• Title/Summary/Keyword: Molecular structures

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Structural Analysis of the Antitumor Active Exo-polysaccharide Produced by Submerged Cultivation of Ganoderma lucidum Mycelium (영지(Ganoderma lucidum) 균사체의 액체배양에 의한 세포외 항암활성 다당류의 구조분석)

  • Lee, Shin-Young;Kang, Tae-Su
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.76-81
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    • 1999
  • Exo-polysaccharide obtained from the submerged cultivation of Ganoderma lucidum mycelium was fractionated. The structural analysis of the acidic exo-polysaccharide fraction (BWS-DA-GI), showing high antitumor activity, was carried out and compared to the mycelial acidic fraction (MWS-DA-GI). The major sugar constituents of the fraction of BWS-DA-GI were glucose, galactose and mannose in the molar ratio of 2.5 : 2.1 : 2.5. The minor components in this fraction were xylose and fucose. While the major sugar constituents of the mycelial acidic fraction of MWS-DA-GI were galactose, fucose, mannose and glucose. The trace components in this fraction was xylose. From the results of periodate oxidation, Smith degradation, affinity chromatography and methylation analysis, the chemical structures of the two fractions, BWS-DA-GI and MWS-DA-GI were both determined as ${\beta}-1,3$ glucans. It was also estimated that BWS-DA-GI had a $1{\rightarrow}6$ glucosidic linkage and MWS DA-GI had $1{\rightarrow}4$ and $1{\rightarrow}6$ glucosidic linkages. The molecular weights of these fractions, MWS-DA-GI and MWSDA-GI were estimated as $1.2{\times}10^6\;and\;1.0{\times}10^6$ dalton, respectively.

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Analysis of a Large-scale Protein Structural Interactome: Ageing Protein structures and the most important protein domain

  • Bolser, Dan;Dafas, Panos;Harrington, Richard;Schroeder, Michael;Park, Jong
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2003.10a
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    • pp.26-51
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    • 2003
  • Large scale protein interaction maps provide a new, global perspective with which to analyse protein function. PSIMAP, the Protein Structural Interactome Map, is a database of all the structurally observed interactions between superfamilies of protein domains with known three-dimensional structure in thePDB. PSIMAP incorporates both functional and evolutionary information into a single network. It makes it possible to age protein domains in terms of taxonomic diversity, interaction and function. One consequence of it is to predict the most important protein domain structure in evolution. We present a global analysis of PSIMAP using several distinct network measures relating to centrality, interactivity, fault-tolerance, and taxonomic diversity. We found the following results: ${\bullet}$ Centrality: we show that the center and barycenter of PSIMAP do not coincide, and that the superfamilies forming the barycenter relate to very general functions, while those constituting the center relate to enzymatic activity. ${\bullet}$ Interactivity: we identify the P-loop and immunoglobulin superfamilies as the most highly interactive. We successfully use connectivity and cluster index, which characterise the connectivity of a superfamily's neighbourhood, to discover superfamilies of complex I and II. This is particularly significant as the structure of complex I is not yet solved. ${\bullet}$ Taxonomic diversity: we found that highly interactive superfamilies are in general taxonomically very diverse and are thus amongst the oldest. This led to the prediction of the oldest and most important protein domain in evolution of lift. ${\bullet}$ Fault-tolerance: we found that the network is very robust as for the majority of superfamilies removal from the network will not break up the network. Overall, we can single out the P-loop containing nucleotide triphosphate hydrolases superfamily as it is the most highly connected and has the highest taxonomic diversity. In addition, this superfamily has the highest interaction rank, is the barycenter of the network (it has the shortest average path to every other superfamily in the network), and is an articulation vertex, whose removal will disconnect the network. More generally, we conclude that the graph-theoretic and taxonomic analysis of PSIMAP is an important step towards the understanding of protein function and could be an important tool for tracing the evolution of life at the molecular level.

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Survey of genetic structure of geese using novel microsatellite markers

  • Lai, Fang-Yu;Tu, Po-An;Ding, Shih-Torng;Lin, Min-Jung;Chang, Shen-Chang;Lin, En-Chung;Lo, Ling-Ling;Wang, Pei-Hwa
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.2
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    • pp.167-179
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    • 2018
  • Objective: The aim of this study was to create a set of microsatellite markers with high polymorphism for the genetic monitoring and genetic structure analysis of local goose populations. Methods: Novel microsatellite markers were isolated from the genomic DNA of white Roman geese using short tandem repeated probes. The DNA segments, including short tandem repeats, were tested for their variability among four populations of geese from the Changhua Animal Propagation Station (CAPS). The selected microsatellite markers could then be used to monitor genetic variability and study the genetic structures of geese from local geese farms. Results: 14 novel microsatellite loci were isolated. In addition to seven known loci, two multiplex sets were constructed for the detection of genetic variations in geese populations. The average of allele number, the effective number of alleles, the observed heterozygosity, the expected heterozygosity, and the polymorphism information content were 11.09, 5.145, 0.499, 0.745, and 0.705, respectively. The results of analysis of molecular variance and principal component analysis indicated a contracting white Roman cluster and a spreading Chinese cluster. In white Roman populations, the CAPS populations were depleted to roughly two clusters when K was set equal to 6 in the Bayesian cluster analysis. The founders of private farm populations had a similar genetic structure. Among the Chinese geese populations, the CAPS populations and private populations represented different clads of the phylogenetic tree and individuals from the private populations had uneven genetic characteristics according to various analyses. Conclusion: Based on this study's analyses, we suggest that the CAPS should institute a proper breeding strategy for white Roman geese to avoid further clustering. In addition, for preservation and stable quality, the Chinese geese in the CAPS and the aforementioned proper breeding scheme should be introduced to geese breeders.

Synthesis and Characteristics of Highly Oil-absorptive Expanded Polyurethane (고흡유성 발포 폴리우레탄의 합성과 특성)

  • Lee, Yong-Hun;Kim, Wook;Kim, Won-Ho
    • Polymer(Korea)
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    • v.24 no.2
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    • pp.149-158
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    • 2000
  • Oil-absorptive expanded polyurethane (EPU) was prepared with a lypophilic polyol, polypropyleneglycol (PPG) as the soft segment, and toluenediisocyanate (TDI) and $H_2O$ as the hard segment. PPGs haying various average molecular weights ((equation omitted) : 1000, 2000, 3000) were employed to investigate that the soft segment content was consequent on the oil-absorptivity and the mechanical properties of the EPUs. As (equation omitted) of PPG was decreased from 3000 to 1000, the oil-absorptivity and the tensile strength of the EPUs increased from 1460 to 3010% and from 0.26 to 0.55 $kg_{f}$ /$cm^2$ respectively. As the hard segment content ratio, ${\gamma}$ ([NCO]/[OH]) was increased from 1.0 to 1.2, the tensile strength of the EPUs increased from 0.56 to 0.95 $kg_{f}$ /$cm^2$, due to the formation of allophanate and/or biuret bondings. However, as the surfactant (S-A) content was increased from 1.0 to 2.5 pbw, the oil-absorptivity was decreased from 3634 to 3312%, due to the formation of closed cell structures.

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Assessment of Microbial Community in Paddy Soils Cultivated with Bt and Nakdong Rice (Bt 벼의 토양미생물상 영향 비교평가)

  • Sohn, Soo-In;Ahn, Byung-Ohg;Chi, Hee-Youn;Cho, Byung-Kwan;Cho, Min-Seok;Shin, Kong Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.5
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    • pp.829-835
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    • 2012
  • The cultivation of genetically modified (GM) crops has increased due to their economic and agronomic advantages. Before commercialization of GM crops, however, we must assess the potential risks of GM crops on human health and environment. The aim of this study was to investigate the possible impact of Bt rice on the soil microbial community. Microbial communities were isolated from the rhizosphere soil cultivated with Bt rice and Nakdong, parental cultivar and were subjected to be analyzed using both culture-dependent and molecular methods. The total counts of bacteria, fungi, and actinomycetes in the rhizosphere of transgenic and conventional rice were not significantly different. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that the bacterial community structures during cultural periods were very similar each other. Analysis of dominant isolates in the rhizosphere cultivated with Bt and Nakdong rice showed that the dominant isolates from the soil of Bt rice and Nakdong belonged to the Proteobacteria, Cloroflexi, Actinobacteria, Firmicutes, and Acidobacteria. These results indicate that the Bt rice has no significant impact on the soil microbial communities during cultivation period. Further study remains to be investigated whether the residue of Bt rice effect on the soil environment.

The Isolation and Pyrolysis of the Brown Pigmented Macromolecule from the Cured Leaf Tobacco (잎담배 성분중 갈색고분자 물질의 분리정제 및 열분해에 관한 연구)

  • Chae, Quae;Park, Ji-Chang
    • Journal of the Korean Society of Tobacco Science
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    • v.2 no.1
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    • pp.1-7
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    • 1980
  • Gel filtration column chromatography (Sephadex G-75), dialysis an d Brushite column chromatography were carried out to separate the brown pigmented macromolecule from water extracts of the cured leaf tobaccos. The two distinct macromolecules having different molecular weight were separated by the Sephadex column chromatography. Brushite also separated two different species of macromolecules which might have different electronic structures. According to the enzymatic degradation of protein in Burley and Hicks, chymotrypsin showed the best degradation ratio, ie., 16-30% in Burley and 38-57% in Hicks. Similar effect was observed with pepsin. However, very low effect of degradation was revealed with trypsin. The sample treated with the proteolytic enzymes revealed the disappearance of the first peak and the slight decrease of the 2nd peak height in the separation profile of Sephadex. After dialysis, the brown pigmented macromolecule was pyrolyzed at $300^{\circ}C$ and the strongly fluorescent components not identified before pyrolysis were detected with TLC separation. Absorption spectrum of these fluorescent compounds was monitored in benzene and the absorption maxima at 265nm and 275 nm were obtained. Considering absorption maxima and shape of the spectrum, those fluorescent compounds seem to be PAH derivatives.

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Media Characteristics of PVA-derivative Hydrogels Using a CGA Technique (CGA 제조기법을 응용한 PVA 하이드로젤의 담체 특성)

  • Yoon, Mi-Hae; Kwon, Sung-Hyun;Cho, Dae-Chul
    • Journal of Environmental Science International
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    • v.18 no.3
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    • pp.299-308
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    • 2009
  • We manufactured PVA-derived hydrogels using a foam generation technique that has been widely used to prepare colloidal gas aphrons(CGA). These gels were differentiated to the conventional gels such as for medical or pharmaceutical applications, which have tiny pores and some crystalline structure. Rather these should be used in de-pollution devices or adhesion of cells or biomolecules. The crosslinkers used in this work were amino acid, organic acid, sugars and lipids(vitamins). The structures of the gels were observed in a scanned electron microscope. Amino acids gels showed remarkably higher swelling ratios probably because their typical functional groups help constructing a highly crosslinked network along with hydrogen bonds. Boric acid and starch would catalyze dehydration while structuring to result in much lower water content and accordingly high gel content, leading to less elastic, hard gels. Bulky materials such as ascorbic acid or starch produced, in general, large pores in the matrices and also nicotinamide, having large hydrophobic patches was likely to enlarge pore size of its gels as well since the hydrophobicity would expel water molecules, thus leading to reduced swelling. Hydrophilicity(or hydrophobicity), functional groups which are involved in the reaction or physical linkage, and bulkiness of crosslinkers were found to be more critical to gel's cross linking structure and its density than molecular weights that seemed to be closely related to pore sizes. Microscopic observation revealed that pores were more or less homogeneous and their average sizes were $20{\mu}m$ for methionine, $10-15{\mu}m$ for citric acid, $50-70{\mu}m$ for L-ascorbic acid, $30-40{\mu}m$ for nicotinamide, and $70-80{\mu}m$ for starch. Also a sensory test showed that amino acid and glucose gels were more elastic meanwhile acid and nicotinamide gels turned out to be brittle or non-elastic at their high concentrations. The elasticity of a gel was reasonably correlated with its water content or swelling ratio. In addition, the PVA gel including 20% ascorbic acid showed fair ability of cell adherence as 0.257mg/g-hydrogel and completely degraded phenanthrene(10 mM) in 240 h.

Pyrrole-Derivative of Chalcone, (E)-3-Phenyl-1-(2-Pyrrolyl)-2-Propenone, Inhibits Inflammatory Responses via Inhibition of Src, Syk, and TAK1 Kinase Activities

  • Yang, Sungjae;Kim, Yong;Jeong, Deok;Kim, Jun Ho;Kim, Sunggyu;Son, Young-Jin;Yoo, Byong Chul;Jeong, Eun Jeong;Kim, Tae Woong;Han Lee, In-Sook;Cho, Jae Youl
    • Biomolecules & Therapeutics
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    • v.24 no.6
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    • pp.595-603
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    • 2016
  • (E)-3-Phenyl-1-(2-pyrrolyl)-2-propenone (PPP) is a pyrrole derivative of chalcone, in which the B-ring of chalcone linked to ${\beta}$-carbon is replaced by pyrrole group. While pyrrole has been studied for possible Src inhibition activity, chalcone, especially the substituents on the B-ring, has shown pharmaceutical, anti-inflammatory, and anti-oxidant properties via inhibition of NF-${\kappa}B$ activity. Our study is aimed to investigate whether this novel synthetic compound retains or enhances the pharmaceutically beneficial activities from the both structures. For this purpose, inflammatory responses of lipopolysaccharide (LPS)-treated RAW264.7 cells were analyzed. Nitric oxide (NO) production, inducible NO synthase (iNOS) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) mRNA expression, and the intracellular inflammatory signaling cascade were measured. Interestingly, PPP strongly inhibited NO release in a dose-dependent manner. To further investigate this anti-inflammatory activity, we identified molecular pathways by immunoblot analyses of nuclear fractions and whole cell lysates prepared from LPS-stimulated RAW264.7 cells with or without PPP pretreatment. The nuclear levels of p50, c-Jun, and c-Fos were significantly inhibited when cells were exposed to PPP. Moreover, according to the luciferase reporter gene assay after cotransfection with either TRIF or MyD88 in HEK293 cells, NF-${\kappa}B$-mediated luciferase activity dose-dependently diminished. Additionally, it was confirmed that PPP dampens the upstream signaling cascade of NF-${\kappa}B$ and AP-1 activation. Thus, PPP inhibited Syk, Src, and TAK1 activities induced by LPS or induced by overexpression of these genes. Therefore, our results suggest that PPP displays anti-inflammatory activity via inhibition of Syk, Src, and TAK1 activity, which may be developed as a novel anti-inflammatory drug.

Characterization of Caveola-Vesicle Complexes (CVCs) Protein, PHIST/CVC-8195 in Plasmodium vivax

  • Wang, Bo;Lu, Feng;Han, Jin-Hee;Lee, Seong-Kyun;Cheng, Yang;Nyunt, Myat Htut;Ha, Kwon-Soo;Hong, Seok-Ho;Park, Won Sun;Han, Eun-Taek
    • Parasites, Hosts and Diseases
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    • v.54 no.6
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    • pp.725-732
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    • 2016
  • Plasmodium vivax produces numerous caveola-vesicle complex (CVC) structures beneath the membrane of infected erythrocytes. Recently, a member helical interspersed subtelomeric (PHIST) superfamily protein, $PcyPHIST/CVC-81_{95}$, was identified as CVCs-associated protein in Plasmodium cynomolgi and essential for survival of this parasite. Very little information has been documented to date about $PHIST/CVC-81_{95}$ protein in P. vivax. In this study, the recombinant $PvPHIST/CVC-81_{95}$ N and C termini were expressed, and immunoreactivity was assessed using confirmed vivax malaria patients sera by protein microarray. The subcellular localization of $PvPHIST/CVC-81_{95}$ N and C termini in blood stage parasites was also determined. The antigenicity of recombinant $PvPHIST/CVC-81_{95}$ N and C terminal proteins were analyzed by using serum samples from the Republic of Korea. The results showed that immunoreactivities to these proteins had 61% and 43% sensitivity and 96.9% and 93.8% specificity, respectively. The N terminal of $PvPHIST/CVC-81_{95}$ which contains transmembrane domain and export motif (PEXEL; RxLxE/Q/D) produced CVCs location throughout the erythrocytic-stage parasites. However, no fluorescence was detected with antibodies against C terminal fragment of $PvPHIST/CVC-81_{95}$. These results suggest that the $PvPHIST/CVC-81_{95}$ is localized on the CVCs and may be immunogenic in natural infection of P. vivax.

Seroprevalence of Norwalk-Like Viruses (NLVs) in Seoul: Detection Using Baculovirus-Expressed Norwalk Virus Capsid Antigen (Baculovirus에서 발현된 Norwalk Virus 캡시드 항원에 대한 국내인의 혈청학적 반응도)

  • Kang, Shin-Jai;Nam, Ki-Bum;Cho, Yang-Ja;Kim, Kyung-Hee
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.161-168
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    • 1997
  • The present study was designed to estimate the seroprevalence of NLVs among diarrheagenic children and in healthy adults in Seoul and its vicinity with the use of an EIA and an Western blot (WB) based on recombinant Norwalk virus capsid protein (rNV) and crude virus preparations as antigen. Seroconversion was observed in 34 (83%) of 41 tested using the EIA and in 21 (54%) of 39 using the WB, suggesting that the NLVs with epitopes common to rNV are prevalent in Seoul area. Diarrheal children who were known to have been infected with several other strains of the NLVs showed no significant antibody response to the rNV. Infection with rNV occurred earlier in life: primary infections with rNV were common before the age of 6 months and over 91 % of children had evidence of infection by that age by the EIA. Since the amount of the NLV antigens available for seroepidemiologic surveys is limited, we tried to detect NLV antibody by using crude virus preparations as antigen. One crude virus preparation of a child whose stool yielded genetically distinct NLV revealed the presence of the plural number of bands upon SDS-PAGE, but precipitated only one band (62 kDa) after the WB with a serum (collected 10 days after the onset of symptoms) of another diarrheal child. The WB assay we present in this report revealed that the NLVs are prevalent among Korean population and that the sera contained antibody to a single major structural protein, with molecular sizes of 58 to 62 kDa, compatible with the sizes reported for the Norwalk virus and Snow Mountain agent proteins, respectively. When the results of the WB were compared with those obtained by the EIA, the EIA antibody assay was sensitive enough to detect an antibody rise of as much as 4096-fold but not as specific as the WB. The WB assay presented in this paper will provide a powerful tool to elucidate not only antigenic structures of the NL Vs but also seroepidemiology of the NLV infection. The availability of an unlimited source of antigen will enable a large scale serologic studies that will greatly increase our understanding of the role of NLVs in human enteric illness.

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