• Title/Summary/Keyword: NMR study

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Characterization and Preparation of Low Molecular Weight Water Soluble Chitosan Nanoparticle Modified with Cell Targeting Ligand for Efficient Gene Delivery (효과적인 유전자전달을 위한 표적성 리간드가 도입된 저분자량 수용성 키토산 나노입자의 제조 및 특성)

  • Heo, Sun-Heang;Jang, Min-Ja;Kim, Dong-Gon;Jeong, Young-Il;Jang, Mi-Kyeong;Nah, Jae-Woon
    • Polymer(Korea)
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    • v.31 no.5
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    • pp.454-459
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    • 2007
  • Gene therapy using low molecular weight water soluble chitosan (LMWSC) as polycationic polymer shows good biocompatibility, but low transfection efficiency. The mechanism of folic acid (FA) uptake in the cells to promote targeting and internalization could improve transfection rates. The objective of this study was to synthesize and characterize the WSCFA-DNA complex and evaluate their cytotoxicity, in vitro. In $^1H-NMR$ spectra, specific peaks appeared both of FA and LMWSC in $D_2O$. WSCFA nanoparticles have spherical shapes with particle size show below 110 nm. In the cell cytotoxicity test, the WSCFA-DNA complex showed high cell viability, in vitro. Gel electrophoresis showed condensed DNA within the carriers. hi vitro transfection efficiency was assayed by fluorescence spectroscopy WSCFA nanoparticles have less cytotoxicity, good DNA condensation and particle size around 110 nm, which makes them a promising candidate as a non-viral gene vector.

Studies on toxic substances of Gingko leaves inducing contact dermatitis in rabbit (은행엽(銀杏葉)의 가토(家兎) 피부(皮膚) 접촉성염증(接觸性炎症) 유발(誘發) 독성물질(毒成物質)에 관(關)한 연구(硏究))

  • Baek, Byeong-kirl;Kim, Byeong-su;Kim, Sun-hee;Ahn, Byung-zun
    • Korean Journal of Veterinary Research
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    • v.31 no.3
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    • pp.319-327
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    • 1991
  • Gingko biloba is one of the oldest trees on earth. It can be traced 300 million years or more into past. In recent years, this tree has become increasingly popular because of its longevity. Amount of comsumption of Gingko leaves in Korea is increased to produce drugs for blood circulating system's disorder. The workers who work at the company of product drug with Gingko leaves complain pruritus at belt position of west, however toxic substance from extracts of Gingko biloba was carried out to determine what kind of the toxic substance from extract could induce a contact dermatitis by means of histopathological study. Dried leaves of Korean G. biloba were exhaustively extracted with chloroform. The aqueous chloroform residue was extracted with hexane. After evaporation, the hexane extract was passed through a silica gel column chromatography, it was separated into 9 fractions by means of gradient solvent (benzene: acetone/ 98~93 : 2~7%). The 4th and 7th fraction were idendified as bilibol and gingkolic acid by silica gel thin layer chromatography and NMR spectrophotometer. The 4th and 7th fractions of Korean Gingko leaves have an irritant substances which can be induced ear swelling and increased thickness of stratum cornium of rabbit ear exposed with the fractions (3 time/3 day, $100{\mu}l$ of 50mg/ml ethyl alcohol). In the 4th fraction, the substance have induced the surface of ear to swell and mass of unknown cell like inflammatory cell were infiltrated within the stratum corneum, and the 7th fraction have the surface of ear to loss the hairs, and sebaceous gland and follicle of hair were infiltrated with inflammatory cells. and other fractions at all did not induced a certain contact dermatitis and also Deutschen drug of Gingko (named as Gerigeron) have induced the contact dermatitis such as dematitis of Korean Gingko extract.

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Inhibitory Effect of Flavan-3-ols Isolated from Oolong Tea on Xanthine Oxidase (우롱차로부터 분리된 Flavan-3-ol 화합물의 Xanthine Oxidase에 대한 저해 영향)

  • An, Bong-Jeun;Bae, Man-Jong;Choi, Chung
    • Korean Journal of Food Science and Technology
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    • v.28 no.6
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    • pp.1084-1088
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    • 1996
  • As a series of study for an anti-gout agent, two flavan-3-ols have been isolated from Oolong tea. They showed positive reaction in $anisaldehyde-H_2SO_4$ solution and $FeCl_3$, which were able to be confirmed in TLC with dimeric flavan-3-ols. The components were identified as procyanidin B-2 and procyanidin B-2-O-3, 3'-di-gallate by spectroscopic analysis. Their inhibitory effect on xanthine oxidase was also investigated and procyanidin B-2-O-3, 3'-digallate showed 60.6% inhibition at $50\;{\mu}mole$. The compound competitively inhibited the xantine oxidase and dimeric flavan-3-ols contanining gallate had higher inhibition activity.

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Identification of 3'-Hydroxymelanetin and Liquiritigenin as Akt Protein Kinase Inhibitors

  • Yang Hye-Young;Lee Hong-Sub;Ko Jong-Hee;Yeon Seung-Woo;Kim Tae-Yong;Hwang Bang-Yeon;Kang Sang-Sun;Chun Jae-Sun;Hong Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1384-1391
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    • 2006
  • The signal transduction system is one of the most important devices involved in maintaining life, and many protein kinases are included in the cellular signal transduction system. Finding a protein kinase inhibitor is very valuable, as it can be used to study cell biology and applied to pharmaceuticals. For the efficient and rapid screening of protein kinase inhibitors, two assay systems were combined; the nonradioactive protein kinase assay system that uses an FITC-labeled IRS-2 peptide and the cell-based paper disc assay system that uses Streptomyces griseus as the indicator strain. Among 330 kinds of herb extracts tested, the extract of Dalbergia odorifera exhibited the strongest inhibitory activity in the two assay systems and was selected for further isolation. Based on solvent extraction and many steps of chromatography, seven compounds were finally separated to homogeneity and their structures determined by $^{1}H$ and $^{13}C$ NMR spectroscopies. Four were to be flavonoids and identified as butin ($C_{15}H_{12}O_5$, Mw=272.07), 3'-hydroxymelanetin ($C_{16}H_{12}O_6$, Mw=300.06), liquiritigenin ($C_{15}H_{12}O_4$, Mw=256.07), and 2'-hydroxyformononetin ($C_{16}H_{12}O_{5}$, Mw=284.07). 3'-Hydroxymelanetin inhibited the phosphorylation of the GSK3 protein by Akt to 37% at a concentration of $10{\mu}g/ml$ and showed the strongest cytotoxicity ($ED_{50}<50{\mu}g/ml$) against the human cancer cell line HCT116. Under the same conditions, liquiritigenin also inhibited the phosphorylation of GSK3 by Akt to 26%, and its cytotoxicity against the HCT116 cell line was lower than $100{\mu}g/ml$.

Ovicidal Activity of Lactic Acid Produced by Lysobacter capsici YS1215 on Eggs of Root-Knot Nematode, Meloidogyne incognita

  • Lee, Yong Seong;Naning, Kyaw Wai;Nguyen, Xuan Hoa;Kim, Sun Bae;Moon, Jae Hak;Kim, Kil Yong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.11
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    • pp.1510-1515
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    • 2014
  • Lysobacter capsici YS1215 isolated from soil previously showed nematicidal potential for biological control of the root-knot nematode. In this study, lactic acid, a nematicidal compound, was isolated from culture filtrate of YS1215, and its ovicidal activity was investigated. Purification and identification of lactic acid were performed by a series of column chromatographies and identified by $^1H$ and $^{13}C$ NMR spectra and GC-MS analysis. Our results showed that bacterial culture filtrate containing lactic acid significantly inhibited egg hatching. The lowest egg hatch rate (5.9%) was found at a high concentration ($25 {\mu}l/ml$) of lactic acid at 5 days after incubation, followed by 20 (15.2%), 15 (23.7%), 10 (29.8%), and $5(36.4%){\mu}l/ml$, while egg hatching in the control (sterile distilled water) was 44.5%. This is the first report of lactic acid as an ovicidal compound, and it may be considered as an alternative of chemical pesticide against root-knot nematodes.

Biruloquinone, an Acetylcholinesterase Inhibitor Produced by Lichen-Forming Fungus Cladonia macilenta

  • Luo, Heng;Li, Changtian;Kim, Jin Cheol;Liu, Yanpeng;Jung, Jae Sung;Koh, Young Jin;Hur, Jae-Seoun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.161-166
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    • 2013
  • At present, acetylcholinesterase (AChE) inhibitors are the first group of drugs to treat mild to moderate Alzheimer's disease (AD). Although beneficial in improving cognitive and behavioral symptoms, the effectiveness of AChE inhibitors has been questioned since they do not delay or prevent neurodegeneration in AD patients. Therefore, in the present study, in order to develop new and effective anti-AD agents from lichen products, both the AChE inhibitory and the neuroprotective effects were evaluated. The AChE inhibitory assay was performed based on Ellman's reaction, and the neuroprotective effect was evaluated by using the MTT method on injured PC12 cells. One AChE inhibitor ($IC_{50}$ = 27.1 ${\mu}g/ml$) was isolated by means of bioactivity-guided isolation from the extract of lichen-forming fungus Cladonia macilenta, which showed the most potent AChE inhibitory activity in previous screening experiment. It was then identified as biruloquinone by MS, and $^1H$- and $^{13}C$-NMR analyses. The inhibitory kinetic assay suggested that biruloquinone is a mixed-II inhibitor on AChE. Meanwhile, biruloquinone improved the viability of the $H_2O_2$- and ${\beta}$-amyloid-injured PC12 cells at 1 to 25 ${\mu}g/ml$. The protective effects are proposed to be related to the potent antioxidant activities of biruloquinone. These results imply that biruloquinone has the potential to be developed as a multifunctional anti- AD agent.

Characterization and Antioxidant Activity of the Exopolysaccharide Produced by Bacillus amyloliquefaciens GSBa-1

  • Zhao, Wen;Zhang, Jian;Jiang, Yun-Yun;Zhao, Xiao;Hao, Xiao-Na;Li, Liu;Yang, Zhen-Nai
    • Journal of Microbiology and Biotechnology
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    • v.28 no.8
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    • pp.1282-1292
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    • 2018
  • The exopolysaccharide (EPS) produced by Bacillus amyloliquefaciens GSBa-1 was isolated and purified by ethanol precipitation, and DEAE-cellulose and Sepharose CL-6B chromatographies. The molecular mass of the purified EPS was determined to be 54 kDa. Monosaccharide analysis showed that the EPS was composed of predominantly glucose, and it was further confirmed by NMR spectroscopy to be ${\alpha}-glucan$ that consisted of a trisaccharide repeating unit with possible presence of two ${\alpha}-(1{\rightarrow}3)$ and one ${\alpha}-(1{\rightarrow}6)$ glucosidic linkages. Microstructural analysis showed that the EPS appeared as ellipsoid or globose with a smooth surface. The EPS had a degradation temperature at $240^{\circ}C$. Furthermore, the EPS had strong DPPH and hydroxyl radical scavenging activities, and moderate superoxidant anion scavenging and metal ion-chelating activities. This is the first characterization of a glucan produced by B. amyloliquefaciens with strong antioxidant activity. The results of this study suggest the potential of the EPS from B. amyloliquefaciens GSBa-1 to serve as a natural antioxidant for application in functional products.

Potentiometric Study of Transition Metal Ions Complexes of Hexadentate($N_4$, $O_2$) Ligands (질소-산소계 여섯 자리 리간드들의 전이금속이온 착물에 대한 전위차법 연구)

  • Kim, Sun-Deuk;Kim, Jun-Kwang;Lee, Woo-Sik
    • Journal of the Korean Chemical Society
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    • v.44 no.6
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    • pp.518-525
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    • 2000
  • The open-chain hexadentate N$_4$, O$_2$ ligands 1,13-bis(2-hydroxybenzyl)-2,5,9,12-tetraazatridecane-tetrahydrochloride (BSATD${\cdot}$4HCl) and 1,14-bis (2-hydroxybenzyl)-2,6,9,12-tetraazatetradecane-tetrahydrochloride (BSATED${\cdot}$ 4HCl) have been synthesized as their tetrahydrochloride salt and characterized by EA, IR, NMR and Mass. Their proton dissociation constants(log$K^{n}_{H}$) and stability constants(log$K_{ML}$) for Cu(II), Ni(II), Co(II), and Zn(II) ions were determined in aqueous solution by potentiometry and compared with those of analogous N$_4$, O$_2$ ligands contain ethylenic spacers or propylenic spacers, which make six-membered chelate rings between the aliphatic nitrogen atoms. Synthesis and characterization of [Cu(BSATD)]ClO$_4$ and [Cu(BSATED)](ClO$_4$)$_2$ complexes are described.

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A Study on the Photo-Degradation Properties of the Spiropyran Using THz-TDS (테라헤르츠 시간 영역 분광법을 이용한 스피로파이란의 광 퇴화 특성 연구)

  • Bang, Jin-Hyuk;Park, Myoung-Hwan;Ryu, Han-Cheol
    • Journal of the Korean Chemical Society
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    • v.60 no.1
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    • pp.28-33
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    • 2016
  • The spiropyran is a typical material having photodegradation properties in the process of photochromism. The spiropyran has been utilized in various applications such as optical switch, optical memories, and biosensor because of its remarkable stability, fast responsive time, stronger color change, and photo-induced controllability. However, the spriropyran is photodegraded by the repetitive optical irradiation. The photodegradation of spiropyran have been investigated by using UV-Visible spectroscopy, nuclear magnetic resonance (NMR), and Raman spectroscopy. Herein, the properties of spiropyran were characterized by using terahertz time-domain spectroscopy (THz-TDS) in the terahertz frequency region. In terahertz region, the measured absorbance of spiropyran was increased due to the photodegradation induced by the repetitive UV irradiation. The absorbance tendency of spiropyran in the terahertz frequency region was compared with that in the visible region, and they were completely opposite to each other.

Structural and Functional Characterization of CRAMP-18 Derived from a Cathelicidin-Related Antimicrobial Peptide CRAMP

  • Park, Kyong-Soo;Shin, Song-Yub;Hahm, Kyung-Soo;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.24 no.10
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    • pp.1478-1484
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    • 2003
  • CRAMP was identified from a cDNA clone derived from a mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptide. Tertiary structure of CRAMP in TFE/$H_2O$ (1 : 1, v/v) solution has been determined by NMR spectroscopy previously and consists of two amphipathic $\alpha-helices$ from Leu4 to Lys10 and from Gly16 to Leu33. These two helices are connected by a flexible region from Gly11 to Gly16. Analysis of series of fragments composed of various portion of CRAMP revealed that an 18-residue fragment with the sequence from Gly16 to Leu33 (CRAMP-18) was found to retain antibacterial activity without cytotoxicity. The effects of two Phe residues at positions 14 and 15 of CRAMP-18 on structure, antibacterial activity, and interaction with lipid membranes were investigated by $Phe^{14,15}$ ${\rightarrow}$ Ala substitution (CRAMP-18-A) in the present study. Substitution of Phe with Ala in CRAMP-18 caused a significant reduction on antibacterial and membrane-disrupting activities. Tertiary structures of CRAMP-18 in 50% TFE/$H_2O$ (1 : 1, v : v) solution shows amphipathic ${\alpha}$-helix, from $Glu^2{\;}to{\;}Leu^{18}$, while CRAMP-18-A has relatively short amphipathic ${\alpha}$-helix from $Leu^4{\;}to{\;}Ala^{15}$. These results suggest that the hydrophobic property of $Phe^{14}{\;}and{\;}Phe^15$ in CRAMP-18 is essential for its antibacterial activity, ${\alpha}$-helical structure, and interactions with phospholipid membranes.