• Title/Summary/Keyword: CaMS

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The Altered Signaling on EFS-Induced Colon Contractility in Diabetic Rats

  • Thein, Wynn;Po, Wah Wah;Kim, Dong Min;Sohn, Uy Dong
    • Biomolecules & Therapeutics
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    • v.28 no.4
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    • pp.328-336
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    • 2020
  • Diabetes mellitus affects the colonic motility developing gastrointestinal symptoms, such as constipation. The aim of the study was to examine the role of intracellular signaling pathways contributing to colonic dysmotility in diabetes mellitus. To generate diabetes mellitus, the rats were injected by a single high dose of streptozotocin (65 mg/kg) intraperitoneally. The proximal colons from both normal and diabetic rats were contracted by applying an electrical field stimulation with pulse voltage of 40 V in amplitude and pulse duration of 1 ms at frequencies of 1, 2, 4, and 6 Hz. The muscle strips from both normal rats and rats with diabetes mellitus were pretreated with different antagonists and inhibitors. Rats with diabetes mellitus had lower motility than the control group. There were significant differences in the percentage of inhibition of contraction between normal rats and rats with diabetes mellitus after the incubation of tetrodotoxin (neuronal blocker), atropine (muscarinic receptor antagonist), prazosin (α1 adrenergic receptor antagonist), DPCPX (adenosine A1 receptor antagonist), verapamil (L-type Ca2+ channel blocker), U73122 (PLC inhibitor), ML-9 (MLCK inhibitor), udenafil (PDE5 inhibitor), and methylene blue (guanylate cyclase inhibitor). The protein expression of p-MLC and PDE5 were decreased in the diabetic group compared to the normal group. These results showed that the reduced colonic contractility resulted from the impaired neuronal conduction and decreased muscarinic receptor sensitivity, which resulted in decreased phosphorylation of MLC via MLCK, and cGMP activity through PDE5.

The Role of Adenosine Receptor on Norepinephrine Release from Ischemic-Induced Rat Hippocampus (허혈이 유발된 흰쥐 해마에서 Norepinephrine 유리에 미치는 Adenosine 수용체의 역할)

  • Chung, Jong-Hoon;Choi, Bong-Kyu
    • The Korean Journal of Pharmacology
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    • v.32 no.2
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    • pp.139-150
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    • 1996
  • The effects of adenosine analogues on the electrically-evoked norepinephrine(NE) release and the influence of ischemia on the effects were studied in the rat hippocampus. Slices from the rat hippocampus were equilibrated with $0.1{\mu}M$ $[^3H]-norepinephrine$ and the release of the labelled product, $[^3H]-NE$, was evoked by electrical stimulation(3 Hz, 2 ms, 5 $VCm^{-1}$ and rectangular pulses for 90 sec), and the influence of various agents on the evoked tritium-outflow was investigated. Ischemia(15min with 95% $N_2$ +5% $CO_2$) increased both the basal and evoked NE release. These increases were abolished by addition of glucose into the superfused medium, and they were significantly inhibited either by $0.3\;{\mu}M$ tetrodotoxin pretreatment or by removing $Ca^{++}$ in the medium. MK-801$(1{sim}10\;{\mu}M)$, a specific NMDA receptor antagonist, and glibenclamide $(1\;{\mu}M)$, a $K^+-channel$ inhibitor, neither alter the evoked NE release nor affected the Ischemia-Induced increases in NE release. However, polymyxin B(0.03 mg), a specific protein kinase C inhibitor, inhibited the effect of ischemia on the evoked NE release. Adenosine and $N^6-cyclopentyladenosine$ decreased the NE release in a dose-dependent manner in ischemic condition, though the magnitude of inhibition was far less than those in normal (normoxic) condition. Also the treatment with $5{\mu}M$ DPCPX, a potent $A_1-adenosine$ receptor antagonist did not affect the ischemia-effect. These results suggest that the evoked-NE release is potentiated by ischemia, and this process being most probably mediated by protein kinase C, and that the decrease of NE release mediated through $A_1-adenosine$ receptor is significantly inhibited in ischemic state.

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InhA-Like Protease Secreted by Bacillus sp. S17110 Inhabited in Turban Shell

  • Jung, Sang-Chul;Paik, Hyoung-Rok;Kim, Mi-Sun;Baik, Keun-Sik;Lee, Woo-Yiel;Seong, Chi-Nam;Choi, Sang-Ki
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.402-408
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    • 2007
  • A strain producing a potent protease was isolated from turban shell. The strain was identified as Bacillus sp. S17110 based on phylogenetic analysis. The enzyme was purified from culture supernatant of Bacillus sp. S17110 to homogeneity by ammonium sulfate precipitation, SP-Sepharose, and DEAE-Sepharose anion exchange chromatography. Protease activity of the purified protein against casein was found to be stable at pH 7 to pH 10 and around $50^{\circ}C$. Approximately 70% of proteolytic activity of the enzyme was detected either in the presence of 100 mM SDS or Tween 20. The enzyme activity was enhanced in the presence of $Ca^{2+},\;Zn^{2+},\;Mg^{2+}$, but was inhibited by EDTA, indicating that it requires metal for its activity. The purified enzyme was found to be a monomeric protein with a molecular mass of 75 kDa, as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography. The purified enzyme was analyzed through peptide fingerprint mass spectra generated from matrix-assisted laser desorption ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) and a BLAST search, and identified as immune inhibitor A (inhA) deduced from nucleotide sequence of B. cereus G9241. Since InhA was identified as protease that cleave antibacterial proteins found in insect, inhA-like protease purified from Bacillus sp. S17110 might be pathogenic to sea invertebrates.

Influence of Essential Oil in 'Shiranuhi' Immature Fruit on Antioxidant and Antimicrobial Activities (부지화 미숙과 에센셜 오일의 항산화 및 항균 활성 효과)

  • Kim, Sang Suk;Hyun, Ju Mi;Kim, Kwang Sik;Park, Kyung Jin;Park, Suk Man;Choi, Young Hun
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.6
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    • pp.493-497
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    • 2013
  • This study was designed to analyze the chemical composition of essential oil in 'Shiranuhi' immature fruit and to test their biological activities. 'Shiranuhi' immature essential oils (SIEO) were obtained by steam distillation from fruits collected from Jeju Island and were analyzed using gas chromatograph (GC)-flame ionization detectors (FID) and GC-MS. Fourteen components were identified in the essential oil. Limonene (75.21%) and terpineol (8.68%) were the major components in SIEO. Since acne vulgaris is the combined result of a bacterial infection and the inflammatory response to that infection, we examined whether SIEO possessed antibacterial against skin pathogens. As a result, SIEO showed excellent antibacterial activities against drug-susceptible and -resistant Propionibacterium acnes and Staphylococcus epidermidis, which are acne-causing bacteria. In this study, SIEO was examined on DPPH radical scavenging activities, which showed moderate antioxidant activity ($SC_{50}$, $15.36{\mu}L/mL$). In order to determined whether SIEO can be safely applied to human skin, the cytotoxicity effects of SIEO were determined by colorimetric MTT assays in normal human fibroblasts and keratinocyte HaCaT cells. They exhibited low cytotoxicity at $0.5{\mu}L/mL$ in both celllines. Based on these results, we suggest the possibility that essential oil of 'Shiranuhi' maybe considered as an antibacterial and antioxidant agent.

Thiol-dependent Redox Mechanisms in the Modification of ATP-Sensitive Potassium Channels in Rabbit Ventricular Myocytes

  • Han, Jin;Kim, Na-Ri;Cuong, Dang-Van;Kim, Chung-Hui;Kim, Eui-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.1
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    • pp.15-23
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    • 2003
  • Cellular redox state is known to be perturbed during ischemia and that $Ca^{2+}$ and $K^2$ channels have been shown to have functional thiol groups. In this study, the properties of thiol redox modulation of the ATP-sensitive $K^2$ ($K_{ATP}$) channel were examined in rabbit ventricular myocytes. Rabbit ventricular myocytes were isolated using a Langendorff column for coronary perfusion and collagenase. Single-channel currents were measured in excised membrane patch configuration of patch-clamp technique. The thiol oxidizing agent 5,5'-dithio-bis-(2-nitro-benzoic acid) (DTNB) inhibited the channel activity, and the inhibitory effect of DTNB was reversed by dithiothreitol (disulfide reducing agent; DTT). DTT itself did not have any effect on the channel activity. However, in the patches excised from the metabolically compromised cells, DTT increased the channel activity. DTT had no effect on the inhibitory action by ATP, showing that thiol oxidation was not involved in the blocking mechanism of ATP. There were no statistical difference in the single channel conductance for the oxidized and reduced states of the channel. Analysis of the open and closed time distributions showed that DTNB had no effect on open and closed time distributions shorter than 4 ms. On the other hand, DTNB decreased the life time of bursts and increased the interburst interval. N-ethylmaleimide (NEM), a substance that reacts with thiol groups of cystein residues in proteins, induced irreversible closure of the channel. The thiol oxidizing agents (DTNB, NEM) inhibited of the $K_{ATP}$ channel only, when added to the cytoplasmic side. The results suggested that metabolism-induced changes in the thiol redox can also modulate $K_{ATP}$ channel activity and that a modulatory site of thiol redox may be located on the cytoplasmic side of the $K_{ATP}$ channel in rabbit ventricular myocytes.

Detrital zircon U-Pb ages of the uppermost Jinju Formation in the Natural Monument No. 534 'Tracksite of Pterosaurs, Birds, and Dinosaurs in Hotandong, Jinju', Korea

  • Chae, Yong-Un;Lim, Jong Deock;Kim, Cheong-Bin;Kim, Kyung Soo;Ha, Sujin;Lim, Hyoun Soo
    • Journal of the Korean earth science society
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    • v.41 no.4
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    • pp.367-380
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    • 2020
  • LA-MC-ICP-MS zircon U-Pb dating was conducted to constrain the timing of fossil formation and the depositional age of the uppermost Jinju Formation located in Natural Monument No. 534 (Tracksite of Pterosaurs, Birds, and Dinosaurs in Hotandong, Jinju), and 87 Cretaceous, 1 Precambrian, and 2 Jurassic zircons were obtained from 90 valid analytical points. Most Cretaceous zircons were found to have a youngest graphical peak age of ca. 106.5 Ma, suggesting the depositional age of the uppermost Jinju Formation. Based on this study and previous works, the average sedimentation rate of the Jinju Formation was calculated to be approximately 0.17-0.31 mm/year in the Milyang Subbasin, and the Cretaceous zircons of the uppermost Jinju Formation seem to have originated mainly from the western or northwestern parts of the Gyeonggi Massif. Unlike the Nakdong and Hasandong formations of the Sindong Group, most zircons analyzed in the uppermost Jinju Formation were Cretaceous. This suggests that volcanic activity occurred in the area closer to the Gyeongsang Basin due to the roll-back of subducting paleo-Pacific Plates during the Jinju period.

Physiological and Proteomics Analysis to Potassium Starvation in Rice

  • Kim, Sang-Gon;Wang, Yiming;Lee, Chang-Hoon;Chi, Yong-Hun;Kim, Keun-Ki;Choi, In-Soo;Kim, Yong-Chul;Kang, Kyu-Young;Kim, Sun-Tae
    • Korean Journal of Environmental Agriculture
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    • v.30 no.4
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    • pp.395-401
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    • 2011
  • BACKGROUND: Potassium (K) is one of the macronutrients which are essential for plant growth and development. Its deficiency in paddy soils is becoming one of the limiting factors for increasing rice yield in Asia. METHODS AND RESULTS: To investigate physiological symptoms under K-starvation (NP) compared with complete media (NPK) condition, we measured shoot/root length, weight, nutrients, and patterns of protein expression. The shoot growth was significantly reduced, but root growth was not affected by K-starvation. However, biomasses were decreased in both shoot and root. Uptake of K was reduced up to 85%, while total concentrations of P, Ca, Mg, Na were increased in root and shoot. To better understand the starved K mechanism of rice, comparative proteome analysis for proteins isolated from rice leaves was conducted using 2-DGE. Five spots of differentially expressed proteins were analyzed by MALDI-TOF MS. Analysis of these K-starvation response proteins suggested that they were involved in metabolism and defense. CONCLUSION(s): Physiological and 2-DGE based proteomics approach used in our study results in observation of morphology or nutrients change and identification of K-starvation responsive proteins in rice root. These results have important roles in maintaining nutrient homeostasis and would also be useful for further characterization of protein function in plant K nutrition.

Effect of Protein Kinase C on Norepinephrine Release in the Rat Hippocampus (흰쥐 해마에서 Norepinephrine 유리에 미치는 Protein Kinase C 의 영향)

  • Kim, Do-Kyung;Lee, Young-Soo;Choi, Bong-Kyu
    • The Korean Journal of Pharmacology
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    • v.31 no.2
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    • pp.145-152
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    • 1995
  • The effects and interactions of $4{\beta}-phorbol$ 12,13-dibutyrate(PDB) and polymyxin B(PMB) with adenosine on the electrically-evoked norepinephrine (NE) release were studied in the rat hippocampus. Slices from the rat hippocampus were equilibrated with $^3H-noradrenaline$ and the release of the labelled product, $^3H-NE$, which evoked by electrical stimulation$(3\;Hz,\;2\;ms,\;5\;VCm^{-1},\;rectangular\;pulses)$ was measured. PDB$(0.3{\sim}10\;{\mu}M)$, a selective protein kinase C(PKC) activator, increased the evoked NE release in a dose related fashion while increasing the basal rate of release. And the effects of $1\;{\mu}M$ PDB were significantly inhibited by $0.3\;{\mu}M$ tetrodotoxin(TTX) pretreatment or $Ca^{++}-free$ medium. $PMB(0.03{\sim}1\;mg)$, a specific PKC inhibitor, decreased the NE release in a dose dependent manner while increasing the basal rate of release. Adenosine $(1{\sim}10\;{\mu}M)$ decreased the NE release without changing the basal rate of release, and this effect was significantly inhibited by 8-cyclopentyl-1,3-dipropylxanthine$(2\;{\mu}M)$, a selective $A_1-receptor$ antagonist, treatment. Also, adenosine effects were significantly inhibited by PDB-and PMB-pretreatment. These results suggest that the PKC plays a role in the NE release in the rat hippocampus and might be participated in a post-receptor mechanism of the $A_1-adenosine$ receptor.

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Study on Thermal Properties of CdS - Embedded Poly(2-Acetamidoacrylic acid) Hydrogel Composite (CdS 나노입자틀 삽입한 Poly(2-Acetamidoacrylic acid) 수화젤 복합체의 열적 특성에 관한 연구)

  • Park, Chun-Ho;Ha, Eun-Ju;Jung, Jong-Mo;Lee, Jang-Oo;Paik, Hyun-Jong
    • Polymer(Korea)
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    • v.33 no.1
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    • pp.1-6
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    • 2009
  • We report the template-based synthesis of well-dispersed CdS nanoparticles (NPs) in the interior of poly (2-acetamidoacrylic acid) (PAAA) hydrogel as a novel type of nanocomposite without particle aggregation via ion exchange in a aqueous system. As revealed by the TEM image analysis, the mean crystallite diameter of CdS NPs embedded in hydrogel composite was 4.5 nm, and the composite did not suffer any observable change after 6 months. Desorption/decomposition of CdS/PAAA hydrogel composite was studied by evolved gas analysis-gas chromatography-mass spectrometry (EGA-GC-MS), and thermogravimetric analysis (TGA) methods. From the TGA data, the thermal stability of the composite system increased by ca. 100 $^\circ$C and the content of CdS NPs in a dry composite gel was over 70 wt%. In addition, the chemical pathway was proposed for the entire decomposition process.

A Study on the Characteristics and Burial Age of Sediment Layers at Bukpyeong myeon, Haenam gun (해남 북평면 퇴적층의 특성과 매몰 연대에 대한 연구)

  • Shin, Won Jeong;Yang, Dong Yoon;Kim, Jong Yeon
    • Journal of The Geomorphological Association of Korea
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    • v.23 no.3
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    • pp.41-55
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    • 2016
  • The granulometric characteristics and chemical composition of the samples from the Bukpyeong myeon, Hainam gun, Jeollanam do were analyzed in this study. The samples were collected from the outcrop of coastal area where the height is about 30m. The burial age of the sediments also estimated by OSL(Optically Stimulated Luminescence) method. The origin and forming processes of the samples are not clear in this stage as limited number of evidences can be found. However it is suggested that the samples are not aeolian deposits originated from chinese loess by the chemical analysis. The mean diameter of samples were $5{\sim}6{\varphi}(silt)$ and are increased slightly downwards. The samples also well sorted. The samples have been heavily weathered by the chemical alteration index. The value of CIA increase downward rapidly at the upper part of outcrop, then stabilized afterwards. The vertical concentration changes ratio of cations varies; Si increases downwards while Fe, Ti, K and Mg decrease. There are fluctuation in concentration ratio of Na and Ca with increasing depth. The burial age of sample from upper part are estimated as $104.52{\pm}4.45ka\;BP$, while that of lower part are $136.10{\pm}6.52ka\;BP$, and they fall in to last part of MS6. The deposition rate for this site is found about 0.017mm/year. The uplift rate of the site is supposed to be 0.24~0.26m/ka with assumption of palaeo-sea level of +6m. However the origin of the sediment deposits should be explored and cleared.