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Partial Purification of Protein X from the Pyruvate Dehydrogenase Complex of Bovine Kidney

  • ;;;;Richard L. Veech
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.260-260
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    • 1994
  • Mammalian pyruvate dehydrogenase complex(PDC) enzyme consists of multiple oopies of three major oligomeric enzymes-El, E2 E3. And protein X is one of the enzymatic constituents which is tightly bound to E2 subunit This complex enzyme is responsible for the oxidative decarboxylation of pyruvate producing of acetyl CoA which is a key intermediate for the entry of carbohydrates into the TCA cycle for its complete metabolic conversion to CO$_2$. And the overall activity of the complex enzyme is regulated via covalent nodification of El subunit by a El specific phosphatase ad kinase. Protein X has lipoyl moiety that undergoes reduction and acetylation during ezymatic reaction and has been known h be involved in the binding of E3 subunit to E2 core and in the regulatory activity of kinase. The purification of protein X has not been achieved majorly because of its tight binding to E2 subunit The E2-protein X subcomplex was obtained by the established methods and the detachment of protein X from E2 was accomplished in the 0.1M borate buffer containing 150mM NaCl. During the storage of the subcomplex in frozen state at -70$^{\circ}C$, the E2 subunit was precipitated and the dissociated protein X was obtained by cntrifegation into the supernatant The verification of protein X was accomplished by (1)the migration on SDS-PAGE, (2)acetylation by 〔2$\^$-l4/C〕 pyruvate, and (3)internal amino acid sequence analysis of tryptic digested enzyme.

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Phospholipid Polymer, 2-Methacryloyloxyethyl Phosphorylcholine and Its Skin Barrier Function

  • LeeCho, Ae-Ri
    • Archives of Pharmacal Research
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    • v.27 no.11
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    • pp.1177-1182
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    • 2004
  • The effect of poly[2-methacryloyloxyethyl phosphorylcholine] (pMPC) on the skin permeation property was investigated by performing in vitro skin permeation study of a model drug, nicotinic acid (NA). Effect of pMPC polymer in donor solution on skin permeation rates was evaluated using side-by-side diffusion cells. Also, the structural alterations in the stratum corneum (SC), inter-lamellar bilayer (ILB) and dermis layers in pMPC-treated and -untreated skin sections were investigated with transmission electron microscopy (TEM). The permeation profile of NA without pMPC in donor solution showed biphasic mode: initial $1^{st} phase and 2^{nd}$ hydration phase. The sudden, more than 10-fold increase in flux from the initial steady state (43.5 $\mu g/cm^2$/hr) to the $2^{nd}$ hydration phase (457.3 $\mu g/cm^2$/hr) suggests the disruption of skin barrier function due to extensive hydration. The permeation profile of NA with 3% pMPC in the donor solution showed monophasic pattern: the steady state flux (10.9 $\mu g/cm^2$/hr) without abrupt increase of the flux. The degree of NA permeation rate decreased in a concentration-dependent manner of pMPC. TEM of skin equilibrated with water or 2% pMPC for 12 h showed that corneocytes are still cohesive and epidermis is tightly bound to dermis in 2% pMPC-treated skin, while wider separation between corneocytes and focal dilations in inter-cellular spaces were observed in water-treated skin. This result suggests that pMPC could protect the barrier property of the stratum corneum by preventing the disruption of ILB structure caused by extensive skin hydration during skin permeation study.

Studies on the Application of Starch for paper surface sizing(III) - The influence of surface sizing treatment with starch on the quality of uncoated printing paper - (종이 표면 사이즈 프레스용 전분의 적용에 관한 연구 -표면 사이즈용 전분이 백상지 품질에 미치는 영향 -)

  • 윤지영;이용규
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.34 no.2
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    • pp.1-12
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    • 2002
  • Starch dissolved in paper-mill wastes, either as a result of poor retention on the paper web or recycling of surface-treated broke, was a major pollutant Laboratory tests were performed by using different kinds of starch as a surface treatment. It was concluded that the use of cationic starch can positively affect the level of starch dissolved in liquid effluents. When cationically modified starches were used for surface sizing, the starch was tightly bound to the paper fibers, it was not removed during the repulping of broke. The result of mill trial in fine paper manufacture for the application of low-viscosity cationic starches used in size press reduced COD load in the effluents and increased One Pass Retention. It had been found that when cationic starch used as a surface sizing agent, more starch was retained on or near the surface of the sheet than with conventional oxidized starches. Thus surface strengths and quality were improved. In addition it is possible to maintain the desired level of starch penetration into the fiber net and improve porosity, opacity and brightness. In contrast, in most cases, dusting problems are notably eliminated. Cationic surface sized starch improved black and color ink-jet print quality in terms of feathering and optical density of the print image. These improved properties were believed to be due to a combination of fiber bonding and surface orientation more uniform starch concentration on the paper surface was resulted. Moreover cationic charges in the paper surface lend themselves excellently to fix ink jet ink anionic in nature.

Tas13D Inhibits Growth of SMMC-7721 Cell via Suppression VEGF and EGF Expression

  • He, Huai-Zhen;Wang, Nan;Zhang, Jie;Zheng, Lei;Zhang, Yan-Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2009-2014
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    • 2012
  • Objective: Taspine, isolated from Radix et Rhizoma Leonticis has demosntrated potential proctiective effects against cancer. Tas13D, a novel taspine derivative synthetized by structure-based drug design, have been shown to possess interesting biological and pharmacological activities. The current study was designed to evaluate its antiproliferative activity and underlying mechanisms. Methods: Antiproliferative activity of tas13D was evaluated by xenograft in athymic mice in vivo, and by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and cell migration assays with human liver cancer (SMMC-7721) cell lines in vitro. Docking between tas13D and VEGFR and EGFR was studied by with a Sybyl/Surflex module. VEGF and EGF and their receptor expression was determined by ELISA and real-time PCR methods, respectively. Results: Our present study showed that tas13D inhibited SMMC-7721 xenograft tumor growth, bound tightly with the active site of kinase domains of EGFR and VEGFR, and reduced SMMC-7721 cell proliferation (IC=34.7 ${\mu}mol/L$) and migration compared to negative controls. VEGF and EGF mRNAs were significantly reduced by tas13D treatment in a dose-dependent manner, along with VEGF and EGF production. Conclusion: The obtained results suggest that tas13D inhibits tumor growth and cell proliferation by inhibiting cell migration, downregulating mRNA expression of VEGF and EGF, and decreasing angiogenic factor production. Tas13D deserves further consideration as a chemotherapeutic agent.

Epithelial-mesenchymal Transition and Its Role in the Pathogenesis of Colorectal Cancer

  • Zhu, Qing-Chao;Gao, Ren-Yuan;Wu, Wen;Qin, Huan-Long
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.2689-2698
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    • 2013
  • Epithelial-to-mesenchymal transition (EMT) is a collection of events that allows the conversion of adherent epithelial cells, tightly bound to each other within an organized tissue, into independent fibroblastic cells possessing migratory properties and the ability to invade the extracellular matrix. EMT contributes to the complex architecture of the embryo by permitting the progression of embryogenesis from a simple single-cell layer epithelium to a complex three-dimensional organism composed of both epithelial and mesenchymal cells. However, in most tissues EMT is a developmentally restricted process and fully differentiated epithelia typically maintain their epithelial phenotype. Recently, elements of EMT, specially the loss of epithelial markers and the gain of mesenchymal markers, have been observed in pathological states, including epithelial cancers. Increasing evidence has confirmed its presence in human colon during colorectal carcinogenesis. In general, chronic inflammation is considered to be one of the causes of many human cancers including colorectal cancer(CRC). Accordingly, epidemiologic and clinical studies indicate that patients affected by ulcerative colitis and Crohn's disease, the two major forms of inflammatory bowel disease, have an increased risk of developing CRC. A large body of evidence supports roles for the SMAD/STAT3 signaling pathway, the NF-kB pathway, the Ras-mitogenactivated protein kinase/Snail/Slug and microRNAs in the development of colorectal cancers via epithelial-tomesenchymal transition. Thus, EMT appears to be closely involved in the pathogenesis of colorectal cancer, and analysis refered to it can yield novel targets for therapy.

Quality of steak restructured from beef trimmings containing microbial transglutaminase and impacted by freezing and grading by fat level

  • Sorapukdee, Supaluk;Tangwatcharin, Pussadee
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.129-137
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    • 2018
  • Objective: The objective of this research was to evaluate the physico-chemical, microbiological and sensorial qualities of restructured steaks processed from beef trimmings (grade I and II) and frozen beef (fresh beef as control and frozen beef). Methods: Beef trimmings from commercial butcher were collected, designated into 4 treatments differing in beef trimmings grade and freezing, processed into restructured steaks with 1% microbial transglutaminase and then analyzed for product quality. Results: The results showed that all meat from different groups could be tightly bound together via cross-linking of myosin heavy chain and actin as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Microbial counts of psychrotrophic and mesophilic bacteria were not affected by treatments (p>0.05), and no detectable of thermophilic bacteria were found. Regarding effect of beef trimmings grade, steaks made from beef trimmings grade II (16.03% fat) showed some superior sensorial qualities including higher tenderness score (p<0.05) and tendency for higher scores of juiciness and overall acceptability (p<0.07) than those made from beef trimmings grade I (2.15% fat). Moreover, a hardness value from texture profile analysis was lower in steaks processed from beef trimmings grade II than those made from grade I (p<0.05). Although some inferior qualities in terms of cooking loss and discoloration after cooking were higher in steaks made from beef trimmings grade II than those made from beef trimmings grade I (p<0.05), these differences did not affect the sensory evaluation. Frozen beef improved the soft texture and resulted in effective meat binding as considered by higher cohesiveness and springiness of the raw restructured product as compared to fresh beef (p<0.05). Conclusion: The results indicated the most suitable raw beef for producing restructured steaks without detrimental effect on product quality was beef trimmings grade II containing up to 17% fat which positively affected the sensory quality and that frozen beef trimmings increased tenderness and meat binding of restructured beef steaks.

Thermoelectric Composites Based on Carbon Nanotubes and Micro Glass Bubbles (탄소나노튜브 및 마이크로 글래스 버블 기반 열전 복합재)

  • Kang, Gu-Hyeok;Seong, Kwangwon;Kim, Myungsoo;Kim, In Guk;Bang, In Cheol;Park, Hyung Wook;Park, Young-Bin
    • Composites Research
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    • v.28 no.2
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    • pp.70-74
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    • 2015
  • In this paper, carbon nanotubes (CNTs) and micro glass bubbles (GBs) have been incorporated into a polyamide6 (PA6) matrix to impart thermoelectric properties. The spaces created in the matrix by GBs allows the formation of "segregated" CNT network. The tightly bound CNT network, if controlled properly, can serve as a conductive path for electron transport, while prohibiting phonon transport, which would provide an ideal configuration for thermoelectric applications. The CNTs and GBs were dispersed in a nylon-formic acid solution using horn sonication followed by coagulation in deionized water, and nanocomposite panels were fabricated using a hot press. The performance of nanocomposite panels was evaluated from thermal and electrical conductivities and Seebeck coefficient, and a thermoelectric figure of merit as high as 0.016 was achieved.

Solution Dynamics Studies for the Lck SH2 Domain Complexed with Peptide and Peptide-Free Forms

  • Yoon, Jeong-Hyeok;Chi, Myung-Whan;Yoon, Chang-No;Park, Jongsei
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.81-81
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    • 1995
  • It is well known that Src Homology 2(SH2) domain in many intracellular signal transduction proteins is very important. The domain has about 100 amino acid residues and bind phosphotyrosine-containing peptide with high affinity and specificity. Lck SH2 domain is a Src-like, lymphocyte-specific tyrosine kinase. An 11-residue phosphopeptide derived from the hamster polvoma middle-T antigen, EPQp YEEIPIYL, binds with an 1 nM dissociation constant to Lck SH2 domain. And it is known that the phosphotyrosine and isoleucine residues of the peptide are tightly bound by two well-defined pockets on Lck SH2 domain's surface. To investigate the conformational changes during complexation of SH2 domain with phosphopeptide we have performed the molecular dynamics simulation for Lck SH2 domain with peptide and peptide-free form at look in aqueous solution. More than 3000 water molecules were incorporated to solvate Lck SH2 domain and peptide. Periodic boundary condition has been applied in molecular dynamics simulation. Data analysis with the results of that simulation shows that the phosphopeptide makes primary interaction with the Lck SH2 domain at six central residues, The comparison of the complexed and uncomplexed SH2 domain structures in solution has revealed only relatively small change. But the hydrophilic and hydrophobic pockets in the protein surface show the conformational changes in spite of the small structural difference between the complex and peptide-free forms.

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On the Possible Fusion-Promoting Factor Secreted from Cultured Myoblasts (培養 鷄胚 筋原細胞로부터 분비된 細胞融合 촉진 물질에 관한 연구)

  • Park, Hye-Gyeong;Park, Young-Chul;Lee, Chung-Choo;Ha, Doo-Bong
    • The Korean Journal of Zoology
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    • v.29 no.4
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    • pp.294-306
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    • 1986
  • In order to find out whether myoblast cells release into the culture medium any substances that induce or promote the fusion of myoblasts, chick embryonic myoblasts were cultured and the cultured medium (muscle-conditioned medium, MCM) was collected. The MCM was then added to the newly cultured myoblasts to examine if it has fusion-promoting activity. The MCM was also analyzed for its protein content before and after its addition to the second culture. The MCM apparently showed fusion-promoting activity when applied to unfused young myoblasts, suggesting that it contained substances that promote the fusion and that had been released from cells fo the previous culture. Analysis of proteins in the myoblasts and in the MCM suggested that the released protein was absorbed by or tightly bound to myoblasts of the second culture. One of the released proteins of about 175 kilodalton was degraded to a polypeptide of approximately 145 kilodalton, which appeared to act upon the membrane proteins of unfused myoblasts so as to stimulate their membrane to fuse with neighboring cells.

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A Formal Framework for Context-Aware System Modeling (상황인식 시스템 모델링을 위한 정형화 프레임워크)

  • Cho, Eun-Sun;Min, Young-Mok
    • Journal of the Institute of Electronics Engineers of Korea CI
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    • v.46 no.2
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    • pp.114-123
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    • 2009
  • Context-aware systems are reactive computing systems, aware of external context. Considering various sensors and actuators connected, application programming on top of such systems is known to be much more complex than in contentional ones. This paper suggests a formal framework for context-aware systems, by extracting their common properties. That makes a useful birds-eye view for the behaviors of a whole complex system, as a base for a convenient developing environment and systematic analysis. In this framework, reactive-ness is handled by event-condition-action rules and global state-transitions, which are essential in a lot of context-aware applications. In addition, behaviors of each elementary device are modelled with its own state-transitions, and tightly bound to the entire task.