• Title/Summary/Keyword: Protein synthesis

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A Fibrin Matrix Promotes the Differentiation of EMSCs Isolated from Nasal Respiratory Mucosa to Myelinating Phenotypical Schwann-Like Cells

  • Chen, Qian;Zhang, Zhijian;Liu, Jinbo;He, Qinghua;Zhou, Yuepeng;Shao, Genbao;Sun, Xianglan;Cao, Xudong;Gong, Aihua;Jiang, Ping
    • Molecules and Cells
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    • v.38 no.3
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    • pp.221-228
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    • 2015
  • Because Schwann cells perform the triple tasks of myelination, axon guidance and neurotrophin synthesis, they are candidates for cell transplantation that might cure some types of nervous-system degenerative diseases or injuries. However, Schwann cells are difficult to obtain. As another option, ectomesenchymal stem cells (EMSCs) can be easily harvested from the nasal respiratory mucosa. Whether fibrin, an important transplantation vehicle, can improve the differentiation of EMSCs into Schwann-like cells (SLCs) deserves further research. EMSCs were isolated from rat nasal respiratory mucosa and were purified using anti-CD133 magnetic cell sorting. The purified cells strongly expressed HNK-1, nestin, $p75^{NTR}$, S-100, and vimentin. Using nuclear staining, the MTT assay and Western blotting analysis of the expression of cell-cycle markers, the proliferation rate of EMSCs on a fibrin matrix was found to be significantly higher than that of cells grown on a plastic surface but insignificantly lower than that of cells grown on fibronectin. Additionally, the EMSCs grown on the fibrin matrix expressed myelination-related molecules, including myelin basic protein (MBP), 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) and galactocerebrosides (GalCer), more strongly than did those grown on fibronectin or a plastic surface. Furthermore, the EMSCs grown on the fibrin matrix synthesized more neurotrophins compared with those grown on fibronectin or a plastic surface. The expression level of integrin in EMSCs grown on fibrin was similar to that of cells grown on fibronectin but was higher than that of cells grown on a plastic surface. These results demonstrated that fibrin not only promoted EMSC proliferation but also the differentiation of EMSCs into the SLCs. Our findings suggested that fibrin has great promise as a cell transplantation vehicle for the treatment of some types of nervous system diseases or injuries.

산란계 사료중 미역과 뱅코마이신이 산란계의 생산성에 미치는 영향

  • Choe, Do-Yeol;Im, Jin-Taek;Park, In-Gyeong;Choe, Jun-Yeong;Lee, Hye-Jeong;Lee, Beom-Gyu;Lee, Ho-Yeon;Park, Jae-U;Go, Tae-Song
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2005.11a
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    • pp.74-75
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    • 2005
  • In order to evaluate dietary brown seaweed and vancomycin on the performance, layers(Isa brown) were fed on basal diet and diets containing 2.0% of brown seaweed or 10ppm vancomycin. Brown seaweed diet significantly increased(p<0.01) nitrogen balance in layer, while excretion of uric acid nitrogen and metabolizable energy utilization were not different among diets. Layer consumed more the brown seaweed diet(p<0.05). Egg production were significantly different by diets but reduced(p<0.0001) with the experimental period passed. Layer fed brown seaweed diet gave thicker shell eggs, higher Haugh unit and higher egg white CuZnSOD activity compared with those in basal diet. Also, Brown seaweed diet increased MnSOD activity in erythrocyte cytosol and peroxidase in plasma, but decreased peroxide level in plasma, and increased proliferation of PBMC stimulated with PHA-P The result indicated that brown seaweed 2.0% diet in layer improved egg quality and performance due to increased protein synthesis which were related to regulation of antioxidant system and immune cell function in blood.

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Studies on the Biological Behaviors of Taxol Derivatives (Taxol 유도체들의 생물학적 거동에 관한 연구)

  • Awh, Ok-Doo;Yoo, Dae-Wung;Im, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.31 no.4
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    • pp.440-451
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    • 1997
  • This study was designed to prospect the $^{111}In$-labelled paclitaxel as tumor imaging agent. In order to provide a taxol molecule with a functional group which is able to chelate In-111, taxol-DTPA conjugate and 2'-hemisuccinyltaxol were synthesized by esterification of taxol at C-2'on C-13 carbon with DTPA anhydride and succinic anhydride, respectively. Synthesis yield of the taxol derivatives was 34% for taxol-DTPA and 80% for 2'-hemisuccinyltaxol. Cytotoxicity of the taxol derivatives were measured by MTT method toward cell lines HT29, B16, P388, and CT26. The cytotoxic activities of the taxol derivatives were maintained, although less active than taxol. Radiolabelling of the taxol derivatives were proceeded directly with $^{111}InCl_3$ or indirectly with $^{111}In$-citrate(ligand-exchange method). The ligand-exchange method was not suitable because some precipitates appeared during the reaction. On the contrary, by direct radiolabelling method, we were able to obtain taxol-DTPA-$^{111}In$ in 100% radiochemical yield. However, 2'-hemisuccinyltaxol was not labelled by both methods. Yield and radiochemical purity of the radiolabelled com-pound were determined by HPLC, paper chromatography and instant thin layer chromatography. Taxol-DTPA-$^{111}In$ was characterized to be hydrophilic by lipophilicity test, and nearly non-adhesive to HT29, B16, P388, and CT26 by cell binding affinity test. Binding affinity of the taxol-DTPA-$^{111}In$ complex to serum proteins was also examined by protein precipitation with 30% trichloroacetic acid. The results showed that 30% of the taxol-DTPA-$^{111}In$ complex binds with serum proteins.

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Targeting Analysis of Lumenal Proteins of Chloroplast of Wheat using Proteomic Techniques

  • Kamal, Abu Hena Mostafa;Kim, Da-Eun;Oh, Myoung-Won;Chung, Keun-Yook;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Uozumi, Nobuyuki;Choi, Jong-Soon;Cho, Kun;Woo, Sun-Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.05a
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    • pp.14-14
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    • 2010
  • Plastid proteomics are essential organelles present in virtually all cells in plants and green algae. Plastids are responsible for the synthesis and storage of key molecules required for the basic architecture and functions of plant cells. The proteome of plastid, and in particular of chloroplast, have received significant amounts of attention in recent years. Various fractionation and mass spectrometry (MS) techniques have been applied to catalogue the chloroplast proteome and its sub-organelles compartments. To better understanding the function of the lumenal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine-SDS-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, TMHMM, and TOPPRED) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.

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Cell cycle-dependent entry of Toxoplasma gondii into synchronized HL-60 cells (세포 주기 변화에 따른 Toxoplasmu gondii의 침투 양상)

  • 윤지혜;남호우
    • Parasites, Hosts and Diseases
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    • v.29 no.2
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    • pp.121-128
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    • 1991
  • The degree of attraction of Toxoplasma gondii to vertebrate cells varies with cell type and cell phase. Human promyelocytic leukemia cells, HL-60, were synchronized by double thymidine block method and co-cultured with Toxoplasma for 1 hr at each cell stage to investigate the cell cycle specific susceptibility of parasites to host cells. For 30 hr the average number of Texoplasma that invaded was a little changed except at 3 hr from G1/S phase boundary which concurred with the peak point of DNA synthesis. At 3 hr which is a relatively short interval compared to whole S phase, modification of cells by parasitic invasion was most remarkable. The number of Toxoplasma that penetrated was increased to more than sin times. The shape of the cells became sludgy and almost indiscernible by strong accessibility of parasites only for an hour of mfd-S phase. The same auctuation was also observed at the second peak of S phase but weakly. This suggests that there be surface molecules concerning with the attachment of Texoplasma to the host cells, which is expressed at special point of S phase. further studies on the specific protein or similar molecules related could be carried out using synchronized HL-60 cells.

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Anti-Proliferative Activity of OD78 Is Mediated through Cell Cycle Progression by Upregulation p27kip1 in Rat Aortic Vascular Smooth Muscle Cells

  • Tudev, Munkhtsetseg;Lim, Yong;Park, Eun-Seok;Kim, Won-Sik;Lim, Il-Ho;Kwak, Jae-Hwan;Jung, Jae-Kyung;Hong, Jin-Tae;Yoo, Hwan-Soo;Lee, Mi-Yea;Pyo, Myoung-Yun;Yun, Yeo-Pyo
    • Biomolecules & Therapeutics
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    • v.19 no.2
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    • pp.187-194
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    • 2011
  • Atherosclerosis and post-angiography restenosis are associated with intimal thickening and concomitant vascular smooth muscle cell (VSMC) proliferation. Obovatol, a major biphenolic component isolated from the Magnolia obovata leaf, is known to have anti-inflammatory and anti-tumor activities. The goal of the present study was to enhance the inhibitory effects of obovatol to improve its potential as a preventive or therapeutic agent in atherosclerosis and restenosis. Platelet-derived growth factor (PDGF)-BB-induced proliferation of rat aortic smooth muscle cells (RASMCs) was examined in the presence or absence of a newly synthesized obovatol derivative, OD78. The observed anti-proliferative effect of OD78 was further investigated by cell counting and [$^3H$]-thymidine incorporation assays. Treatment with 1-4 ${\mu}M$ OD78 dose-dependently inhibited the proliferation and DNA synthesis of 25 ng/ml PDGF-BB-stimulated RASMCs. Accordingly, OD78 blocked PDGF-BB-induced progression from the $G_0/G_1$ to S phase of the cell cycle in synchronized cells. OD78 decreased the expression levels of CDK4, cyclin E, and cyclin D1 proteins, as well as the phosphorylation of retinoblastoma protein and proliferating cell nuclear antigen; however, it did not change the CDK2 expression level. In addition, OD78 inhibited downregulation of the cyclin-dependent kinase inhibitor (CKI) $p27^{kip1}$. However, OD78 did not affect the CKI $p21^{cip1}$ or phosphorylation of early PDGF signaling pathway. These results suggest that OD78 may inhibit PDGF-BB-induced RASMC proliferation by perturbing cell cycle progression, potentially through $p27^{kip1}$ pathway activation. Consequently, OD78 may be developed as a potential anti-proliferative agent for the treatment of atherosclerosis and angioplasty restenosis.

Effect of Ground Corn Cob Replacement for Cassava Chip on Feed Intake, Rumen Fermentation and Urinary Derivatives in Swamp Buffaloes

  • Wanapat, M.;Pilajun, R.;Kang, S.;Setyaningsih, K.;Setyawan, A.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.8
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    • pp.1124-1131
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    • 2012
  • Four Thai - rumen fistulated male swamp buffaloes (Bubalus bubalis), about four years old with $400{\pm}20kg$ liveweight, were randomly assigned according to a $4{\times}4$ Latin square design to receive dietary treatments. The treatments were: ground corn cob (GCC) replacement for cassava chip (CC) in concentrate at 0% (T1); GCC replacement at 33% (T2); GCC replacement at 67% (T3); and GCC replacement at 100% (T4), respectively. During the experiment, concentrate was offered at 0.5% BW while 5% urea-treated rice straw was given at ad libitum. The result revealed that there was no effect of GCC replacement on DMI among treatments. In addition, digestibilities of DM, OM and CP were not different while aNDF linearly increased with an increasing level of GCC replacement. However, GCC replacement did not affect rumen fermentation such as ruminal pH, $NH_3$-N and VFA concentration; except C3 proportion which was the highest at 33% replacement while the lowest was at 100% replacement. All replacements of GCC resulted in similar protozoal and bacterial populations and microbial protein synthesis (MPS). Purine derivatives (PD) concentration in urine and PD to creatinine (PDC) index were varied with time of urination and among treatments at 0 to 8 and 8 to 16 h post feeding and higher values were shown among the GCC replacement groups. However at 16 to 24 h-post feeding, it was untraceable. In addition, creatinine concentration was similar among all treatments at every sampling time. Based on the above results, GCC can be used as an energy source for swamp buffalo fed with rice straw. Spot sampling of urine can be used for purine derivatives determination.

Rumen Parameters and Urea Kinetics in Goats and Sheep

  • Darlis, N. Abdullah;Halim, R.A.;Jalaludin, S.;Ho, Y.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.7
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    • pp.922-928
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    • 2000
  • The effects of animal species and supplements on rumen fluid characteristics, plasma urea-N (PUN) concentration, plasma urea-N pool size, urea-N degradation in the gut and urea-N net flux (urea-N synthesis rate) were studied in goats and sheep, with some minor differences detected. The animals were fed either chopped rice straw ad libitum+200 g soybean meal (SBM), or chopped rice straw ad libitum+190 g soybean meal+300 g sago meal (SBM+SM) for 14 days. The supplements were isonitrogenous (80 g crude protein/animal/d). [$^{14}C$]-urea was used as the marker for urea metabolism studies. Two animals from each species were fed either supplement in a cross-over design in two periods. The results showed that rumen pH was significantly (p<0.001) lower in animals fed SBM+SM than those fed SBM supplement. The ammonia concentrations of rumen fluid were significantly (p<0.01) higher in sheep (382.9 mg N/L) than goats (363.1 mg N/L) when fed SBM supplement but lower (282.5 mg N/L) than that of goats (311.0 mg N/L) when fed SBM+SM supplement. Total VFA concentrations were significantly (p<0.05) higher in animals fed SBM+SM supplement than those fed SBM supplement. Goats had significantly (p<0.01) higher molar proportions of acetate (79.1, 77.7%, respectively) than sheep (75.8, 74.0%, respectively) in both supplements. The molar proportion of acetate was significantly (p<0.05) higher, while that of butyrate lower in animals fed SBM supplement than those fed SBM+SM supplement. In animals fed SBM supplement, the molar proportion of propionate was significantly (p<0.01) higher in sheep (18.0%) than in goats (15.6%), but in animals fed SBM+SM, the molar proportion of butyrate was significantly (p<0.01) higher (9.6%) in sheep than in goats (7.2%). Plasma urea-N concentration, plasma urea-N pool size, urea-N degradation in the gut, urea-N net flux and the fraction of urea-C from the blood entering the rumen were not significantly different between goats and sheep fed either supplement. However, PUN concentration was significantly (p<0.05) lower in animals fed SBM+SM supplement (average of 13.8 mg N/100 ml) than in those fed SBM supplement (average of 16.5 mg N/100 ml). The urea net flux was significantly (p<0.05) higher in goats (average of 14.5 g N/d) than sheep (average of 12.9 g N/d), and animals fed SBM supplement showed higher (average of 14.9 g N/d) urea net flux than animals fed SBM+SM supplement (average of 12.9 g N/d). A significant (p<0.05) positive correlation was observed between urea-N net flux and urea-N degradation; urea-N net flux and pool size; urea-N net flux and urea excretion in the urine; and PUN and rumen ammonia in goats. While in sheep, significant (p<0.05) positive correlation was observed between urea-N net flux and urea excretion in the urine; and PUN and rumen ammonia.

Antimelanogenic Effect of Purpurogallin in Murine Melanoma Cells (마우스 흑색종세포에서 Purpurogallin의 멜라닌 생성 억제 효과)

  • Kim, Han-Hyuk;Kim, Tae Hoon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.12
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    • pp.1905-1911
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    • 2015
  • Melanin is one of the most important factors affecting skin color. Melanogenesis is the bioprocess of melanin production by melanocytes in the skin and hair follicles and is mediated by several enzymes, such as tyrosinase, tyrosinase related protein (TRP)-1, and TRP-2. Convenient enzymatic transformation of the simple phenol pyrogallol with polyphenol oxidase originating from pear to an oxidative product, purpurogallin, was efficient. The structure of the pyrogallol oxidation product was identified on the basis of spectroscopic methods. The biotransformation product purpurogallin showed significant inhibitory effects against both melanin synthesis and tyrosinase activity in a dose-dependent manner in B16 melanoma cells. In addition, purpurogallin significantly attenuated melanin production by inhibiting TRP-1, and TRP-2 expression through modulation of their corresponding transcription factors, and microphthalamia- associated transcription factor in B16 cells. Consequently, purpurogallin derived from convenient enzymatic transformation of pyrogallol might be a beneficial material for reducing skin hyperpigmentation.

Interactions between Biosynthetic Pathway and Productivity of IAA in Some Rhizobacteria (근권에서 분리한 세균의 IAA 생합성 경로와 IAA 생성능과의 관계)

  • Kim, Woon-Jin;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.48 no.1
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    • pp.1-7
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    • 2012
  • This study explores the interaction between the production of indole-3-acetic acid (IAA), a typical phytohormone auxin and the role of IAA biosynthetic pathways in each IAA producing rhizobacterial strain. The bacterial strains were isolated from rhizosphere of wild plants and identified as Acinetobacter guillouiae SW5, Bacillus thuringiensis SW17, Rhodococcus equi SW9, and Lysinibacillus fusiformis SW13. A. guillouiae SW5 exhibited the highest production of IAA using tryptophan-dependent pathways among the 4 strains. When indole-3-acetamide (IAM) was added, Rhodococcus equi SW9 showed the highest IAA production of $3824{\mu}g/mg$ protein using amidase activity. A. guillouiae SW5 also showed the highest production of IAA using two pathways with indole-3-acetonitrile (IAN), and its nitrile hydratase activity might be higher than nitrilase. B. thuringiensis SW17 showed the lowest IAA production, and most of IAA might be produced by the amidase activity, although the nitrilase activity was the highest among 4 strains. The roles of nitrile converting enzymes were relatively similar in IAA synthesis by Lysinibacillus fusiformis SW13. Tryptophan-independent pathway of IAA production was utilized by only A. guillouiae SW5.