrogate gilts. Three recipients aborted during the period of conception. Three gilts delivered eleven female piglets, and five recipients gave rise to birth 22 male piglets. The average birth weigh of the cloned piglets was 1.52 kg (1.38~1.83 kg) in female piglets and 0.84 kg (0.45~1.25 kg) in male piglets. Alive cloned pigs was seven in female piglets (63.6%) and four in male piglets (18.2%). The other two recipients is ongoing. This study suggests that female-derived fetal cell as a nuclear donor has more capability on production of cloned piglets than male.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.75-75
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• 2003

Since the establishment of embryonic stem cell, pluripotency of the cells was known to allow differentiation of the cells into various cell types consisting whole body. Several protocols have been developed to induce expression of specific genes.. However, no precise protocol that will generate a single type of the cells from stem cells has been reported. In order to produce cells suitable for transplantion into brain of PD animal model, which arouse due to a progressive degeneration of dopaminergic neurons in midbrain, human embryonic stem cell (hESC, MB03) was transfected with cDNAs cording for tyrosine hydroxylase (TH). Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by the two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA/ascorbic acid (AA), embryoid bodies (EB, for 4days) derived from hES cells were exposed to RA (10$^{-6}$ M)/AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. By indirect immunocytochemical studies, proportion of cells expressing NF200 increased rapidly from 20% at 7 days to 70 % at 28 days in RA/AA-treated group, while those cells expressing NF160 decreased from 80% at 7 days to 10% at 28 days upon differentiation in N2 medium. However, in differentiation by RA/AA treatment system, there was a significant increase in proportion of neuron maturity (73%) at day 14 after N2 medium. TH#2/MB03 cells expressing TH are >90% when matured at the absence of either bDNF or TGF-$\alpha$. These results suggested that TH#2/MB03 cells could be differentiated in vitro into mature neurons by RA/AA.

• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.739-747
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• 2018

The low solubility and high-cost recovery of Propionibacterium acnes polyunsaturated fatty acid isomerase (PAI) are key problems in the bioproduction of high value-added conjugated linoleic acid (CLA). To improve the solubility of recombinant PAI, six chaperone proteins were coexpressed with PAI. Introduction of GroELS proteins dramatically improved the PAI solubility from 29% to 97%, with increased activity by 57.8%. Combined expression of DnaKJ-GrpE and GroELS proteins increased the activity by 11.9%. In contrast, coexpression of DnaKJ-GrpE proteins significantly reduced the activity by 57.4%. Plasmids pTf16 harboring the tig gene and pG-Tf2 containing the tig and groEL-groES genes had no visible impact on PAI expression. The lytic protein E was then introduced into the recombinant Escherichia coli to develop a cell autolysis system. A 35% activity of total intracellular PAI was released from the cytoplasm by suspending the lysed cells in distilled water. The PAI recovery was further improved to 81% by optimizing the release conditions. The lipase from Rhizopus oryzae was also expressed in E. coli, with an extracellular activity of 110.9 U/ml. By using the free PAI and lipase as catalysts, a joint system was established for producing CLA from sunflower oil. Under the optimized conditions, the maximum titer of t-10,c-12-CLA reached 9.4 g/l. This work provides an effective and low-cost strategy to improve the solubility and recovery of the recombinant intracellular PAI for further large-scale production of CLA.

• Bulletin of the Korean Chemical Society
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• v.28 no.12
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• pp.2261-2265
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• 2007

Prevention of thermal aggregation of the denatured protein by the group II chaperonin from the aerobic hyperthermophilic crenarchaeon Aeropyrum pernix K1 (ApcpnA) has been investigated. ApcpnA exists as a homo-oligomer in a ring structure, which protects thermal aggregation of the chemically denatured bovine rhodanese at 50 oC. ApcpnA alone is not sufficient for chaperonin activity, but the chaperonin activity is greatly enhanced in the presence of manganese ion and ATP. Compared to the mesophilic chaperonin GroEL/GroES, ApcpnA is more activated at a higher temperature and protects the aggregation-prone unfolded state of the denatured rhodanese from thermal aggregation. Binding of ATP is sufficient for ApcpnA to perform the chaperonin function in vitro, but hydrolysis of ATP is not necessarily required. We propose that utilization of Mn2+ and adenosine nucleotide regardless of ATP hydrolysis may be one of peculiar properties of archaeal chaperonins.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2011.05a
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• pp.140-142
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• 2011

In this paper, special purpose arithmetic unit for mobile graphics accelerator is designed. The designed processor supports six operations, such as $1/{\chi}$, $\frac{1}{{\sqrt{x}}$, $log_2x$, $2^x$, $sin(x)$, $cos(x)$. The processor adopts 2nd-order polynomial minimax approximation scheme based on IEEE floating point data format to satisfy accuracy conditions and has 5-stage pipeline structure to meet high operational rates. The SFAU processor consists of 23,000 gates and its estimated operating frequency is about 400 Mhz at operating condition of 65nm CMOS technology. Because the processor can execute all operations with 5-stage pipeline scheme, it has about 400 MOPS(million operations per second) execution rate. Thus, it can be applicable to the 3D mobile graphics processors.

• Journal of the Korea Institute of Information and Communication Engineering
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• v.13 no.4
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• pp.727-736
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• 2009

In this paper, the $arccos(cos^{-1})$ arithmetic unit for mobile graphics accelerator is designed. The mobile vector graphics applications need tight area, execution time, power dissipation, and accuracy constraints compared to desktop PC applications. The designed processor adopts 2nd-order polynomial approximation scheme based on IEEE floating point data format to satisfy speed and accuracy conditions and reduces area via hardware sharing structure. The arccosine processor consists of 15,280 gates and its estimated operating frequency is about 125Mhz at operating condition of $0.35{\mu}m$ CMOS technology. Because the processor can execute arccosine function within 7 clock cycles, it has about 17 MOPS(million arccos operations per second) execution rate and can be applicable to mobile OpenVG processor. And because of its flexible architecture, it can be applicable to the various transcendental functions such as exponential, trigonometric and logarithmic functions via replacement of ROM and minor hardware modification.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.2
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• pp.89-95
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• 2003

The study of this experiment is focused on labor saving of healthy rice seedling production using PSF (polypropylene spunbonded fabrics) as thermal protection material. Several factors such as different compositions of nursery soil and PSF materials were tested to produce healthy rice seedlings. The inner thermal protection material in PE film (polyethylene film) showed $0.9-1.7^{\circ}C$ higher than that of PSF 40-100 $\textrm{gm}^{-2}$. The light transmittance-ratio also showed similar trends. It is considered that the appropriate PSF material density was 40 gm$\textrm{gm}^{-2}$ in accordance with economic values and healthy rice seedlings. Plant height and dry weight according to various nursery soil showed the rang-es of 8.5-14.2cm and 5.5-10.0mg, respectively. In composition of nursery soil, artificial soil combined with paddy soil was effective in producing healthy seedling for rice seedling production. The total sugar content also showed the difference between PSF 40, 60 $\textrm{gm}^{-2}$ PE film (0.43-0.52mg FW $\textrm{g}^{-1}$) and PSF 80, 100 $\textrm{gm}^{-2}$ (0.28-0.35mg FW $\textrm{g}^{-1}$) and it showed the same tendency among varieties as well as various nursery soil. These results demonstrate that PSF 40 $\textrm{gm}^{-2}$ economically affordable, and can be recommended as thermal protection material for producing good healthy rice seedling.

• Koreanishche Zeitschrift fur Deutsche Sprachwissenschaft
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• v.10
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• pp.231-249
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• 2004

In dieser Albeit wird der Versuch unternommen, die Charakteristik der Rhetorik von Hitler zu analysieren. Im Abschnitt fwei wird die allgemeine Charakteristik der Hillers Reden beschrieben. Die Reden von Hitler sind Elemente eines verzweigten, flexiblen und inhaltlich nor schwer bestimmten Komplexes von Manipulations-und Machttechniken, die trotz seiner Tendenz zur Zentralisierung nur schwer als System zu fassen sind. Dies macht die Schwierigkeit jeder analytischen Erfassung aus. Seine Rede vermag im Bezug auf Logik und Argumentation wenig zu $\"{u}berzeugen$, aber or wirkte. In diesel Analyse wird die Rede von Hitler folgende Systematisierung $m\"{o}glich$: 1) Am Anfang weist Hitler auf die allgemeine Not hin. 2) Danach diffamiert or seine Gesner und provoziert starke Emotionen bei seinen $Zuh\"{o}rern$. 3) Zum $Schlu{\ss}$ $l\"{o}st$ Hitler bei den Deutschen eine starke Kampfbereitschaft aus und entwickelt vor semen $Zuh\"{o}rern$ seine Vision eines $bl\"{u}henden$, starken Deutschlands. 4) $Abschlie{\ss}end$ erkifirt Hitier seine $pers\"{o}nlichen$ ethischen Vorstellungen und seine sich daraus ergebenden Handlungen. Er liegt $gro{\ss}en$Wert darauf, die Ethik seine Handlungen zu $begr\"{u}nden$. Danach wird die Selbstdarstellung von Hitler beschrieben. $F\"{u}nf$ Realisierungsformen sind $daf\"{u}r$ relevant: a) $pers\"{o}nliche$ $Erz\"{a}hlungen$, b) Diffamierungen, c) sittlich wertvolle Ermahnungen, d) Taten und e) Ziele. Im Abschnitt drei wird die Rhetorik von Hitler gemacht. Hier handeit es sich um Ethos und Pathos. Hitler versuchte, die durch die Aktivierung zwischen ihm und Publikum vorhanden, $unbewu{\ss}ten$ psychischen Potentiate, die durch den Akt der Rede aktiviert werden, zu realisieren. Er hat jeder Sammlung nur gesagt, was sie $h\"{o}ren$ wollte, den wahren Sachverhalt nur auf $h\"{o}chst$ vordergryndige Weise.

• Reproductive and Developmental Biology
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• v.35 no.4
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• pp.449-455
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• 2011

Induced pluripotent stem (iPS) cells have been generated from mouse and human somatic cells by etopic expression of transcription factors. iPS cells are indistinguishable from ES cells in terms of morphology and stem cell marker expression. Moreover, mouse iPS cells give rise to chimeric mice that are competent for germline transmission. However, mice derived from iPS cells often develop tumors. Furthermore, the low efficiency of iPS cell generation is a big disadvantage for mechanistic studies. Nonviral plasmid.based vectors are free of many of the drawbacks that constrain viral vectors. The histone deacetylase inhibitor valproic acid (VPA) has been shown to improve the efficiency of mouse and human iPS cell generation, and vitamin C (Vc) accelerates gene expression changes and establishment of the fully reprogrammed state. The MEK inhibitor PD0325901 (Stemgent) has been shown to increase the efficiency of the reprogramming of human primary fibroblasts into iPS cells. In this report, we described the generation of mouse iPS cells devoid of exogenous DNA by the simple transient transfection of a nonviral vector carrying 2A-peptide-linked reprogramming factors. We used VPA, Vc, and the MEK inhibitor PD0325901 to increase the reprogramming efficiency. The reprogrammed somatic cells expressed pluripotency markers and formed EBs.