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Effect of $K^+-channel$ Blockers on the Muscarinic- and $A_1-adenosine-Receptor$ Coupled Regulation of Electrically Evoked Acetylcholine Release in the Rat Hippocampus

  • Yu, Byung-Sik;Kim, Do-Kyung;Choi, Bong-Kyu
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.2
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    • pp.147-154
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    • 1998
  • It was attempted to clarify the participation of $K^+-channels$ in the post-receptor mechanisms of the muscarinic and $A_1-adenosine$ receptor- mediated control of acetylcholine (ACh) release in the present study. Slices from the rat hippocampus were equilibrated with $[^3H]$choline and the release of the labelled products was evoked by electrical stimulation (3 Hz, 5 V/cm, 2 ms, rectangular pulses), and the influence of various agents on the evoked tritium-outflow was investigated. Oxotremorine (Oxo, $0.1{\sim}10\;{\mu}M$), a muscarinic agonist, and $N^6-cyclopentyladenosine$ (CPA, $1{\sim}30\;{\mu}M$), a specific $A_1-adenosine$ agonist, decreased the ACh release in a dose-dependent manner, without affecting the basal rate of release. 4-aminopyridine (4AP), a specific A-type $K^+-channel$ blocker ($1{\sim}100\;{\mu}M$), increased the evoked ACh release in a dose-related fashion, and the basal rate of release is increased by 3 and $100\;{\mu}M$. Tetraethylammonium (TEA), a non-specific $K^+-channel$ blocker ($0.1{\sim}10\;{\mu}M$), increased the evoked ACh release in a dose-dependent manner without affecting the basal release. The effects of Oxo and CPA were not affected by $3\;{\mu}M$ 4AP co-treatment, but 10 mM TEA significantly inhibited the effects of Oxo and CPA. 4AP ($10\;{\mu}M$)- and TEA (10 mM)-induced increments of evoked ACh release were completely abolished in Ca^{2+}-free$ medium, but these were recoverd in low Ca^{2+}$ medium. And the effects of $K^+-channel$ blockers in low Ca^{2+}$ medium were inhibited by $Mg^{2+}$ (4 mM) and abolished by $0.3\;{\mu}M$ tetrodotoxin (TTX). These results suggest that the changes in TEA-sensitive potassium channel permeability and the consequent limitation of Ca^{2+}$ influx are partly involved in the presynaptic modulation of the evoked ACh-release by muscarinic and $A_1-adenosine$ receptors of the rat hippocampus.

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The Influence of Surface Treatments on Shear Bond Strength between Zirconia Core and Heat Press Ceramic Interface (지르코니아 코어의 표면처리방법이 열 가압 세라믹과의 전단결합강도에 미치는 영향)

  • Park, Hang-Min;Chung, In-Sung
    • Journal of Technologic Dentistry
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    • v.31 no.2
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    • pp.23-30
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    • 2009
  • All-ceramic restorations have gained acceptance among clinicians and patients because of their superior esthetics. Most all-ceramic systems have a 2-layer structure, using a weak veneering ceramic over a strong supporting core. often, failure of all-ceramic restorations occurs when the veneering ceramic fractures, exposing the core material. The purpose of this study was to compare the shear bond strength of heat press ceramic system (Zirpress) to zirconia core with various surface treatments. 10 metal cores and 50 zirconia cores were fabricated and divided into six groups according to surface treatment such as Zirliner application, aluminium oxide blasting, and 9.5% HF etching. Sixty specimens were prepared using Zirpress, veneered 8mm height and 3mm in diameter, over the zirconia cores (n=10). The shear bond strength test was performed in a universal testing machine with a crosshead speed of 1/min. Ultimate shear bond strength data were analyzed with One-way ANOVA and the Scheffe's test (p=.05). Within the limits of this study, the following conclusions were drawn: The mean shear bond strengths (MPa) were: 12.93 for $110{\mu}m$ aluminium oxide blasting/Rexillium III/IPS e.Max Zirpress; 14.92 for $50{\mu}m$ aluminium oxide blasting ${\pm}9.5%$ HF etching/Zirconis core/IPS e.Max Zirpress; 16.37 for $110{\mu}$ aluminium oxide blasting + 9.5% HF etching/Zirconis core/IPS e.Max Zirpress; 12.89 for $200{\mu}$ aluminium oxide blasting + 9.5% HF etching/Zirconis core/IPS e.Max Zirpress; 19.30 for 9.5% HF etching/Zirconis core/IPS e.Max Zirpress; 19.55 for Zirliner/Zirconis core/IPS e.Max Zirpress. The mean shear bond strength for ZNTZH (Zirliner/Zirconis core) and ZNTEH (9.5% HF etching/Zirconis core) were significantly superior to MS110H ($110{\mu}$ aluminium oxide blasting/Rexillium III) and ZS200EH ($200{\mu}$ aluminium oxide blasting + 9.5% HF etching/Zirconis core) (p<0.05).

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Effects of Ammonium, and Nitrate on Callus Growth of Tobacco and Soybean and Activities of Nitrogen Metabolizing Enzymes (암모늄태와 질산태 질소가 담배와 콩의 조직배양시 생육 및 질소대사 효소의 활성에 미치는 영향)

  • 박혜선;설종호;장매희
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.57-61
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    • 1998
  • Soybean and tobacco tissue cultured with modified MS media containing 4 different ratio(as N) of nitrate to ammonium combination which were 3:0, 2:1, 1:2 and 0:3. The highest callus growth in soybean were observed in the 2:1 medium. The medium containing nitrate only was detrimental to soybean callus growth. Tobacco callus grown with nitrate-only grew as well as those in the 2:1 and very slowly with ammonium-only. In tobacco callus, the total nitrogen in the callus increased with the increase of nitrogen concentration in the medium, but in soybean callus, the opposite result was noted. Nitrate reductase activity in tobacco callus was high when grown with nitrate-only but low with ammonium-only. In case of soybean callus, nitrate reductase activity was high in the 2:1 and remarkably low in nitrate-only medium. Both in soybean and tobacco callus, the activity of glutamine synthetase was high with nitrate-only, but low with ammonium.

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Chemical Constituents of Domestic Quercus spp. Leaves (국내산 참나무속 수종 잎의 추출성분)

  • Kim, Jin-Kyu;Bae, Young-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.6
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    • pp.61-71
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    • 2006
  • This study was carried out to investigate chemotaxonomical correlation an d the chemical constituents of domestic Quercus sp. leaves. The leaves of Q. mongolica, Q. aliena, Q. serrata, Q. acutissima, Q. dentata and Q. variabilis were collected in the experimental forest of Kangwon National University. The combined extracts were successively fractionated with n -hexane, methylene chloride and ethyl acetate using a separation funnel. A portion of the ethyl acetate and $H_2O$ soluble materials of each species were chromatographed on a Sephadex LH-20 column using various aqueous MeOH and EtOH-hexane as washing solvents. Spectrometric analysis such as NMR and MS, including TLC, were performed to characterize the structures of the isolated compounds. Gallic acid, (+)-catechin, (-)-epicatechin, (+)-gallocatechin, kaempferol, astragalin, astragalin-6"-O-gallate, isoquercitrin, isoquercitrin-6"-O-gallate and myricetin were isolated from Q. mongolic a leaves. Gallic acid, kaempferol and quercetin were characterized from Q. acutissima leaves. Gallic acid, (+)-catechin, (-)-epicatechin, (+)-gallocatechin, (-)-epigallocatechin, kaempferol, quercetin, guajaverin and tamarixin were identified from Q. dentata leaves. Gallic acid, (+)-catechin, (-)-epicatechin, kaempferol, quercitrin, isoquercitrin and myricetin were purified from Q. serrata leaves. Gallic acid, (+)-catechin, astragalin, astragalin-6"-O-gallate and isoquercitrin were isolated from Q. variabilis leaves. Gallic acid was isolated from all the leaves and could be a taxonomic index on Quercus spp..

Blood Aluminum Concentrations among Residents of the South-east Costal Area of Korea (우리나라 일부 지역 주민들의 혈중 알루미늄 농도)

  • Seo, Jeong-Wook;Kim, Byoung-Gwon;Kim, Yu-mi;Chung, Jin-Yong;Lim, Hyoun-Ju;Ye, Byeong-Jin;Hong, Young-Seoub
    • Journal of Environmental Health Sciences
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    • v.42 no.3
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    • pp.196-204
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    • 2016
  • Objectives: Aluminum is well known as a potent neurotoxicant. There are many reports that aluminum can be toxic to humans and to animals. However, there are only few studies on the assessment of aluminum exposure among humans in Korea. Therefore, the aim of this study was to evaluate current aluminum concentrations among the adult population in regions of Korea. Methods: We selected 439 adults aged 20-89 years from the Busan-Ulsan-Gyeongnam region and certain other sites. Blood aluminum concentrations were analyzed using inductively-coupled plasma mass spectrometry (ICP/MS). The geometric means (GM) and 95% confidence intervals (CI) of aluminum concentrations were calculated, and we also confirmed the proportion of excess of the aluminum reference value. Results: Total GM (95% CI) of aluminum levels was 14.26 (13.43-15.14) ppb. Levels among males (15.58 [14.09-17.22] ppb) were higher compared to among females (13.51 [12.54-14.55] ppb), and levels increased with subject age from 40 years and over. Three point one nine percent of the subjects exceed the reference value of 50 ppb. Lastly, aluminum concentration has a log-normal distribution with lnN (x; 2.89, 0.642), x>0. Conclusion: In the present study, we evaluated the distribution of blood aluminum concentrations among the normal population in Korea, and we found some adults exceeding the reference levels. However, in order to compare the results with other studies, an extended study including measurement of serum aluminum level is required. In addition, further research on various population groups, including occupational exposure of workers, is required.

Development of HPLC Determination Method for Trace Levels of 1-, 2-Nitropyrenes and 2-Nitrofluoranthene in Airborne Particulates and Its Application to Samples Collected at Noto Peninsula

  • Hayakawa, Kazuichi;Tang, Ning;Sato, Kosuke;Izaki, Akihiko;Tatematsu, Michiya;Hama, Hirotaka;Li, Ying;Kameda, Takayuki;Toriba, Akira
    • Asian Journal of Atmospheric Environment
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    • v.5 no.3
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    • pp.146-151
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    • 2011
  • 1-Nitropyrene (1-NP), 2-NP and 2-nitrofluoranthene (2-NFR) are useful markers for studying the atmospheric behaviors of polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (NPAHs). However, present methods for measuring trace levels of these compounds are lesssensitive and laborious. Here we describe several improvements to a previously reported high-performance liquid chromatography-chemiluminescence detection system that allows it to determine trace levels of 1-, 2-NPs and 2-NFR. The proposed system was equipped with a reducer column packed with Pt/Rh instead of zinc whose life-time was limited. The combination of Cosmosil MS-II (monomeric ODS) and AR-II (polymeric ODS) columns was used instead of polymeric ODS columns as the separator column to improve the separation. An ethanol mixture with acetate buffer (pH 5.5) was used in place of an acetonitrile mixture with the same buffer to activate the reducer column. The same ethanol mixture was used as the mobile phase for the clean-up column. The switching time of the column switching valve was optimized to concentrate the amino-derivatives of above NPAHs quantitatively on the concentrator column. The concentrations of bis(2,4,6-trichlorophenly) oxalate and hydrogen peroxide in the chemiluminescence reagent solution were optimized to 0.4 mM and 30 mM, respectively, to increase the sensitivity. Under the above conditions, the detection limits (S/N=3) of 1-, 2-NPs and 2-NFR were 1 fmol (0.25 pg), 10 fmol (2.5 pg) and 4 fmol (1 pg), respectively. The proposed system was effectively used to determine trace levels of 1-, 2-NPs and 2-NFR in airborne particulates collected at Noto Peninsula. The atmospheric concentrations of 1-, 2-NPs and 2-NFR were not more than sub pg $m^{-3}$ levels. They were higher in winter (January) than in summer (July). In both seasons, the concentrations were in decreasing order, [2-NFR]>[1-NP]>[2-NP].

Callus Induction and Plant Regeneration from Mature Seeds of Timothy (티모시 성숙종자로부터 캘러스 유도 및 식물체 재분화)

  • Lee, Ki-Won;Kim, Ki-Yong;Choi, Gi-Jun;Lim, Young-Chul;Kim, Won-Ho;Jung, Min-Wong;Seo, Sung;Lee, Byung-Hyun;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.28 no.3
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    • pp.165-170
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    • 2008
  • Timothy (Phleum pratense L.) is an important grass species as forage. In order to optimize tissue culture conditions of timothy, the effects of plant growth regulators on callus induction and plant regeneration was investigated with mature seeds of colt cultivar. The optimal concentration of 2,4-D for the induction of primary callus from mature seeds was 3 mg/L. The highest embryogenic callus frequenc (25%) was observed when the mature seed were cultured on MS medium supplemented with 3 mg/L 2,4-D and 0.1 mg/L BA. The highest plant regeneration frequency was observed when type B callus was transferred to N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Regenerated plants were grown normally when shoots were transplanted to the soil. A short tissue culture period and regeneration system would be beneficial for molecular breeding of timothy by the production of transgenic plant.

Heterologous Expression and Characterization of Glycogen Branching Enzyme from Synechocystis sp. PCC6803

  • Lee, Byung-Hoo;Yoo, Young-Hee;Ryu, Je-Hoon;Kim, Tae-Jip;Yoo, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1386-1392
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    • 2008
  • A gene (sll0158) putatively encoding a glycogen branching enzyme (GBE, E.C. 2.4.1.18) was cloned from Synechocystis sp. PCC6803, and the recombinant protein expressed and characterized. The PCR-amplified putative GBE gene was ligated into a pET-21a plasmid vector harboring a T7 promoter, and the recombinant DNA transformed into a host cell, E. coli BL21(DE3). The IPTG-induced enzymes were then extracted and purified using Ni-NTA affinity chromatography. The putative GBE gene was found to be composed of 2,310 nucleotides and encoded 770 amino acids, corresponding to approx. 90.7 kDa, as confirmed by SDS-PAGE and MALDI-TOF-MS analyses. The optimal conditions for GBE activity were investigated by measuring the absorbance change in iodine affinity, and shown to be pH 8.0 and $30^{\circ}C$ in a 50 mM glycine-NaOH buffer. The action pattern of the GBE on amylose, an $\alpha$-(1,4)-linked linear glucan, was analyzed using high-performance anion-exchange chromatography (HPAEC) after isoamylolysis. As a result, the GBE displayed $\alpha$-glucosyl transferring activity by cleaving the $\alpha$-(1,4)-linkages and transferring the cleaved maltoglycosyl moiety to form new $\alpha$-(1,6)-branch linkages. A time-course study of the GBE reaction was carried out with biosynthetic amylose (BSAM; $M_p{\cong}$8,000), and the changes in the branch-chain length distribution were evaluated. When increasing the reaction time up to 48 h, the weight- and number-average DP ($DP_w$ and $DP_n$) decreased from 19.6 to 8.7 and from 17.6 to 7.8, respectively. The molecular size ($M_p$, peak $M_w{\cong}2.45-2.75{\times}10^5$) of the GBE-reacted product from BSAM reached the size of amylose (AM) in botanical starch, yet the product was highly soluble and stable in water, unlike AM molecules. Thus, GBE-generated products can provide new food and non-food applications, owing to their unique physical properties.

Inhibition of Low Density Lipoprotein-oxidation, ACAT-1, and ACAT-2 by Lignans from the Bark of Machilus thunbergii

  • Shrestha, Sabina;Park, Ji-Hae;Lee, Dae-Young;Cho, Jin-Gyeong;Lee, Do-Gyeong;Cho, Moon-Hee;Jeong, Tae-Sook;Kang, Hee-Cheol;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.63-66
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    • 2011
  • The bark of Machilus thunbergii was extracted with 80% aqueous methanol (MeOH), and the concentrated extract was partitioned using ethyl acetate (EtOAc), butanol (n-BuOH), and $H_2O$, successively. From the EtOAc fraction, five lignans were isolated through the repeated silica gel, octadecyl silica gel (ODS) and, Sephadex LH-20 column chromatography. Based on nuclear magnetic resonance (NMR), mass spectroscopy (MS), and infrared spectroscopy (IR) spectroscopic data, the chemical structures of the compounds were determined to be machilin A (1), machilin F (2), licarin A (3), nectandrin A (4), and nectandrin B, (5). This study presents comparative account of five lignans from M. thunbergii bark contributing inhibition of low density lipoprotein (LDL), ACAT-1, and ACAT-2. Compounds 2-5 showed varied degree of antioxidant activity on LDL with $IC_{50}$ values of 2.1, 11.8, 15.3, and $4.1{\mu}M$. Compounds 1, 2, and 3 showed inhibition activity on ACAT-1 with values $63.4{\pm}6.9%$ ($IC_{50}=66.8{\mu}M$), $53.7{\pm}0.9%$ ($IC_{50}=109.2{\mu}M$), and $78.7{\pm}0.2%$ ($IC_{50}=40.6{\mu}M$), respectively, at a concentration of 50 mg/mL, and on ACAT-2 with values $47.3{\pm}1.5%$ ($IC_{50}=149.7{\mu}M$), $39.2{\pm}0.2%$ ($IC_{50}=165.2{\mu}M$), and $52.1{\pm}1.0%$ ($IC_{50}=131.0{\mu}M$, respectively, at a concentration of 50 mg/mL.

Establishment of Analytical Method for Pencycuron in Representative Agricultural Commodities by High-Performance Liquid Chromatography (대표 농산물 중 살균제 Pencycuron의 HPLC 정밀 잔류분석법 개발)

  • Lee, Hyeri;Choi, Hoon;Kim, Byung-Joon;Kim, Eunhye;Kim, Su-Hee;Lee, Jin-Beom;Lee, Young Deuk;Kim, Jeong-Han
    • The Korean Journal of Pesticide Science
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    • v.21 no.1
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    • pp.75-83
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    • 2017
  • The single residue analytical method was developed for determining fungicide pencycuron residues in various agricultural commodities with high-performance liquid chromatography (HPLC). Pencycuron residue was extracted with acetone from representative crops such as Korean cabbage, apple, brown rice and green pepper. After ethyl acetate/n-hexane partition and subsequent clean-up with silica gel chromatography, pencycuron residue was quantified by reversed phase HPLC with UV detection at 240 nm. The suspected residue of pencycuron was confirmed using selected-ion monitoring (SIM) LC/mass spectrometry (MS). Instrumental limit of quantitation (ILOQ) and method LOQ (MLOQ) were set at 2 ng and 0.02 mg/kg, respectively. Overall recoveries of pencycuron from different crop samples fortified at three levels (MLOQ, 10MLOQ, 100MLOQ) were 72~108%. This proposed method could be useful as official analytical method for quantification of pencycuron residues in agricultural commodities.