• Title/Summary/Keyword: Da No.

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Function of Nitric Oxide in Activation-Induced Cell Death of T Lymphocytes

  • Park, Yuk-Pheel;Paik, Sang-Gi;Kim, Young-Sang
    • Animal cells and systems
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    • v.4 no.4
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    • pp.381-388
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    • 2000
  • Using a murine T cell hybridoma, activation-induced cell death (AICD) was studied. As an in vitro model system for the AICD, 1 cell hybridoma expressing TCR/CD3 complex was incubated onto the immobilized purified anti-CD3 antibody. The immobilized anti-CD3 antibody induced AICD effectively up to 40%. At 1-100 $\mu$M range of SNP, an exogenous source of nitric oxide (NO), the cell proliferation was not affected, but at 1 mM SNP, cell proliferation was significantly reduced. The AICD of T cell hybridoma was inhibited by exogenous NO at non-cytotoxic concentration, In the cells undergoing AICD, the expressions of caspase-3 and FasL were detected, but not iNOS. Similar result was recognized in the apoptosis induced by dexamethasone, an apoptosis-inducing agent. However, the conversion from the inactive form of caspase-3 (32 kDa) to the active form (17 kDa) was significantly reduced in the cells in AICD induced by anti-CD3 antibody, With the result of increased PARP cleavage in the cells, we propose that another PARP cleavage pathway not involving caspase-3 may function in the anti-CD3 antibody induced AICD in the T cell hybridoma.

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Quantification of nitric oxide concentration in the hemocytes of Manila clam Ruditapes philippinarum by using 4,5-diaminofluorescein diacetate (DAF-2) detection method (바지락 (Ruditapes philippinarum) 혈구의 일산화질소 (nitric oxide) 정량)

  • Nam, Ki-Woong;Yang, Hyun-Sung;Park, Kyung-Il
    • The Korean Journal of Malacology
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    • v.29 no.1
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    • pp.15-21
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    • 2013
  • Nitric oxide (NO) is an important intra-intercellular signaling molecule that regulates many physiological processes and participates in the development some pathological conditions in animals. In this study, we compared different methods for determining NO concentration in the hemocytes of Manila clam Ruditapes philippinarum. For measuring the intracellular NO levels, we used the specific fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA), and the quantification methods that were compared were based on image analysis, spectrophotometry, and flow cytometry. NO concentration could be determined using all the 3 methods, and the concentration varied significantly depending upon the presence of NO regulators in the hemocytes; NO concentration increased in the presence of L-arginine, while it decreased in the presence of N-nitro-L-arginine methyl ester. In particular, it is found that estimation of NO using a flowcytometry is more economical, reliable and accurate compared to image analysis and spectrophotometry. Accordingly we believe that determining NO concentration by using flowcytometry will be useful in evaluating physiological and pathological conditions in marine bivalves.

Purification and Characterization of Antibacterial Peptides, Spodopsin Ia and Ib Induced in the Larval Haemolymph of the Common Cutworm, Spodoptera Iitura

  • Choi, Chung-Sik;Yoe, Sung-Moon;Kim, Eung-Seok;Chae, Kwon-Seok;Kim, Hak R.
    • Animal cells and systems
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    • v.1 no.3
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    • pp.457-462
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    • 1997
  • Antibacterial activity was induced in the haemolymph of the common cutworm, Spodoptera litura by the artificial injection of E, coli Ek132. Antibacterial peptides were purified from the immunized haemolymph by heat treatment, ion-exchange chromatography, gel filtration chromatography, and reverse phase FPLC, and their physicochemical characteristics were investigated. These purified antibacterial peptides designated as spodopsin la and Ib were named after Spodoptera litura. Spodopsin la and Ib had the apparent molecular masses of 3, 823 Da and 3, 886 Da, respectively, and about 20% of the sequences had basic amino acids, such as lysine and arginine but no cysteine. Also, spodopsin l was confirmed to be a new member of cecropin family having a similar amino acid sequence to cecropin of lepidopteran insects, such as Bombyx mori and Hyalophora cecropia. The purified spodopsin was active against gram-positive as well as gram-negative bacteria.

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Purification and characterization of a thermostable glutamate dehydrogenase from a thermophilic bacterium isolated from a sterilization drying oven

  • Amenabar, Maximiliano J.;Blamey, Jenny M.
    • BMB Reports
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    • v.45 no.2
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    • pp.91-95
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    • 2012
  • Glutamate dehydrogenase from axenic bacterial cultures of a new microorganism, called GWE1, isolated from the interior of a sterilization drying oven, was purified by anion-exchange and molecular-exclusion liquid chromatography. The apparent molecular mass of the native enzyme was 250.5 kDa and was shown to be an hexamer with similar subunits of molecular mass 40.5 kDa. For glutamate oxidation, the enzyme showed an optimal pH and temperature of 8.0 and $70^{\circ}C$, respectively. In contrast to other glutamate dehydrogenases isolated from bacteria, the enzyme isolated in this study can use both $NAD^+$ and $NADP^+$ as electron acceptors, displaying more affinity for $NADP^+$ than for $NAD^+$. No activity was detected with NADH or NADPH, 2-oxoglutarate and ammonia. The enzyme was exceptionally thermostable, maintaining more than 70% of activity after incubating at $100^{\circ}C$ for more than five hours suggesting being one of the most thermoestable enzymes reported in the family of dehydrogenases.

NMR Characterization of Oxidized Form of Human 8-kDa Dynein Light Chain

  • Shin, Jae-Sun;Jeong, Woo-Jin;Chi, Seung-Wook
    • Journal of the Korean Magnetic Resonance Society
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    • v.14 no.2
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    • pp.127-133
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    • 2010
  • Redox-dependent conformational change of human 8-kDa Dynein light chain (LC8) plays important role in regulating NF-${\kappa}B$ signaling pathway. In this study we characterized the structural states of the oxidized and reduced forms of LC8 by using NMR spectroscopy. The $^1H-^{15}N$ 2D HSQC spectra of oxidized LC8 indicated that no significant change in tertiary structure of LC8 occurred upon oxidation. The chemical shift perturbations of LC8 upon oxidation suggest a redox-dependent quaternary structural change.

Electron Microscopy of the Separated Outer Tegument of the Sparganum and Its Antigenicity

  • Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.50 no.2
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    • pp.181-183
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    • 2012
  • The author reported previously on separation of the outer tegument of the spargana (plerocercoids of Spirometra mansoni) using high concentration of urea solution. To determine which layer of the tegument is separated by this method, an electron microscopic analysis has been processed in this study. It was confirmed that the basement layer of the tegument is separated from the parenchyme of the sparganum. In addition, the antigenicity of the separated outer tegument against the human sparganosis patient sera was evaluated. Numerous antigenic proteins, including 16 and 55 kDa proteins, were noticed in the separated tegument; however, there were no diagnostic 31/36 kDa molecules in this tegument. The molecules reactive with the patient sera in the tegument are to be characterized in future studies.

젖소 초유 Insulin-like Growth Factor-I 분획이 in vivo에서 Murine Macrophage의 면역 활성에 미치는 영향

  • Hwang, Gyeong-A;Kim, Seung-Il;Yang, Hui-Jin;Lee, Su-Won
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.10a
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    • pp.340-344
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    • 2004
  • 젖소 초유 중에 함유된 IGF-I rich fraction을 30kDa와 1kDa ultrafiltration(UF) membrane을 이용하여 효과적으로 분리하였으며 분획내의 IGF-I은 SDS-PAGE와 Western blot으로 확인하였다. 분획한 IGF-I rich fraction이 murine macrophage의 면역 활성에 미치는 영향을 실험한 결과 $1{\mu}g$ 투여군의 경우 IL-6는 7.3ng/ml, NO는 $10.7{\mu}M$을 생성하였고 Phagocytosis는 대조구 대비하여 65% 증진시켰다. $TNF-{\alpha}$의 분비량은 L929세포의 증식 저해율로 측정한 결과 대조구에 대비하여 30% 더 높은 저해율을 나타내었고, $H_2O_2$는 대조구에 대비하여 6% 더 높은 과산화수소를 생산하였다.

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Effect of Traditional Acupuncture on Abomasal Motility in Holstein Cow (젖소에서 일반침술이 제 4위 운동성에 미치는 영향)

  • 이주명;남치주;장광호
    • Journal of Veterinary Clinics
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    • v.15 no.1
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    • pp.151-155
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    • 1998
  • Abomasal motility was observed by needling to acupoints for 20 min. The acupoints used were guan yuan yurt da chang yurt dei yup pi yurt and hou hoi. The acupoint which showed the increase of the largest wave type was pi yu and the increase of wave type was observed 20 minutes after needling in hou hoi and dei yu acupoint. But there was no effect on abomasal motility after needling to guan yuan yu and da chang yu. On the other hands change in amplitude of the abomasal contraction after needling to these acupoints was not abserved. These results indicate that stimulation to pi yu acupoint would be the most useful to increase the abomasal motility.

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TOTAL COLORING OF MIDDLE GRAPH OF CERTAIN SNAKE GRAPH FAMILIES

  • A. PUNITHA;G. JAYARAMAN
    • Journal of applied mathematics & informatics
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    • v.42 no.2
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    • pp.353-366
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    • 2024
  • A total coloring of a graph G is an assignment of colors to both the vertices and edges of G, such that no two adjacent or incident vertices and edges of G are assigned the same colors. In this paper, we have discussed the total coloring of M(Tn), M(Dn), M(DTn), M(ATn), M(DA(Tn)), M(Qn), M(AQn) and also obtained the total chromatic number of M(Tn), M(Dn), M(DTn), M(ATn), M(DA(Tn)), M(Qn), M(AQn).

Reduction of Muscarinic $K^+$ Channel Activity by Transferrin in Ischemic Rat Atrial Myocytes

  • Park, Kyeong-Tae;Kang, Da-Won;Han, Jae-Hee;Hur, Chang-Gi;Hong, Seong-Geun
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.6
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    • pp.333-339
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    • 2003
  • It has been demonstrated that an unidentified cytosolic factor(s) reduces $K_{ACh}$ channel function. Therefore, this study attempted to elucidate the cytosolic factor. Fresh cytosol isolated from normal heart (FC) depressed the $K_{ACh}$ channel activity, but cytosol isolated from the ischemic hearts (IC) did not modulate the channel function. Electrophorectic analysis revealed that a protein of ${\sim}80 kDa was markedly reduced or even lost in IC. By using peptide sequencing analysis and Western blot, this 80 kDa protein was identified as transferrin (receptor-mediated $Fe^{3+}$ transporter, 76 kDa). Direct application of transferrin (100 nM) to the cytoplasmic side of inside-out patches decreased the open probability ($P_o$, 12.7${\pm}6.4%, n=4) without change in mean open time (${\tau}_o$, $98.5{\pm}1.3$%, n=4). However, the equimolar apotransferrin, which is free of $Fe^{3+}$, had no effect on the channel activity (N*$P_o$, $129.1{\pm}13.5$%, n=3). Directly applied $Fe^{3+}$ (100 nM) showed results similar to those of transferrin (N*$P_o$: $21.1{\pm}3.9$%, n=5). However $Fe^{2+}$ failed to reduce the channel function (N*$P_o$, $106.3{\pm}26.8$%, n=5). Interestingly, trivalent cation La3+ inhibited N*$P_o$ of the channel ($6.1{\pm}3.0$%, n=3). Taken together, these results suggest that $Fe^{3+}$ bound to transferrin can modulate the $K_{ACh}$ channel function by its electrical property as a polyvalent cation.