• Title/Summary/Keyword: $^1H-^{13}C\

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Detection of Mitotic Centromere-Associated Kinesin (MCAK) During Cell-Cycle Progression of Human Jurkat T Cells Using Polyclonal Antibody Raised Against Its N- Terminal Region Overexpressed in E. coli

  • Jun, Do-Youn;Rue, Seok-Woo;Kim, Byung-Woo;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.912-918
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    • 2003
  • Mitotic centromere-associated kinesin (MCAK), which is a novel kinesin with a central motor domain, is believed to playa role in mitotic segregation of chromosome during the M phase of the cell cycle. In the present study, it is shown that a rabbit polyclonal antibody has been produced using the N-terminal region (187 aa) of human MCAK expressed in E. coli as the antigen. To express the N-terminal region in E. coli, the MCAK cDNA fragment encoding N-terminal 187 aa was obtained by PCR and was then inserted into the pET 3d expression vector. Molecular mass of the N-terminal region overexpressed in the presence of IPTG was 23.2 kDa on SDS-PAGE, and the protein was insoluble and mainly localized in the inclusion body that could be easily purified from the other cellular proteins. The N-terminal region was purified by electro-elution from the gel after the inclusion body was resolved on the SDS-PAGE. The antiserum obtained after tertiary immunization with the purified protein specifically recognized HsMCAK when subjected to Western blot analysis, and showed a fluctuation of the protein level during the cell cycle of human Jurkat T cells. Synchronization of the cell-cycle progression required for recovery of cells at a specific stage of the cell cycle was performed by either hydroxyurea or nocadazole, and subsequent release from each blocking at 2, 4, and 7 h. Northern and Western analyses revealed that both mRNA and protein of HsMCAK reached a maximum level in the S phase and declined to a basal level in the G1 phase. These results indicate that a polyclonal antibody raised against the N-terminal region (187 aa) of HsMCAK, overexpressed in E. coli, specifically detects HsMCAK (81 kDa), and it can analyze the differential expression of HsMCAK protein during the cell cycle.

A Study on Salt Intake and Urinary Sodium Excretion by Groups Educated Regarding Low Salt Diet (일부 저염식 피교육자의 식염섭취 및 뇨중 Sodium 배설양상 -장류 및 김치류 등 고식염 함유 식품을 중심으로-)

  • Kim, K.S.;Shin, D.C.;Lee, S.J.;Kim, H.K.
    • Journal of Nutrition and Health
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    • v.13 no.4
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    • pp.187-194
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    • 1980
  • To estimate the amount of daily salt intake by source of food and urinary sodium excretion, a dietary survey was conducted from August 27th to September 3, 1980. The salt concentration of six kinds of food was measured. The families of 25 womens' club leaders and 25 families of hypertensive patients were studied after they had been given a short course of education on lowsalt diet. The results were as follows: 1. The average amount of high-salt foods eaten daily by the study group were as follows: 4.5 Gm. in the form of table salt 16.8 ml of home-made soy sauce 6.20 ml of commercial soy sauce 11.9 Gm. of home-made red-pepper bean paste 12.0 Gm. of home-made soy-bean paste 120.7 Gm. of Kimchi 2. The average NaCl concentration of each food was as follows: 99.5% in table salt, 22.5% in home made soy sauce, 11.9% in Commercial soy sauce, 6.8 Gm% in home made red pepper bean paste, 9.2 Gm% in home-made soy-bean paste and the average concentration of various kinds of Kimchi was 2.7 Gm%. 3. The total amount of daily salt intake per adult was calculated as 14.3 Gm. By sources of food 4.5 Gm from table salt, 3.9 Gm from home-made soy sauce and 3. 3 Gm from Kimchi were taken daily Three other kinds of food were also minor sources of salt intake.4. The average amounts of individual daily salt intake were not significantly different between the hypertensive group and the normotensive group: the median of the normotensive group (11.7Gm.) was significantly lower than that of the hypertensive group (14.9Gm.). Therefore the womens' club leaders appeared to respond more quickly than the hypertensive group after low-salt diet education. 5. The average amount of sodium excretion in the 24 hour urine specimen was 234.7 mEq. From this finding the daily NaCl intake was estimated to be 15.8Gm.

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Detection of Clavibacter michiganensis subsp. michiganensis Assisted by Micro-Raman Spectroscopy under Laboratory Conditions

  • Perez, Moises Roberto Vallejo;Contreras, Hugo Ricardo Navarro;Herrera, Jesus A. Sosa;Avila, Jose Pablo Lara;Tobias, Hugo Magdaleno Ramirez;Martinez, Fernando Diaz-Barriga;Ramirez, Rogelio Flores;Vazquez, Angel Gabriel Rodriguez
    • The Plant Pathology Journal
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    • v.34 no.5
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    • pp.381-392
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    • 2018
  • Clavibacter michiganensis subsp. michiganesis (Cmm) is a quarantine-worthy pest in $M{\acute{e}}xico$. The implementation and validation of new technologies is necessary to reduce the time for bacterial detection in laboratory conditions and Raman spectroscopy is an ambitious technology that has all of the features needed to characterize and identify bacteria. Under controlled conditions a contagion process was induced with Cmm, the disease epidemiology was monitored. Micro-Raman spectroscopy ($532nm\;{\lambda}$ laser) technique was evaluated its performance at assisting on Cmm detection through its characteristic Raman spectrum fingerprint. Our experiment was conducted with tomato plants in a completely randomized block experimental design (13 plants ${\times}$ 4 rows). The Cmm infection was confirmed by 16S rDNA and plants showed symptoms from 48 to 72 h after inoculation, the evolution of the incidence and severity on plant population varied over time and it kept an aggregated spatial pattern. The contagion process reached 79% just 24 days after the epidemic was induced. Micro-Raman spectroscopy proved its speed, efficiency and usefulness as a non-destructive method for the preliminary detection of Cmm. Carotenoid specific bands with wavelengths at 1146 and $1510cm^{-1}$ were the distinguishable markers. Chemometric analyses showed the best performance by the implementation of PCA-LDA supervised classification algorithms applied over Raman spectrum data with 100% of performance in metrics of classifiers (sensitivity, specificity, accuracy, negative and positive predictive value) that allowed us to differentiate Cmm from other endophytic bacteria (Bacillus and Pantoea). The unsupervised KMeans algorithm showed good performance (100, 96, 98, 91 y 100%, respectively).

A Survey on the Demand of Education topic for the Development of Dental Hygienist Coordination System (Dental Hygienist Coordination System 개발 교육주제 요구조사)

  • Lee, Yun-Jung;Woo, Hee-Sun
    • Journal of dental hygiene science
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    • v.10 no.5
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    • pp.349-356
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    • 2010
  • The purpose of this study was to examine the awareness of dental hygienists, who played a significant role in dental sector, about health care management and their needs for education in an attempt to pave the way for the development of a Dental Hygienist Coordination System(DHCS). The subjects were 156 dental hygienists at 85 dental clinics, who were selected in Gwangju based on the data released as of August 2008 by the Dental Association and public health centers in sampling region. A Questionnaire was conducted in person to gather survey data and SPSS 12.0 program was utilized to make a statistical analysis. This results findings suggested that a systematic curriculum should be developed by focusing on coordination theory and practice, counseling and educational practice, health insurance affairs and health insurance fee claim management.

The Physicochemical Properties of Seed Mash Prepared with Koji (Koji를 이용한 seed mash의 이화학적 특성)

  • Lee, Myung-Koo;Park, Jung-Kil;Lee, Si-Kyung
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.199-205
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    • 2005
  • Physicochemical quality characteristics of seed mashes were investigated for development of new creative breadmaking in bakery. Aridity of seed mashes fermented with Koji, lactic acid bacteria, and baker's yeast was slightly lower than those fermented with Koji and yeast. ${\alpha}-Amylase$, saccharifying amylase, and acidic protease activities of seed mash composed of water (560 mL) and Koji (400 g) were 0.26 SKB, 36 SP, and 645 HUT/g, respectively. Reducing sugar content of seed mash made with Koji increased up to 13.04% after 36 hr fermentation, then decreased drastically thereafter, whereas that of seed mashes made with Koji, yeast, and lactic acid bacteria increased up to 6.5% at 6 hr, decreased to 1 to 2.5% at 12 hr, and remained less than 0.7% after 18 hr fermentation. Total organic acid contents were 10.4-12.25mg/mL. Flavor compounds including ethyl acetate, ethyl caprylate, isoamyl acetate, and p-vinyl guaiacol were detected in seed mash fermented with Koji, yeast, and L. brevis. These results show use of seed mash fermented with Koji, S. cerevisiae, and lactic acid bacteria enhances bread flavor.

DAMGO, a ${\mu}-Opioid$ Agonist and Cholecystokinin-Octapeptide Have Dual Modulatory Effects on Capsaicin-Activated Current in Rat Dorsal Root Ganglion Neurons

  • Eun, Su-Yong;Kim, Ji-Mok;Lee, Ji-Hye;Jung, Sung-Jun;Park, Joo-Min;Park, Yun-Kyung;Kim, Dong-Kwan;Kim, Sang-Jeong;Kwak, Ji-Yeon;Kim, Jun
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.1
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    • pp.71-78
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    • 2001
  • Capsaicin, a pungent ingredient of hot pepper, elicits an intense burning pain when applied cutaneously and intradermally. Activation of capsaicin-gated channel in C-type dorsal root ganglion (DRG) neurons produces nonselective cationic currents. Although electrophysiological and biochemical properties of capsaicin-activated current $(I_{CAP})$ were studied, the regulatory mechanism and intracellular signaling pathway are still unclear. In the present study, we investigated the modulations of $I_{CAP}$ by DAMGO $({\mu}-opioid\;agonist)$ and cholecystokinin octapeptide (CCK-8). In 18 out of 86 cells, the amplitude of $I_{CAP}$ was significantly increased by DAMGO and completely reversed after washout, while $I_{CAP}$ was decreased by DAMGO in 25 cells. In 43 cells, DAMGO had no effect on $I_{CAP}$. Mean action potential duration was significantly different between 'increased-by-DAMGO' group and 'decreased-by-DAMGO' group. Mean amplitudes of $I_H$ were not significantly different between both groups. CCK-8 reversibly enhanced the amplitude of $I_{CAP}$ (5/13). DAMGO also increased $I_{CAP}$ amplitude significantly in the same cells. The amplitude of $I_{CAP}$ was increased in additive manner by combined applications of DAMGO and CCK-8 in these cells. These results suggest that DAMGO and CCK-8 can either increase or decrease $I_{CAP}$ presumably depending on the subtypes of DRG cells and classified by electrophysiological properties.

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Difference in sensitivity of Eleocharis kuroguwai tubers to bensulfuron-methyl at different burial depths (Bensulfuron-methyl에 대(對한) 올방개 괴경(塊莖) 이식(移植) 심도별(深度別) 감수성(感受性) 차이(差異))

  • Shin, H.S.;Chun, J.C.
    • Korean Journal of Weed Science
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    • v.13 no.1
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    • pp.55-61
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    • 1993
  • Difference in sensitivity of Eleocharis kuroguwai tubers to bensulfuron- methyl{methyl 2-[[[[[(4, 6-dimethoxy-2-pyrimidinyl) amino] carbonyl] amino] sulfonyl] methyl] benzoate} at different burial depths were determined with respect to tuber emergence, sprouting of lateral buds, carbohydrate consumption of the tuber, and growth and new tuber production of the regrown plants. Days required to regrowth from the growth cessation due to bensulfuron-methyl were shorter in shallow-buried tubers than in deep-buried tubers. With application of bensulfuron-methyl shallow-buried tubers consumed less carbohydrate in the tuber than deep-buried tubers as compared with in deep-buried tubers during the period of growth cessation and greater regrowth also occurred in the former. Fast and great regrowth in shallow-buried tubers resulted in great production of dry matter and new tubers. However, the differences obtained were not due to bensulfuron-methyl, but due mainly to ecological emergence and growth characteristics of tubers buried at different depths.

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Antimelanogenic Effect and Whitening of Anthocyanin Rich Fraction from Seeds of Liriope platyphylla (맥문동 종실 안토시아닌 분획물의 멜라닌 생성 억제 및 미백 효과)

  • Choung, Myoung Gun;Hwang, Young Sun;Kim, Gi Ppeum;Ahn, Kyung Geun;Shim, Hoon Seob;Hong, Seung Beom;Choi, Jae Hoo;Yu, Chang Yeon;Chung, Ill Min;Kim, Seung Hyun;Lim, Jung Dae
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.5
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    • pp.361-371
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    • 2013
  • This study was performed to determine the antimelanogenic effect and tyrosinase inhibitory activities of anthocyanin rich fraction (AN-SLP) from Liriope platyphylla Wang et Tang seeds. Anthocyanins isolated from L. platyphylla seeds revealed the presence of four major anthocyanin components, which were tentatively identified as delphinidin-3-Oglucoside, delphinidin-3-O-rutinoside, petunidin-3-O-rutinoside, and malvidin-3-O-rutinoside using semipreparative HPLC, $^1H$-NMR, $^{13}C$ NMR, FAB-MS and LC/ES-MS. The inhibitory effect of AN-SLP on tyrosinase activity was studied using in vitro (against mushroom tyrosinase) and ex vivo (against B16 melanoma cell tyrosinase) models. Cellular tyrosinase activity was decreased by AN-SLP treatment in B 16 melanoma cells through dose dependent manner, but AN-SLP did not inhibit mushroom tyrosinase and L-DOPA oxidation directly. AN-SLP showed melanin inhibition by 53.2% at 50 ${\mu}g/m{\ell}$ which was 0.7 times more efficient than the antimelanogenic effect of commercial arbutin and kojic acid (36.5%) also did not show cell toxicity. Additionally, AN-SLP inhibited the activity of ${\alpha}$-glucosidase and the glycosylation of tyrosinase in melanoma cell. The resulting unsaturated glycosylation of tyrosinase makes it unstable and disturb correct transportation. From theses results, we conclude that AN-SLP could be used as anti-melanogenic agent for skin whitening.

Evaluation of Fracture Toughness in Steel Weldment for Inner Wall of LNG Storage Tank (LNG 저장탱크 내조용 강 용접부의 파괴인성 평가)

  • Jang J.-i.;Ju J.-B.;Yang Y.-c.;Kim W.-s.;Hong S. H.;Kwon D.
    • Journal of the Korean Institute of Gas
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    • v.2 no.1
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    • pp.7-13
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    • 1998
  • In this study, for the safety performance of LNG storage tank, the fracture toughness in X-grooved weld HAZ(heat-affected zone) of $9\%$ Ni steel was evaluated qualitatively and quantitatively, and the relation with the change in microstructure was analyzed. The toughness assessment was peformed through the modified CTOD test proposed for thick weldment with X-groove. Additionally, microstructures of HAZ were evaluated by OM, SEM and XRD. From the results, HAZ toughness of SMA(shielded metal arc)-welded $9\%$ Ni steel decreased as the evaluated region approached the fusion line. The decrease in toughness was apparently caused by the increase in the fraction of coarse-grained zone within HAZ. On the other hand, toughness drop with decreasing test temperature in F.L.(fusion line) ${\~}$F.L.+3mm was larger than that in F.L.+5mm${\~}$F.L.+7mm region due to the fact that in the former regions, retained austenite had poor stability.

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Understanding of a Korean Standard for the Analysis of Hexavalent Chromium in Soils and Interpretation of their Results (토양오염공정시험기준 6가크롬 분석의 이해와 결과 해석)

  • Kim, Rog-Young;Jung, Goo-Bok;Sung, Jwa-Kyung;Lee, Ju-Young;Jang, Byoung-Choon;Yun, Hong-Bae;Lee, Yee-Jin;Song, You-Seong;Kim, Won-Il;Lee, Jong-Sik;Ha, Sang-Keun
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.5
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    • pp.727-733
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    • 2011
  • A new Korean standard for the determination of Cr(VI) in soils has been officially published as ES 07408.1 in 2009. This analytical method is based on the hot alkaline digestion and colorimetric detection prescribed by U.S. EPA method 3060A and 7196A. The hot alkaline digestion accomplished using 0.28 M $Na_2CO_3$ and 0.5 M NaOH solution (pH 13.4) at $90{\sim}95^{\circ}C$ determines total Cr(VI) in soils extracting all forms of Cr(VI), including water-soluble, adsorbed, precipitated, and mineral-bound chromates. This aggressive alkaline digestion, however, proved to be problematic for certain soils which contain large amounts of soluble humic substances or active manganese oxides. Cr(III) could be oxidized to Cr(VI) by manganese oxides during the strong alkaline extraction, resulting in overestimation (positive error) of Cr(VI). In contrast, Cr(VI) reduction by dissolved humic matter or Fe(II) could occur during the neutralization and acidic colorimetric detection procedure, resulting in underestimation (negative error) of Cr(VI). Futhermore, dissolved humic matter hampered the colorimetric detection of Cr(VI) using UV/Vis spectrophotometer due to the strong coloration of the filtrate, resulting in overestimation (positive error) of Cr(VI). Without understanding the mechanisms of Cr(VI) and Cr(III) transformation during the analysis it could be difficult to operate the experiment in laboratory and to evaluate the Cr(VI) results. For this reason, in this paper we described the theoretical principles and limitations of Cr(VI) analysis and provided useful guidelines for laboratory work and Cr(VI) data analysis.