• Title/Summary/Keyword: $C_5$-precursor

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Effect of Parthenogenetic Mouse Embryonic Stem Cell (PmES) in the Mouse Model of Huntington′s Disease

  • 이창현;김용식;이영재;김은영;길광수;정길생;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.80-80
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    • 2003
  • Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder characterized by motor, cognitive, and psychiatric symptoms, accompanied by marked cell death in the striatum and cortex. Stereotaxic injection of quinolinic acid (QA) into striatum results in a degeneration of GABAergic neurons and exhibits abnormal motor behaviors typical of the illness. The objective of this study was carried out to obtain basic information about whether parthenogenetic mouse embryonic stem (PmES) cells are suitable for cell replacement therapy of HD. To establish PmES cell lines, hybrid F1 (C57BL/6xCBA/N) mouse oocytes were treated with 7% ethanol for 5 min and cytochalasin-B for 4 hr to initiate spontaneous cleavage. Thus established PmES cells were induced to differentiate using bFGF (20ng/ml) followed by selection of neuronal precursor cells for 8 days in N2 medium. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days, then a final differentiation step in N2 medium for 7 days. To establish recipient animal models of HD, young adult mice (7 weeks age ICR mice) were lesioned unilaterally with a stereotaxic injection of QA (60 nM) into the striatum and the rotational behavior of the animals was tested using apomorphine (0.1mg/kg, IP) 7 days after the induction of lesion. Animals rotating more than 120 turns per hour were selected and the differentiated PmES cells (1$\times$10$^4$cells/ul) were implanted into striatum. Four weeks after the graft, immunohistochemical studies revealed the presence of cells reactive to anti-NeuN antibody. However, only a slight improvement of motor behavior was observed. By Nissl staining, cell mass resembling tumor was found at the graft site and near cortex which may explain the slight behavioral improvement. Detailed experiment on cell viability, differentiation and migration explanted in vivo is currently being studied.

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An Antifungal Antibiotic Purified from Bacillus megaterium KL39, a Biocontrol Agent of Red-Pepper Phytophthora-Blight Disease

  • JUNG HEE KYOUNG;KIM SANG-DAL
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1001-1010
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    • 2005
  • Bacillus megaterium KL39, an antibiotic-producing plant growth promoting rhizobacterium (PGPR), was selected from soil. The antifungal antibiotic, denoted KL39, was purified from culture filtrate by column chromatography using Dion HP-20, Silica gel, Sephadex LH-20, and prep-HPLC. Thin layer chromatography, employing the solvent system of ethanol:ammonia:water=8:1:1, showed the $R_{f}$. value of 0.32. The antibiotic KL39 showed a negative reaction with ninhydrin solution, positive with iodine vapor, and also positive with Ehrlich reagent. It was soluble in methanol, ethanol, butanol, and acetonitrile, but insoluble in chloroform, toluene, hexane, ethyl ether, or acetone. Its UV spectrum had the maximum absorption at 208 nm. Amino acid composition, FAB-mass, $^{1}H-NMR,\;^{13}C-NMR$, and atomic analyses showed that the antibiotic KL39 (MW=1,071) has a structure very similar to iturin E. The antibiotic KL39 has a broad antifungal spectrum against a variety of plant pathogenic fungi including Rhizoctonia solani, Pyricularia oryzae, Monilinia froeticola, Botrytis cinenea, Altenaria kikuchiana, Fusarium oxysporum, and F. solani. An MIC value of $10\;{\mu}g/ml$ was determined for Phytophthora capsici. Macromolecular incorporation studies with P. capsici using radioactive [$^{3}H-adenine$] as the precursor, indicated that the antibiotic KL39 strongly inhibits the DNA biosynthesis of the fungal cell. Microscopic observation of the antifungal action showed abnormal hyphal swelling of P. capsici. The purified antibiotic KL39 was very effective for the biocontrol of in vivo Phytophthora-blight disease of pepper.

Physical Properties and Morphology of Carbon Nanotubes Prepared by Thermal and Plasma CVD of Acetylene (아세틸렌의 열 및 플라즈마 CVD법으로 제조한 탄소나노튜브의 물성과 구조적 특성)

  • Kim, Myung-Chan;Moon, Seung-Hwan;Lim, Jae-Seok;Hahm, Hyun-Sik;Kim, Myung-Soo
    • Journal of the Korean Applied Science and Technology
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    • v.21 no.2
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    • pp.174-181
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    • 2004
  • Multi-walled carbon nanotubes (CNTs) were prepared by thermal chemical vapor deposition (CVD) and microwave plasma chemical vapor deposition (MPCVD) using various combination of binary catalysts with four transition metals such as Fe, Co, Cu, and Ni. In the preparation of CNTs from acetylene precursor by thermal CVD, the CNTs with very high yield of 43.6 % was produced over $Fe-Co/Al_2O_3$. The highest yield of CNTs was obtained with the catalyst reduced for 3 hr and the yield was decreased with increasing reduction time to 5 hr, due to the formation of $FeAl_2O_4$ metal-aluminate. On the other hand, the CNTs prepared by acethylene plasma CVD had more straight, smaller diameter, and larger aspect ratio(L/D) than those prepared by thermal CVD, although their yield had lower value of 27.7%. The degree of graphitization of CNTs measured by $I_d/I_g$ value and thermal degradation temperature were 1.04 and $602^{\circ}C$, respectively.

Study on Myocardial Protective Effect of L-arginine (L-Arginine의 흰쥐 적출심근보호 효과에 관한 연구)

  • 이인성;이헌재
    • Journal of Chest Surgery
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    • v.29 no.10
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    • pp.1076-1080
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    • 1996
  • The results about the myocardial protection of recta of the nitric oxide precursor L-arginine upon reperrusion injury after ischemia are diverse. These diversities may be model dependent. Experiments were designed and performed to investigate myocardial protection effects according to the concentration of L-arginine. The Isolated rat hearts were subjected in a 30 minutes oi normothermic ischemia and reperfused for 30 minutes with reperfusate containing 0, 1, 2, 3, 4 moil L-arginine. After 30 minutes of reperfusion, group with 1 and 2 mM/L L-arginine showed a trend of better recovery in left ventricular systolic function(left ventricular developed pressure, positive maximum dpfdt), diastolic function(negative maximum dpfdt) and coronary flow compared to control group(reperfusate no L-arginine). Recovery was impaired with a higher concentration, and at 4 moil L-arginine r covery was worse than control(p (0, 05). These results suggest that optimal concentration of L-arginine Is Important or the recovery of myocardial and endothelial function after ischemia and reperfusion.

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Determination of isoquinoline alkaloids by UPLC-ESI-Q-TOF MS: Application to Chelidonium majus L.

  • Jeong, Won Tae;Lim, Heung Bin
    • Analytical Science and Technology
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    • v.30 no.6
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    • pp.379-389
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    • 2017
  • In this study, we set up an analytical method that can be used for rapid and accurate determination of representative isoquinoline alkaloids in medicinal plants using UPLC-ESI-Q-TOF MS (ultra pressure liquid chromatography-electrospray ionization-quadrupole-time-of-flight mass spectrometry). The compounds were eluted on a C18 column with 0.1 % formic acid and acetonitrile, and separated with good resolution within 13 min. Each of the separated components was characterized by precursor ions (generated by ESI-Q-TOF) and fragment ions (produced by collision-induced dissociation, CID), which were used as a reliable database. We also performed method validation: analytes showed excellent linearity ($R^2$, 0.9971-0.9996), LOD (5-25 ng/mL), LOQ (17-82 ng/mL), accuracy (91.6-97.4 %) as well as intra- and inter-day precisions (RSD, 1.8-3.2 %). In the analysis of Chelidonium majus L., magnoflorine, coptisine, sanguinarine, berberine and palmatine were detected by matching retention times and characteristic fragment ion patterns of reference standards. We also confirmed that, among the quantified components, coptisine was present in the highest quantity. Furthermore, alkaloid profiling was carried out by analyzing the fragment ion patterns corresponding to peaks of unknown components. In this manner, protopine, chelidonine, stylopine, dihydroberberine, canadine, and nitidine were tentatively identified. We also proposed the molecular structure of the fragment ions that appear in the mass spectrum. Therefore, we concluded that our suggested method for the determination of major isoquinoline alkaloids by UPLC-Q-TOF can be useful not only for quality control, but also for rapid and accurate investigation of phytochemical constituents of medicinal plants.

Superhydrophobic nanostructured non-woven fabric using plasma modification

  • Shin, Bong-Su;Lee, Kwang-Reoul;Kim, Ho-Young;Moon, Myoung-Woon
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.320-320
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    • 2011
  • We describe fabrication of superhydrophobic surface on non-woven fabric (NWF) having nano-hairy structures and a hydrophobic surface coating. Oxygen plasma was irradiated on NWF for nano-texuring and a precursor of HMDSO (Hexamethydisiloxane) was introduced as a surface chemical modification for obtaining superhydrophobicity using 13.56 MHz radio frequency-Plasma Enhanced Chemical Vapor Deposition (rf-PECVD). O2 plasma treatment time was varied from 1 min to 60 min at a bias voltage of 400V, which fabricated pillar-like structures with diameter of 30 nm and height of 150 nm on NWF. Subsequently, hydrophobic coating using hexamethyldisiloxane vapor was deposited with 10 nm thickness on NWF substrate at a bias voltage of 400 V. We evaluate superhydrophobicity of the modified NWF with sessile drop using goniometer and high speed camera, in which aspect ratio of nanohairy structures, contact angle and contact angle hysteresis of the surfaces were measured. With the increase of aspect ratio, the wetting angle increased from $103^{\circ}$ to $163^{\circ}$, and the contact angle hysteresis decreased dramatically below $5^{\circ}$. In addition, we had conducted experiment for nucleation and condensation of water via E-SEM. During increasing vapor pressure inside E-SEM from 3.7 Torr to over 6 Torr which is beyond saturation point at $2^{\circ}C$, we observed condensation of water droplet on the superhydropobic NWF. While the condensation of water on oxygen plasma treated NWF (superhydrophilic) occurred easily and rapidly, superhydrophobic NWF which was fabricated by oxygen and HMDSO was hardly wet even under supersaturation condition. From the result of wetting experiment and water condensation via E-SEM, it is confirmed that superhydrophobic NWF shows the grate water repellent abilities.

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Identification and Functional Analysis of Mating Type Loci in the Pleurotus eryngii

  • Ryu, Jae San;Kim, Min-Keun;Park, Bokyung;Ali, Asjad;Joung, Wan-Kyu
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.35-35
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    • 2015
  • Pleurotus eryngii has recently become a major cultivated mushroom; it uses tetrapolar heterothallism as a part of its reproductive process. Sexual development progresses only when the A and B mating types are compatible. Such mating incompatibility occasionally limits the efficiency of breeding programs in which crossing within loci-shared strains or backcrossing strategies are employed. Therefore, understanding the mating system in edible mushroom fungi will help provide a short cut in the development of new strains. We isolated and identified pheromone and receptor genes in the B3 locus of P. eryngii and performed a functional analysis of the genes in the mating process by transformation. A genomic DNA library was constructed to map the entire mating-type locus. The B3 locus was found to contain four pheromone precursor genes and four receptor genes. Remarkably, receptor PESTE3.3.1 has just 34 amino acid residues in its C-terminal cytoplasmic region; therefore, it seems likely to be a receptor-like gene. Real-time quantitative RT-PCR (real-time qRT-PCR) revealed that most pheromone and receptor genes showed significantly higher expression in monokaryotic cells than dikaryotic cells. The pheromone genes PEphb3.1 and PEphb3.3 and the receptor gene PESTE3.3.1 were transformed into P5 (A3B4). The transformants were mated with a tester strain (A4B4), and the progeny showed clamp connections and a normal fruiting body, which indicates the proposed role of these genes in mating and fruiting processes. This result also confirms that PESTE3.3.1 is a receptor gene. In this study, we identified pheromone and receptor genes in the B3 locus of P. eryngii and found that some of those genes appear to play a role in the mating and fruiting processes. These results might help elucidate the mechanism of fruiting differentiation and improve breeding efficiency.

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Size-controlled Chevrel Mo6S8 as Cathode Material for Mg Rechargeable Battery

  • Ryu, Anna;Park, Min-Sik;Cho, Woosuk;Kim, Jeom-Soo;Kim, Young-Jun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.10
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    • pp.3033-3038
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    • 2013
  • Nanoscale Chevrel $Mo_6S_8$ powders are synthesized by molten salt synthesis. Synthesized $Mo_6S_8$ powders have different mean particle sizes which are dependent on a ratio of salt to precursor. The particle sizes of $Mo_6S_8$ powders changes along with the ratio increase. $Mo_6S_8$ (6:1) demonstrates the best electrochemical characteristics among the synthesized $Mo_6S_8$ powders although the $Mo_6S_8$ (4:1) has the smallest particle size. $Mo_6S_8$ (6:1) shows a reversible capacity of 83.9 $mAhg^{-1}$, which is 27.5% and 33% improved value over $Mo_6S_8$ (2:1) and $Mo_6S_8$ (4:1) at a current density of 0.2C, respectively. The superior electrochemical properties of $Mo_6S_8$ (6:1) are attributed to the balanced particle size which provides proper contact area with electrolyte and the shortened $Mg^{2+}$ diffusion length. The $Mo_6S_8$ (4:1) has the smallest particle size but further reduction of particle size from $Mo_6S_8$ (6:1) is not advantageous.

Biochemical Characterization of a GDSL-Motif Esterase from Bacillus sp. K91 with a New Putative Catalytic Mechanism

  • Ding, Junmei;Yu, Tingting;Liang, Lianming;Xie, Zhenrong;Yang, Yunjuan;Zhou, Junpei;Xu, Bo;Li, Junjun;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1551-1558
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    • 2014
  • The esterase gene Est8 from the thermophilic bacterium Bacillus sp. K91 was cloned and expressed in Escherichia coli. The monomeric enzyme exhibited a theoretical molecular mass of 24.5 kDa and an optimal activity around $50^{\circ}C$ at pH 9.0. A model of Est8 was constructed using a hypothetical YxiM precursor structure (2O14_A) from Bacillus subtilis as template. The structure showed an ${\alpha}/{\beta}$-hydrolase fold and indicated the presence of a typical catalytic triad consisting of Ser-11, Asp-182, and His-185, which were investigated by site-directed replacements coupled with kinetic characterization. Asp-182 and His-185 residues were more critical than the Ser-11 residue in the catalytic activity of Est8. A comparison of the amino acid sequence showed that Est8 could be grouped into the GDSL family and further classified as an SGNH hydrolase. Est8 is a new member of the SGNH hydrolase subfamily and may employ a different catalytic mechanism.

Expression of manB Gene from Escherichia coli in Lactococcus lactis and Characterization of Its Bifunctional Enzyme, Phosphomannomutase

  • Li, Ling;Kim, Seul Ah;Fang, Ruosi;Han, Nam Soo
    • Journal of Microbiology and Biotechnology
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    • v.28 no.8
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    • pp.1293-1298
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    • 2018
  • Phosphomannomutase (ManB) converts mannose-6-phosphate (M-6-P) to mannose-1-phosphate (M-1-P), which is a key metabolic precursor for the production of GDP-D-mannose used for production of glycoconjugates and post-translational modification of proteins. The aim of this study was to express the manB gene from Escherichia coli in Lactococcus lactis subsp. cremoris NZ9000 and to characterize the encoded enzyme. The manB gene from E. coli K12, of 1,371 bp and encoding 457 amino acids (52 kDa), was cloned and overexpressed in L. lactis NZ9000 using the nisin-controlled expression system. The enzyme was purified by Ni-NTA column chromatography and exhibited a specific activity of 5.34 units/mg, significantly higher than that of other previously reported ManB enzymes. The pH and temperature optima were 8.0 and $50^{\circ}C$, respectively. Interestingly, the ManB used in this study had two substrate specificity for both mannose-1-phosphate and glucose-1-phosphate, and the specific activity for glucose-1-phosphate was 3.76 units/mg showing 70% relative activity to that of mannose-1-phosphate. This is the first study on heterologous expression and characterization of ManB in lactic acid bacteria. The ManB expression system constructed in this study canbe used to synthesize rare sugars or glycoconjugates.