• Title/Summary/Keyword: M-13

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Effect of Phorbol ester on $K^+$channel in an G292 osteoblast-like cell (G292 세포에서 $K^+$통로에 대한 phorbol ester의 효과)

  • Kim, Mi-Kyung;Park, Su-Byung
    • The korean journal of orthodontics
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    • v.32 no.3 s.92
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    • pp.227-234
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    • 2002
  • In order to investigate the action mechanism of protein kinase C on $K^+$ channel in osteoblastic cell, effects of phorbol 12, 13-dibutyrate on human osteoblast-like cells (G292) were studied by patch clamp technique with cell-attacked configuration. 111 this experiment, 45pS ion channel was dominant in G292 cell line according to their approximate conductances in symmetrical 140mM KCl saline at holding potential of 60mV. In torrent-voltage relationship, reversal potential was 5.5mV at the condition of potassium enriched saline in the pipette and -27 mV at the condition of standard extracellular saline In the pipette. Phorbol 12, 13-dibutyrate 10nM increased the open probability of 45pS channel and staurosporine, an inhibitor of protein kinase C, suppressed this effect. Phorbol 12,13-dibutyrate moved the reversal potential of 45pS channel to more negative potential and increased the single channel current at the same membrame potential. In order to check the activation of protein kinase C in G292 cell by phorbol 12,13-dibutyrate, western blot of protein kinase C was performed. Phorbol 12,13-dibutyrate $0.1{\mu}M$ translocated protein kinase C from cellular compartment to membrane compartment of the cell. These findings suggest that phorbol 12,13-dibutyrate, one of phorbol esters, activate 45pS channel In G292 cell and affect cell membrane potential, that regulate cellular function.

EFFECT OF PATULIN ON THE GROWTH OF BACTERIOPHAGE M13

  • Lee, Kil-Soo
    • Toxicological Research
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    • v.5 no.1
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    • pp.53-59
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    • 1989
  • A mycotoxin Patulin, isolated from apple juice medium cultured with Penicillium patulum NRRL5259, was purified through acid aluminum column using ethyl ether as eluent. The yield of purified patulin crystal was 3mg/ml culture medium after 8 days of shaking culture at 28C. The growth rate of Escherichia coli K12JM103 infected with bacteriophage M13 was decreased by patulin at the concentration range of 1Mug/ml to 10Mu/nl. ED50 of patulin for the bacterial growth was 4.5Mug/ml and 10Mug/ml of patulin caused maximum inhibitory effect (60% inhibition) on the growth.

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Characterization of Kraft Lignin by 13C-Nuclear Magnetic Resonance Spectroscopy (13C-NMR에 의한 크라프트 리그닌의 특성)

  • Lee, Beom-Goo;Hwang, Byung-Ho
    • Journal of the Korean Wood Science and Technology
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    • v.17 no.4
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    • pp.44-56
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    • 1989
  • To recover much residual lignin from the black liquor of kraft pulp. the black liquor was extracted and purified with many organic solvents. Many kinds of lignins were isolated from each fraction obtained and the characteristics of these lignins investigated by $^{13}C$-NMR spectroscopy. If M$\ddot{o}$rck's method was compared with Kirk's method. M$\ddot{o}$rck's method was better than Kirk's method because the particular signals of each lignin occur more in the former than m the latter. Especially the $^{13}C$-NMR spectrum of the MCS fraction identifies with those of other researchers. The experiment that the kraft lignin from Pinus densiflora S. et Z. found in Korea was investigated by $^{13}C$-NMR spectroscopy was performed first in Korea.

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PCR-Based Detection of Mycoplasma Species

  • Sung Hyeran;Kang Seung Hye;Bae Yoon Jin;Hong Jin Tae;Chung Youn Bok;Lee Chong-Kil;Song Sukgil
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.42-49
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    • 2006
  • In this study, we describe our newly-developed sensitive two-stage PCR procedure for the detection of 13 common mycoplasmal contaminants (M. arthritidis, M. bovis, M. fermentans, M. genitalium, M. hominis, M. hyorhinis, M. neurolyticum, M. orale, M. pirum, M. pneumoniae, M. pulmonis, M. salivarium, U. urealyticum). For primary amplification, the DNA regions encompassing the 16S and 23S rRNA genes of 13 species were targeted using general mycoplasma primers. The primary PCR products were then subjected to secondary nested PCR, using two different primer pair sets, designed via the multiple alignment of nucleotide sequences obtained from the 13 mycoplasmal species. The nested PCR, which generated DNA fragments of 165-353 bp, was found to be able to detect 1-2 copies of the target DNA, and evidenced no cross-reactivity with the generated DNA of related microorganisms or of human cell lines, thereby confirming the sensitivity and specificity of the primers used. The identification of contaminated species was' achieved via the performance of restriction fragment length polymorphism (RFLP) coupled with Sau3AI digestion. The results obtained in this study furnish evidence suggesting that the employed assay system constitutes an effective tool for the disagnosis of mycoplasmal contamination in cell culture systems.

Measurement of Various Dimensions of the Larynx in Korea Adult (한국인 성인의 후두계측에 관한 연구)

  • 박승훈;박성남;김미자;윤희병;정대현;장혁순;전승하;강주원
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1982.05a
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    • pp.9.1-9
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    • 1982
  • The authors have measured various dimensions of the larynx of twenty adult cadavers. (male; 13, female; 7) The result were as follows, 1) Transverse diameter between the greater horns of the hyoid bone; M.52.58mm, F.43.07mm. 2) Transverse diameter between the lesser horns of the hyoid bone; M.36.20mm, F.30.20mm. 3) Transverse diameter of the thyroid cartilage; M.50.58mm, F.42.58mm. 4) Transverse diameter of the cricoid cartlage; M.30.14mm, F.26.94mm. 5) Distance from the lower margin of the hyoid bone to the thyroid notch; M.12.83mm, F.10.92mm. 6) Distance from the lower margin of the thyroid cartilage to the lower margin of the cricoid cartilage; M.16.40mm, F.10.26mm. 7) Distance from the upper margin of the thyroid cartilage to the lower margin of the cricoid cartilage; M.34.65mm, F.34.61mm. 8) Anteroposterior diameter of the cricoid cartilage; M.25.32mm, F.20.01mm. 9) Transverse diameter of the epiglottis; M.29.32mm, F.22.08mm. 10) Vertical diameter of the epiglottis; M.35.90mm, F.27.90mm. 11) Transverse diameter between the cuneiform tubercles; M.17.69mm, F.13.52mm. 12) Length of the vocal fold; M.11.61mm, F.10.40mm. 13) Transvers diameter of the vocal fold; M.18.62mm, F.18.27mm. 14) Distance from the upper margin of the epiglottis to the vocal fold; M.42.45mm, F.34.52mm. 15) Anteroposterior length of the vocal fold; M.19.25mm, F.10.70mm.

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Purification and Properties of .$\beta$-1, 3-Glucanase from Pseudomonas stutzeri KF13 (Pseudomonas-stutzeri KF13의 ..$\beta$-1, 3-Glucanase 정제 및 성질)

  • 방광웅;송형익;김재근;유대식;정기택
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.1-8
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    • 1987
  • An extracellular $\beta$-1, 3-glucanase from Pseudomonas stutzeri KF 13 was purified about 390 with 26% recovery. The purified enzyme revealed a single band by polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis. The enzyme was stable in a pH 6.0 to 9.0, and relatively thermostable. The optimal pH and temperature on the enzyme activity were found to be 5.8 and 45.deg.C, respectively. The activation energy was calculated to be 16,130 cal per mole. The Km value for laminarin was found to be 3ng per ml and the molecular weight was determined to be 28,000 by gel filtration and 26,000 daltons by SDS-acrylamide gel electrophoresis. The enzyme was inhibited by 1.0mM of $Hg^{2+}$, and strongly inhibited by 1.0mM of p-chloromercuribenzoic acid.

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Prolyl Endopeptidase-inhibiting Isoflavonoids from Puerariae Flos and Some Revision of their $^{13}C-NMR$ Assignment (갈화의 Prolyl Endopeptidase 저해 활성 Isoflavonoid 및 이들의 $^{13}C-NMR$ Assignment)

  • Kim, Kyung-Bum;Kim, Sang-In;Kim, Jong-Sik;Song, Kyung-Sik
    • Applied Biological Chemistry
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    • v.42 no.4
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    • pp.351-355
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    • 1999
  • In order to find anti-dementia drugs from natural products, prolyl endopeptidase inhibitors were purified from Puerariae Flos by consecutive solvent partition, followed by silica gel, Sephadex LH-20, and HPLC. Four isoflavonoid inhibitors were isolated and identified as tectorigenin, genistein, 5,7-dihydroxy-4',6-dimethoxyisoflavone, and 5-hydroxy-6,7,4'-trimethoxyisoflavone by means of instrumental analyses including $^{1}H-$, $^{13}C-$, $^{2}D-NMR$ and MS and $IC_{50}$ values against PEP were 5.30 ppm$(17.7\;{\mu}M)$, 10.39 ppm$(38.5\;{\mu}M)$, 13.92 ppm$(44.3\;{\mu}M)$, and 20.61 ppm$(62.8\;{\mu}M)$, respectively. Some previous mistakes in $^{13}C-NMR$ assignment were revised by careful investigation of HMBC and HMQC data.

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Inhibitory Effect of Phorbol 12-Myristate 13-Acetate on NO Production Induced by Interleukin-1 beta in Aortic Vascular Smooth Muscle Cells of Rats (혈관평활근세포에서 Phorbol 12-Myristate 13-Acetate의 전처리가 Interleukin-1β에 의한 Nitrite생성에 미치는 영향)

  • 윤병헌;김인겸;박태규;김중영
    • Journal of Life Science
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    • v.13 no.4
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    • pp.441-447
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    • 2003
  • To examine the role of protein kinase C (PKC) in regulation of interleukin-1 beta (IL-1$\beta$)-induced iNOS expression, IL-1$\beta$-induced nitrite production was observed in cultured vascular smooth muscle (VSM) cells pretreated with phorbol 12-myristate 13-acetate (PMA) and phorbol 12,13-butyrate (PDB) as PKC activator; 4$\alpha$-phorbol-didecanoate (PDD) as PKC non-activator. Nitrite production induced by IL-1$\beta$ was increased by the presence of increasing concentration of PMA ranging from 2 to 200 nM. However, in VSM cells pretreated with PMA and PDB, IL-1$\beta$-induced $NO_2$ production was decreased in proportion to the duration of pretreatment, and most significantly decreased in pretreatment time of 24 hours. Using RT-PCR method, the expression of iNOS mRNA induced by IL-1$\beta$ was decreased in VSM cells pretreated with PMA 200 nM for 24 hours. These results suggest that decrease in IL-I$\beta$-induced nitrite production by the pretreatment of PMA result from inhibition of iNOS expression and the inhibition related to PMA-induced PKC down-regulation.

mPW1PW91 Calculated Relative Stabilities and Structures for the Conformers of 1,3-dimethoxy-p-tert-butylthiacalix[4]crown-5-ether (1,3-디메톡시-티아캘릭스[4]크라운-5-에테르의 이형체들의 상대적인 안정성과 구조들에 대한 mPW1PW91 계산 연구)

  • Kim, Kwang-ho;Choe, Jong-In
    • Journal of the Korean Chemical Society
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    • v.53 no.5
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    • pp.521-529
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    • 2009
  • Molecular structures of the various conformers for the 1,3-dimethoxy-p-tert-butylthiacalix[4] crown-5-ether (3) were optimized by using DFT B3LYP/6 - 31 + G(d,p) and mPW1PW91/6 - 31 + G(d,p) (hybrid HF-DF) calculation methods. We have analyzed the energy differences and structures of eight in/out orientations (cone_oo, cone_oi, pc_oo, pc_io, pc_oi, pc_ii, 13a_oo, 13a_io) of two methoxy groups in three major conformations (cone, partial-cone and 1,3-alternate). The 13a_oo (out-out orientation of the 1,3-alternate conformer) is calculated to be the most stable among eight different conformations of 3, and in accord with the experimental result. The ordering of relative stability resulted from the mPW1PW91/6 - 31 + G(d,p) calculation method is following: 13a_oo > 13a_io$\sim$pc_io$\sim$cone_oo > cone_oi$\sim$pc_oo$\sim$pc_oi > pc_ii.

Removal of Nitrogens and Phosphorus by Bacillus sp. CK-11 and Bacillus sp. CK-13 Isolated from Shrimp Farming Pond (새우양식장에서 분리한 Bacillus sp. CK-10과 Bacillus sp. CK-13에 의한 질소와 인의 제거)

  • Chun Jae-Woo;Ma Chae-Woo;Lee Sang-Hyun;Oh Kye-Heon
    • KSBB Journal
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    • v.20 no.2 s.91
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    • pp.116-122
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    • 2005
  • The feasibility of using bacterial cultures with the ultimate aim for the marine environmental clean-up was explored. The present study reports on the bacterial elimination of nitrogens and phosphorus by strains CK-10 and CK-13 isolated from shrimp farming pond. The strains were identified as genus Bacillus on the basis of BIOLOG test, and designated as Bacillus sp. CK-10 and Bacillus sp. CK-13, respectively. Removal of nitrogens $(NH_4^+,\;NO_2^-,\;or\;NO_3^-)$ or phosphorus $(PO_4^{-3})$ as single N or P source was studied with single cultures under aerobic conditions. Complete elimination of all nitrogens in the concentration range of $100-400{\mu}M$ was achieved in single cultures as well as co-cultures within the given incubation period. Similar results were obtained from the test cultures containing $125-599{\mu}M\;PO_4^{3-}$. Simultaneous removal of all N/P was monitored in the co-cultures. As the results, $400{\mu}M\;NH_4^+\;and\;NO_2^-$ were eliminated within 12hrs and $400{\mu}M\;NO_3^-\;and\;500{\mu}M\;PO_4^{-3}$ were completely disappeared within 36 hrs from the media. The work demonstrated that co-cultures improved the concurrent removal of N/P from the media.