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Dual Regulation of R-Type CaV2.3 Channels by M1 Muscarinic Receptors

  • Jeong, Jin-Young;Kweon, Hae-Jin;Suh, Byung-Chang
    • Molecules and Cells
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    • v.39 no.4
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    • pp.322-329
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    • 2016
  • Voltage-gated $Ca^{2+}$ ($Ca_V$) channels are dynamically modulated by Gprotein-coupled receptors (GPCR). The $M_1$ muscarinic receptor stimulation is known to enhance $Ca_V2.3$ channel gating through the activation of protein kinase C (PKC). Here, we found that $M_1$ receptors also inhibit $Ca_V2.3$ currents when the channels are fully activated by PKC. In whole-cell configuration, the application of phorbol 12-myristate 13-acetate (PMA), a PKC activator, potentiated $Ca_V2.3$ currents by ~two-fold. After the PMA-induced potentiation, stimulation of $M_1$ receptors decreased the $Ca_V2.3$ currents by $52{\pm}8%$. We examined whether the depletion of phosphatidylinositol 4,5-bisphosphate ($PI(4,5)P_2$) is responsible for the muscarinic suppression of $Ca_V2.3$ currents by using two methods: the Danio rerio voltage-sensing phosphatase (Dr-VSP) system and the rapamycin-induced translocatable pseudojanin (PJ) system. First, dephosphorylation of $PI(4,5)P_2$ to phosphatidylinositol 4-phosphate (PI(4)P) by Dr-VSP significantly suppressed $Ca_V2.3$ currents, by $53{\pm}3%$. Next, dephosphorylation of both PI(4)P and $PI(4,5)P_2$ to PI by PJ translocation further decreased the current by up to $66{\pm}3%$. The results suggest that $Ca_V2.3$ currents are modulated by the $M_1$ receptor in a dual mode-that is, potentiation through the activation of PKC and suppression by the depletion of membrane $PI(4,5)P_2$. Our results also suggest that there is rapid turnover between PI(4)P and $PI(4,5)P_2$ in the plasma membrane.

Optimization of TiCl4 Concentration and Initial pH for Phosphorus Removal in Synthetic Wastewater (합성폐수 내 인을 제거하기 위한 TiCl4 농도 및 초기 pH 최적조건 도출)

  • Shin, So-Yeun;Kim, Jong-Ho;Ahn, Johng-Hwa
    • Journal of Korean Society on Water Environment
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    • v.31 no.6
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    • pp.619-624
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    • 2015
  • This study experimentally determined the effect of titanium tetrachloride (TiCl4) concentration ([TiCl4]) (0.25-0.55 mM) and initial pH (3-11) on phosphorus (P) removal in synthetic wastewater (2 mg P/L). The P removal efficiency increased when [TiCl4] increased. The P removal efficiency showed a parabolic trend with an inflection point at pH 7. At the molar ratio of TiCl4 and P>6.2, the P removal efficiency was over 90% and the residual P concentration was less than 0.2 mg/L. Within the design boundaries, the complete P removal could be achieved at 7.0≤initial pH≤8.5 and 0.43≤[TiCl4]≤0.55 mM. The final pH was over 5.8 at initial pH≥7.7 and [TiCl4]≥0.35 mM. The results showed that TiCl4 was effective in P removal in water so that it could be an alternative chemical for P removal.

Effects of Propofol on Electroencephalogram in Dogs (Propofol이 개의 뇌파에 미치는 영향)

  • 장환수;장광호;채형규;권은주;김정은
    • Journal of Veterinary Clinics
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    • v.17 no.2
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    • pp.359-367
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    • 2000
  • The aim of this study was to evaluate the effects of propofol on cortical electroencephalogram (EEG) in seven dogs. Propofol infusion was accomplished from low concentration to high concentration in series, and each concentration was infused for 20 minutes (M0: 0, M0.5: 0.5, M1.0:1.0, and M1.5: 1.5 mg/kg/min of infusion rate). EEG was recorded via needle electrode placed at Cz, which was applied to International 10-20 system. Arterial blood pressure. blood gas analysis and ECG were also measured. Hoemodynamics, Pa$CO_2$, PaO$_2$, heart rate and respiratory rate were variable, but were net significant(p>0.05). The power spectra of EEG in every concentration was compared wish those of control (MO). The powers at a1l frequencies at M1.0 and Ml.5 were decreased. Especially, the powers of the frequencies over 20 Hz were significantly decreased (p<0.O5). Powers at frequencies between 8 and 15Hz at MO.S were significantly increased (p<0.05) in response to the painful stimuli. It was inferred that they may reflect activity of the brain which is consciously processing the external Stimuli. Like the Power spectra, al1 the band powers of He EEG ($\delta$ 1-4, $\theta$4-8, $\alpha$ 8-13, $\beta$L13-21. $\beta$H 21-30, \ulcorner 30-50, and total 1-5OHz) were decreased in proportion to the increase of infusion rate at M1 .0 and M1.5. Especially, decrease of $\beta$H and ${\gamma}$ were significant(p<0.01). At M0.5, $\alpha$ band was significantly increased(p<0.05) among all the bands. Seizure activities which were concide with occurrence of spike wave were shown in all dogs at Ml .0 and M1.5.

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Fnr, NarL and NarP Regulation and Time Course Expression of Escherichia coli aeg-46.5 Gene

  • Ahn, Ju-Hyuk;Choe, Mu-Hyeon
    • BMB Reports
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    • v.29 no.1
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    • pp.88-91
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    • 1996
  • The anaerobically expressed gene aeg-46.5, which had been identified by the operon fusion technique with a hybrid bacteriophage of ${\lambda}$ and Mu, ${\lambda}$placMu53, was studied for its expression pattern and growth. The expression of aeg-46.5 was studied in the wild-type cell and mutant cells that have mutation (s) in the control gene of anaerobic respiration (fnr) and nitrate response (narL and narP). The ${\beta}$-galactosidase reporter gene showed maximum expression in narL host after two hours of aerobic to anaerobic switch in M9-Glc-nitrate medium. Both 40 mM and 100 mM concentrations of nitrate ion in the medium had little effect on expression level. We propose that aeg-46.5 is subject to multiple regulations of anaerobic activation by Fnr, nitrate activation by NarP and repression mediated by NarL.

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Effect of P(VDF/TrFE) Film Thickness on the Characteristics of Pyroelectric Passive Infrared Ray Sensor for Human Body Detection (P(VDF/TrFE) 필름의 두께에 따른 인체 감지형 초전형 PIR 적외선 센서의 특성)

  • Kwon, Sung-Yeol
    • Journal of Sensor Science and Technology
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    • v.20 no.2
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    • pp.114-117
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    • 2011
  • A thick 25 ${\mu}m$ thickness poled P(VDF/TrFE) film pyroelectric infrared ray sensor has been fabricated and then thin 1.6 ${\mu}m$ thickness P(VDF/TrFE) film pyroelectric infrared ray sensor has been fabricated also. These thick and thin P(VDF/TrFE) film pyroelectric infrared ray sensor was mounted in TO-5 housing to detect infrared light of 5.5 ~ 14 ${\mu}m$ wavelength for human body detecting with each other. The noise output voltage of the thick P(VDF/TrFE) film pyroelectric infrared ray sensor were 380 mV and NEP(noise equivalent power) is $3.95{\times}10^{-7}$ W which is the similar value with the commercial pyroelectric infrared ray sensor using ceramic materials as a sensing material. The NEP and specific detectivity $D^*$ of the thin P(VDF/TrFE) film pyroelectric infrared ray sensor were $2.13{\times}10^{-8}$ W and $9.37{\times}106$ cm/W under emission energy of 13 ${\mu}W/cm^2$ respectively. These result caused by lower thermal diffusion coefficient of a thin 1.6 ${\mu}m$ thickness PVDF/TrFE film than the thick 25 ${\mu}m$ thickness poled P(VDF/TrFE) film pyroelectric infrared ray sensor.

Characterization of the Restriction Endonuclease Bki I from Brevibacterium divaricatum (Brevibacterium divaricatum의 제한효소 Bdi I의 특성)

  • 김용석;노현모
    • Korean Journal of Microbiology
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    • v.24 no.1
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    • pp.18-23
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    • 1986
  • A new type II restriction endonuclease, Bdi I, has been isolated from Brenibacterium divaricatum FERM 5948 by procedures of ammonium sulfate fractionation, DEAE-cellulose chromatography and heparin agarose chromatography. The purified Bdi I restriction endonudlease had the same cleavage patterns of Cla I whose recognition sequence is 5' ATCGAT 3'. From the result that ${\lambda}-Cla$ I DNA frahment could be cloned in pBR 322 digested with Bdi I, it has been proven that Bdi I cuts between T and C(5' AT/CGAT3') within the recognition sequence and produces 5'pCG cohesive end. The optimal temperature for the Bdi I restriction endonuclease activity was $37^{\circ}C$, and optimal salt (NaCl) concentration was 50-100 mM.

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Inhibitory effects of heavy metals on CYP1A expression in eel hepatocyte cultures (뱀장어 배양 간세포에서의 Cytochrome P4501A (CYP1A) 유전자 발현에 대한 중금속들의 억제효과)

  • Kwon, Hyuk-Chu;Maeng, Joon-Ho;Choi, Seong-Hee
    • Journal of fish pathology
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    • v.23 no.2
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    • pp.245-254
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    • 2010
  • Effects of heavy metal ions on the gene expression of cytochrome P4501A (CYP1A) were examined in cultured eel hepatocytes. When the expression of CYP1A mRNA was measured by RT-PCR after incubation of eel hepatocytes with benzo[$\alpha$]pyrene (B[$\alpha$]P) at concentrations of 10-8~10-5 M, the CYP1A expression increased with B[$\alpha$]P treatment in a dose dependent manner, showing significant increase at concentrations more than 10-7 M. When the eel hepatocyte was treated with cadmium (10-6 and 10-5 M), the expression of CYP1A was inhibited and especially at higher concentration (10-5 M). The inhibition of CYP1A expression by cadmium was also observed in cells treated with B[$\alpha$]P. In another study, effects of heavy metal ions on the expression of CYP1A were examined in cultured hepatocytes isolated from eel which was treated previously with B[$\alpha$]P in vivo. Hepatocytes isolated from the liver of eel taken at 48 hours after injection of B[$\alpha$]P (10 mg/kg) were cultured for 2 days with cadmium, copper, lead or zinc (10-6 and 10-5 M). The expression of CYP1A was found to be suppressed by the metal ions compared with the control in which CYP1A was induced with previous treatment of B[$\alpha$]P in vivo. The present results may provide an important basic information for studying the effects of heavy metal ions on CYP1A expression in other species of fish and studying toxicological mechanisms of heavy metal ions in aquatic livings.

Applicability Assessment of Carbon Nanotube to Slow Sand Filtration for Bacteria Removal (박테리아 제거를 위한 완속 모래여과에서 탄소나노튜브의 적용성 검토)

  • An, Hee-Kyung;Park, Seong-Jik
    • Journal of Korean Society of Environmental Engineers
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    • v.36 no.12
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    • pp.873-878
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    • 2014
  • The applicability of carbon nanotube (CNT) to slow sand filtration for the removal of bacteria was studied using scanning electron microscope and column experiments. The morphology of CNT were investigated using scanning electron microscope and the CNT looked like a skein serving bacteria favorable site for adhesion. Column experiments were performed over a range of CNT filter depth, pH, and ionic strength. Bacteria removal efficiency was found to increase from 44.15% to 99.95% as the CNT filter depth increased from 1 cm to 5 cm, and 3 cm of CNT filter depth was required for significant removal of bacteria. pH increase from 5.5 to 8.5 decreased the bacteria removal efficiency, due to the electrostatic repulsion between bacteria and CNT at higher pH. Bacteria removal efficiency decreased from 97.25% to 70.90% as the ionic strength increased from 0 mM to 50 mM. This study demonstrated that the CNT can be applied to slow sand filtration for treating microbially contaminated water.

Interactions between Hydrodenitrogenation of Pyridine and Hydrodeoxygenation of m-Cresol over sulfided CoMo/γ-Al2O3 Catalyst (황화 CoMo/γ-Al2O3 촉매상에서 수첨탈질반응과 수첨탈산소 반응의 상호작용)

  • Kim, Hak-Soo;Park, Hea-Kyung;Kim, Kyung-Lim
    • Applied Chemistry for Engineering
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    • v.2 no.2
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    • pp.108-118
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    • 1991
  • Interactions between pyridine hydrodenitrogenation (HDN) and m-cresol hydrodeoxygenation(HDO), and the kinetic analysis were studied over sulfided $CoMo/{\gamma}-Al_2O_3$ catalyst at the range of temperatures between 473 K and 723 K, the total pressures between $10{\times}10^5Pa$ and $50{\times}10^5Pa$, and the contact times between 0.0125 g-cat. hr/ml-feed and 0.03g-cat. hr/ml-feed. HDN of pyridine and HDO of m-cresol were inhibited by each other and the inhibition effect of HDO by pyridine is higher than that of HDN by m-cresol. But reactivity of m-cresol is higher than that of pyridine. The rate equations of pyridine and m-cresol were given to be ${\gamma}_{HDN}=k_{HDN}{\cdot}K_pC_p/(1+K_cC_c+K_pC_p)$ and ${\gamma}_{HDO}=k_{HDO}{\cdot}K_cC_c/(1+K_cC_c+K_pC_p)$ in terms of Langmuir-Hinshellwood-Hougen-Watson model. At each temperature, reaction rate constants and adsorption equilibrium constants were determined and activation energies of pyridine HDN and m-cresol HDO are 13.83kcal/mol, respectively and the heat of adsorption are -6.458 and -5.045kcal/mol, respectively.

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Purification and some properties of polyphenol oxidase from Spuriopimpinella bracycarpa (참나물로부터 추출한 polyphenol oxidase의 부분정제 및 성질)

  • Ham, Seung-Shi;Hong, Eun-Hee;Lee, Sang-Young;Park, Gwi-Gun;Omura, Hirohisa
    • Applied Biological Chemistry
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    • v.34 no.1
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    • pp.49-53
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    • 1991
  • Three polyphenol oxidase(polyphenol oxidase I, II and III ) were isolated from the crude extract of a Spuriopimpinella bracycarpa by $(NH_4)_2SO_4$ precipitation and subsequent Sephadex G-150 chromatography. The final preparation thus obtained showed three peaks of enzyme activity. Optimum pH and temperature for the activity of polyphenol oxidase were 7.5 and $30^{\circ}C$, respectively. The enzyme was completely inactivated when i4 was treated at$70^{\circ}C$ for 30min and at $80^{\circ}C$ for 5min at pH 6.5. The enzyme was partially inactivated by ascorbic acid, glutathione and potassium cyanide (0.1mM), and was completely inhibited by L-cysteine, ascorbic acid, glutathione and potassium cyanide(0.5 and 1.0mM). The enzyme has good activity on catechol and 3,4-dihydroxytoluene but was strongly inactivated on pyrogallol, dopamine and DL-dopa. The Michaelis cons4ant of the enzyme was 86.5mM with catechol as a substrate.

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