• Title/Summary/Keyword: single cell MS

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Thecharacters of Ca2+ activated Cl- channel and its role in the cardiac myocytes (심장세포에서 세포내 Ca2+ 증가에 의해 활성화되는 Cl- 통로의 특성과 역할)

  • Park, Choon-ok;Kim, Yang-mi;Haan, Jae-hee;Hong, Seong-geun
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.25-36
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    • 1994
  • The inward tail current after a short depolarizing pulse has been known as Na-Ca exchange current activated by intracellular calcium which forms late plateau of the action potential in rabbit atrial myocytes. Chloride conductance which is also dependent upon calcium concentration has been reported as a possible tail current in many other excitable tissues. Thus, in order to investigate the exsitance of the calcium activated chloride current and its contribution to tail current, whole cell voltage clamp measurement has been made in single atrial cells of the rabbit. The current was recorded during repolarization following a brief 2 ms depolarizing pulse to +40mV from a holding potential of -70mV. When voltage-sensitive transient outward current was blocked by 2 mM 4-aminopyridine or replacement potassium with cesium, the tail current were abolished by ryanodine$(1{\mu}M)$ or diltiazem$(10{\mu}M)$ and turned out to be calcium dependent. The magnitudes of the tail currents were increased when intracellular chloride concentration was increased to 131 mM from 21 mM. The current was decreased by extracellular sodium reduction when intracellular chloride concentration was low(21 mM), but it was little affected by extracellular sodium reduction when intracellual chloride concentration was high(131 mM). The current-voltage relationship of the difference current before and after extracellular sodium reduction, shows an exponential voltage dependence with the largest magnitude of the current occurring at negative potentials, with is similar to current-voltage relationship at negative potentials, which is similar to current-voltage relationship of Na-Ca exchange current. The current was also decreased by $10{\mu}M$ niflumic acid and 1 mM bumetanide, which is well known anion channel blockers. The reversal potentials shifted according to changes in chloride concentration. The current-voltage relationships of the niflumic acid-sensitive currents in high and low concentration of chloride were well fitted to those predicted as chloride current. From the above results, it is concluded that calcium activated chloride component exists in the tail current with Na-Ca exchange current and it shows the reversal of tail current. Therefore it is thought that in the physiologic condition it leads to rapid end of action potential which inhibits calcium influx and it contributes to maintain the low intracellular calcium concentration with Na-Ca exchange mechanism.

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Bond Strength of TiO2 Coatings onto FTO Glass for a Dye-sensitized Solar Cell

  • Lee, Deuk Yong;Kim, Jin-Tae;Kim, Young-Hun;Lee, In-Kyu;Lee, Myung-Hyun;Kim, Bae-Yeon
    • Journal of Sensor Science and Technology
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    • v.21 no.6
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    • pp.395-401
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    • 2012
  • The bond strength of three types of $TiO_2$ coatings onto fluorine-doped $SnO_2$ (FTO) glass was investigated with the aid of a tape test according to ASTM D 3359-95. Transmittance was then measured using an UV-vis spectrophotometer in the wavelength range of 300 nm to 800 nm to evaluate the extent of adhesion of $TiO_2$ nanorods/nanoparticles on FTO glass. A sharp interface between the coating layer and the substrate was observed for single $TiO_2$ coating ($TiO_2$ nanorods/FTO glass), which may be detrimental to the bonding strength. In multicoating sample ($TiO_2$ nanorod/$TiO_2$ nanoparticle/$TiO_2$ nanoparticle/FTO glass), the tape test was not performed due to severe peeling-off prior to the test. On the other hand, the dual coating sample ($TiO_2$ nanorod/$TiO_2$ nanoparticle/FTO glass) showed minimum variation of transmittance (4%) after the test, suggesting that the topcoat adheres well with the FTO substrate due to the presence of the $TiO_2$ nanoparticle buffer layer. The use of a $TiO_2$ nanorod electrode layer with good adhesion may be attributed to the excellent dye sensitized solar cell performance.

Purification and Characterization of Paclitaxel from Plant Cell Cultures of Taxus chinensis in Large-Scale Process (식물세포 Taxus chinensis 배양으로부터의 Paclitaxel 대량 정제 및 특성)

  • 김진현;기은숙;민범찬;최형균;홍승서;이현수
    • KSBB Journal
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    • v.15 no.5
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    • pp.537-540
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    • 2000
  • In developing a HPLC purification process, it was hoped that a single chromatographic system would be sufficient to abtain pure paclitaxel in high yield. However, no such system was found, due in part to the complex taxoid profile of crude paclitaxel and to the rigorous nature of the product specification. A two step HPLC purification was adopted using reverse-phase separation on C(sub)18 as a first step, and normal-phase separation on silica as the final polishing step. Impurity profiles were established and maintained for paclitaxel, which identified and quantified each impurity observed in purified paclitaxel from these two steps, all impurities at or above 0.1% were identified. Results provide information for improving the quality control of paclitaxel production.

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Regeneration of adventitious root from Calystegia soldanella L. in Jeju island and mass proliferation method using bioreactor system (제주지역 갯메꽃(Calystegia soldanella L.) 유래 부정근 재분화 및 생물반응기 시스템 이용 대량증식법)

  • Jong-Du Lee;Eunbi Jang;Weon-Jong Yoon;Yong-Hwan Jung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.37-37
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    • 2021
  • Calystegia soldanella L. is a perennial herbaceous halophyte belonging to the convolvulaceae family, which mainly grows in coastal sand dunes in Korea. Shoots and rhizomes are edible, and roots called 'Hyoseon Chogeun' are known to have medicinal effects such as antipyretic, sterilization, and diuretic. In addition, physiological activities of antioxidant, anti-inflammatory, antiviral, antifungal and PTP-1B (protein tyrosine phosphate-1B) inhibition have been reported. In this study, in vitro induction cell lines of C. soldanella L. collected from the coastal sand dunes in Jeju island was redifferentiated into adventitious roots that can be used as medicinal resources. Also the biomass of mass-proliferated adventitious roots using a bioreactor were evaluated. Plants of C. soldanella L. were collected from the crevice of the seashore in the coastal area of Taeheung 2-ri, Namwon-eup, Seogwipo-si. Then, it was separated into leaves, stems, rhizomes, and roots, and surface sterilized with 70% ethyl alcohol and 2% NaOCl (sodium hypochlorite). After washing with sterilized water, each organ section was cultured in Hormone-free MS medium (Murashige & Skoog Medium). As a result, the induction response rates were evaluated at 85% and 55%, respectively, in terms of callus formation and shoot generation in the rhizome segment. In the case of the adventitious roots morphological characteristics induced by single-use treatment of auxin-based plant growth regulators IBA and NAA from redifferentiated shoots were compared. Most efficient adventitious root culture method as a rooting rate, number, length, and biomass proliferation in the bioreactor system was confirmed when treated by culturing in MS salts, Sucrose 30 g·L-1, and IBA 1mg·L-1 for 4 weeks. In this study, the medium composition and culture period were confirmed using a bioreactor system to mass-proliferate adventitious roots derived from C. soldanella L. in Jeju island. Also this adventitious root line developed a new medicinal material could increase value of the bio-industry ingredient through quantitative and qualitative screening of phyto-bioactive compounds.

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ELECTROPHYSIOLOGICAL AND MORPHOLOGICAL CHARACTERISTICS OF FACIAL NUCLEUS IN RAT (흰쥐 안면신경핵 세포의 전기생리학적 및 형태학적 특성)

  • Choi, Byung-Ju;Cho, Jin-Hwa;Bae, Yong-Chul;Kim, Young-Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.3
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    • pp.400-409
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    • 2000
  • This study used in vivo intracellular and extracellular field potential recording to evaluate the intrinsic membrane properties and connection pattern within facial nucleus. 1. There were four subdivisions of medial, intermediate, lateral, and dorsolateral in facial nucleus. 2. Principal cells in the facial nucleus was recorded from and filled with neurobiotin in anesthetized rats. The extent of their dendrites and the characteristics of cell body were examined. 3. Principal cells had a large amplitude action potential and afterhyperpolarization was followed a single action potential. 4. The response from facial motonucleus to electrical stimulation of the facial nerve was mainly a monophasic wave, with a latency of 1 msec, which was assumed to reflect antidromic activation of facial motoneurons. In some of rats the response in addition showed late components at a latency of about 7-8 msec, but its amplitude was small. 5 Most of cells exhibited accommodation of spike discharge upon depolarization of membrane by 0.8 nA for 400 ms. Our results support the hypothesis that there normally are weak connections between different parts of the facial motonucleus to explain pathophysiology of hemifacial spasm and facial naive paralysis.

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Identification of Chloride Channels in Hamster Eggs (햄스터 난자에서 존재하는 Chloride 통로)

  • Kim, Y.-M.;Kim, J.-S.;Hong, S.-G.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.101-112
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    • 2004
  • Chloride($Cl^-$) channels play critical roles in cell homeostasis and its specific functions such as volume regulation, differentiation, secretion, and membrane stabilization. The presence of these channels have been reported in all kinds of cells and even in frog oocytes. These essential role of $Cl^-$­ channels in cell homeostasis possibly play any role in egg homeostasis and in the early stage of development, however, there has been no report about the presence of $Cl^-$­ channel in the mammalian oocyte. This study was performed to elucidate the presence of $Cl^-$­ channels in hamster eggs. When allowing only $Cl^-$­ to pass through the channel of the egg membrane by using impermeant cation such as N-methyl-D-glucamine(NMDG), single channel currents were recorded. These channel currents showed typical long-lasted openings interrupted by rapid flickering. Mean open $time({\tau}o)$ was 43${\pm}$10.14 ms(n=9, at 50 mV). The open probability(Po) was decrease with depolarization. The current-voltage relation showed outward rectification. Outward slop conductance(32${\pm}$5.4 pS, n=22) was steeper than the inward slop conductance(10${\pm}$1.3 pS). Under the condition of symmetrical 140 mM NaCl, single channel currents were reversed at 0 mV(n=4). This reversal potential(Erev) was shifted from 0 mV at 140 mM concentration of internal NaCl(140 mM [Na+]i) to ­9.8${\pm}$0.5 mV(n=4) at 70 mM [Na+]i and 11.5${\pm}$1.9 mV at 280 mM [Na+]i(n=4) respectively, strongly suggesting that these are single $Cl^-$­ channel currents. To examine further whether this channel has pharmacological property of the $Cl^-$­ channel, specific Cl­ channel blockers, IAA-94(Indanyloxyacetic acid-94) and DIDS(4, 4'-diisothiocyan ostillben- 2-2'disulfonic acid) were applied. IAA-94 inhibited the channel current in a dose-dependent manner and revealed a rapid and flickering block. From these electrophysiological and pharmacological resluts, we found the novel $Cl^-$­ channel present in the hamster oocyte membrane. The first identification of $Cl^-$­ channel in the hamster oocyte may give a clue for the further study on the function of $Cl^-$­ channel in the fertilization and cell differentiation.

[$Na^+-Ca^{2+}$ Exchange Curtails $Ca^{2+}$before Its Diffusion to Global $Ca^{2+}{_i}$ in the Rat Ventricular Myocyte

  • Ahn, Sung-Wan;Ko, Chang-Mann
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.2
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    • pp.95-101
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    • 2005
  • In the heart, $Na^{+}-Ca^{2+}$ exchange (NCX) is the major $Ca^{2+}$ extrusion mechanism. NCX has been considered as a relaxation mechanism, as it reduces global $[Ca^{2+}]_i$ raised during activation. However, if NCX locates in the close proximity to the ryanodine receptor, then NCX would curtail $Ca^{2+}$ before its diffusion to global $Ca^{2+}_i$ This will result in a global $[Ca^{2+}]_i$ decrease especially during its ascending phase rather than descending phase. Therefore, NCX would decrease the myocardial contractility rather than inducing relaxation in the heart. This possibility was examined in this study by comparing NCX-induced extrusion of $Ca^{2+}$ after its release from SR in the presence and absence of global $Ca^{2+}_i$ transient in the isolated single rat ventricular myocytes by using patch-clamp technique in a whole-cell configuration. Global $Ca^{2+}_i$ transient was controlled by an internal dialysis with different concentrations of BAPTA added in the pipette. During stimulation with a ramp pulse from +100 mV to -100 mV for 200 ms, global $Ca^{2+}_i$ transient was suppressed only mildly, and completely at 1 mmol/L, and 10 mmol/L BAPTA, respectively. In these situations, ryanodine-sensitive inward NCX current was compared using $100{\mu}mol/L$ ryanodine, $Na^+$ depletion, 5 mmol/L $NaCl_2$ and $1{\mu}mol/L$ nifedipine. Surprisingly, the result showed that the ryanodine-sensitive inward NCX current was well preserved after 10 mmol/L BAPTA to 91 % of that obtained after 1 mmol/L BAPTA. From this result, it is concluded that most of the NCX-induced $Ca^{2+}$ extrusion occurs before the $Ca^{2+}$ diffuses to global $Ca^{2+})i$ in the rat ventricular myocyte.

Mapping, Tissue Distribution and Polymorphism of Porcine Retinol Binding Protein Genes (RBP5 and RBP7)

  • Gong, W.H.;Tang, Z.L.;Han, J.L.;Yang, S.L.;Wang, H.;Li, Y.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.11
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    • pp.1544-1550
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    • 2008
  • The retinoids (vitamin A and its derivatives) play a critical role in vision, growth, reproduction, cell differentiation and embryonic development. Using the IMpRH panel, porcine cellular retinol binding protein genes 5 and 7 (RBP5 and RBP7) were assigned to porcine chromosomes 5 and 6, respectively. The complete coding sequences (CDS) of the RBP5 and RBP7 genes were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) method, and the deduced amino acid sequences of both genes were compared to human corresponding proteins. The mRNA distributions of the two genes in adult Wuzhishan pig tissues (lung, skeletal muscle, spleen, heart, stomach, large intestine, lymph node, small intestine, liver, brain, kidney and fat) were examined. A total of nine single nucleotide polymorphisms (SNPs) were identified in two genes. Three of these SNPs were analyzed using the polymerase chain reaction-restriction-fragment length polymorphism (PCR-RFLP) method in Laiwu, Wuzhishan, Guizhou, Bama, Tongcheng, Yorkshire and Landrace pig breeds. Association analysis of genotypes of these SNP loci with economic traits was done in our experimental populations. Significant associations of different genotypes of $RBP5-A/G^{63}$, $RBP5-A/G^{517}$ and $RPB5-T/C^{intron1-90}$ loci with traits including maximum carcass length (LM), minimum carcass length (LN), marbling score (MS), back fat thickness at shoulder (SBF), meat color score (MCS) and hematocrit (HCT) were detected. These SNPs may be useful as genetic markers in genetic improvement for porcine production.

New hydroperoxylated and 20,24-epoxylated dammarane triterpenes from the rot roots of Panax notoginseng

  • Shang, Jia-Huan;Sun, Wen-Jie;Zhu, Hong-Tao;Wang, Dong;Yang, Chong-Ren;Zhang, Ying-Jun
    • Journal of Ginseng Research
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    • v.44 no.3
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    • pp.405-412
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    • 2020
  • Background: Root rot is a serious destructive disease of Panax notoginseng, a famous cultivated araliaceous herb called Sanqi or Tianqi in Southwest China. Methods: The chemical substances of Sanqi rot roots were explored by chromatographic techniques. MS, 1D/2D-NMR, and single crystal X-ray diffraction were applied to determine the structures. Murine macrophage RAW264.7 and five human cancer cell lines were used separately for evaluating the antiinflammatory and cytotoxic activities. Results and Conclusion: Thirty dammarane-type triterpenes and saponins were isolated from the rot roots of P. notoginseng. Among them, seven triterpenes, namely, 20(S)-dammar-25-ene-24(S)-hydroperoxyl-3β,6α,12β,20-tetrol (1), 20(S)-dammar-3-oxo-23-ene-25-hydroperoxyl-6α,12β,20-triol (2), 20(S)-dammar-12-oxo-23-ene-25-hydroperoxyl-3β,6α,20-triol (3), 20(S)-dammar-3-oxo-23-ene-25-hydroperoxyl-12β,20-diol (4), 20(S),24(R)-epoxy-3,4-seco-dammar-25-hydroxy-12-one-3-oic acid (5), 20(S),24(R)-epoxy-3,4-seco-dammar-25-hydroxy-12-one-3-oic acid methyl ester (6), and 6α-hydroxy-22,23,24,25,26,27-hexanordammar-3,12,20-trione (7), are new compounds. In addition, 12 known ones (12-16 and 19-25) were reported in Sanqi for the first time. The new Compound 1 showed comparable antiinflammatory activity on inhibition of NO production to the positive control, whereas the known compounds 9, 12, 13, and 16 displayed moderate cytotoxicities against five human cancer cell lines. The results will provide scientific basis for understanding the chemical constituents of Sanqi rot roots and new candidates for searching antiinflammatory and antitumor agents.

A Parentage Test using Indel, Microsatellite Markers and Genotypes of MC1R in the Jeju Black Cattle Population (제주 흑우 집단에서 Indel, Microsatellite 마커와 MC1R 유전자형을 이용한 친자 확인)

  • Han, Sang Hyun;Cho, Sang-Rae;Cho, In-Cheol;Cho, Won-Mo;Kim, Sang-Geum;Yang, Sung-Nyun;Kang, Yong-Jun;Park, Yong-Sang;Kim, Young-Hoon;Park, Se-Phil;Kim, Eun-Young;Lee, Sung-Soo;Ko, Moon-Suck
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.207-213
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    • 2013
  • This study was carried out to examine a molecular marker system for parentage test in Jeju Black cattle (JBC). Based on the preliminarily studies, we finally selected for construction of a novel genetic marker system for molecular traceability, identity test, breed certification, and parentage test in JBC and its related industrial populations. The genetic marker system had eight MS markers, five indel markers, and two single nucleotide polymorphisms (SNPs; g.G299T and g.del310G) within MC1R gene which is critical to verify the breed specific genotypes for coat color of JBC differing from those of exotic black cattle breeds such as Holstein and Angus. The results showed lower level of a combined non-exclusion probability for second parent (NE-P2) of $4.1202{\times}10^{-4}$ than those previously recommended by International Society of Animal Genetics (ISAG) of $5.000{\times}10^{-4}$ for parentage, and a combined non-exclusion probability for sib identity (NE-SI) of $2.679{\times}10^{-5}$. Parentage analysis has been successfully identified the JBC offspring in the indigenous population and cattle farms used the certified AI semens for production using the JBC-derived offspring for commercial beef. This combined molecular marker system will be helpful to supply genetic information for parentage test and traceability and to develop the molecular breeding system for improvement of animal productivity in JBC population.