The Journal of Korean Institute of Communications and Information Sciences
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v.18
no.4
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pp.468-480
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1993
In this paper, at first, the characteristics of double mode SAW (DMS) resonator are analysed by applying the wave guide model and coupled mode theory to 1-port resonator. The DMS resonators (2-pole), which are arranged in a close, parallel configuration of two identical 1-port resonators on a single plate, and 4-pole DMS filters are designed and fabricated at the center frequency of 150.15 MHz with 3-dB bandwidth of 80 KHz. The empirical design characteristics are obtained from the comparison of experimental and theoretical values resulted from several fabrications, and the narrow bandpass filters are implemented on the basis of the above empirical results, which can be used to mobile communication systems. A ST-cut quartz substrate is selected for the stable temperature-frequence characteristics, and high resolution photolithography is applied to the fabrication of filter to get the fine electrode patterns.
Objectives: Metabolic syndrome is correlated with increased cardiovascular risk and characterized by several factors, including visceral obesity, hypertension, insulin resistance, and dyslipidemia. Several members of a large family of nonselective cation entry channels, e.g., transient receptor potential (TRP) melastatin 7 (TRPM7) channels have been associated with the development of cardiovascular diseases. The purpose of this study was to investigate the effects of Dangkwisoo-san, ginger and curcumin on TRPM7 channel. Methods: Human embryonic kidney (HEK) 293 cells stably transfected with the TRPM7 expression vectors were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, $5{\mu}g/mL$ blasticidin, and 0.4 mg/mL zeocin in a humidified 20% $O_2$/10% $CO_2$ atmosphere at $37^{\circ}C$. Whole-cell patch clamp recordings were obtained using an Axopatch 700B amplifier and pClamp v.10.4 software, and signals were digitalized at 5 kHz using Digidata 1422A. Results: Dangkwisoo-san extract (100, 200, 300, 400, and $500{\mu}g/mL$) inhibited the outward and inward TRPM7 whole-cell currents at dose dependent manner and the half maximal inhibitory concentration $(IC)_{50}$ of Dangkwisoo-san was $218.3{\mu}g/mL$. Also, ginger extract (100, 200, 300, 400, and $500{\mu}g/mL$) inhibited the outward and inward of TRPM7 whole-cell currents in a dose dependent manner and the $IC_{50}$ of ginger was $877.2{\mu}g/mL$. However, curcumin had no effects on TRPM7 whole-cell currents. Conclusions: These results suggest that both Dangkwisoo-san and ginger have good roles to inhibit the TRPM7 channel, suggesting that Dangkwisoo-san and ginger are considered one of the candidate agents for the treatment of metabolic syndrome such as cardiovascular disease.
Objectives: Metabolic syndrome is defined by a cluster of major cardiovascular risk factors: obesity, insulin resistance, dyslipidemia, and arterial hypertension. Several members of a large family of nonselective cation entry channels, e.g., transient receptor potential vanilloid 4 (TRPV4) channels have been associated with the development of dyslipidemia and hypertension. The purpose of this study was to investigate the effects of Leejung-tang (Lizhong-tang), Rikkunshito, and Bojungikgi-tang (Buzhongyiqi-tang) on TRPV4 channel. Methods: Human embryonic kidney 293 cells stably transfected with the TRPV4 expression vectors were maintained in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin, $5{\mu}g/mL$ blasticidin, and 0.4 mg/mL zeocin in a humidified 20% $O_2/10%$$CO_2$ atmosphere at $37^{\circ}C$. Whole-cell patch clamp recordings were obtained using an Axopatch 700B amplifier and pClamp v.10.4 software (Molecular Devices, San Jose, CA, USA), and signals were digitalized at 5 kHz using Digidata 1422A. Results: Leejung-tang and Rikkunshito (10, 30 and 50 mg/mL) had no effects on the TRPV4 whole-cell currents at dose dependent manner. However, Bojungikgi-tang (10, 30, and 50 mg/mL) inhibited the TRPV4 whole-cell currents in a dose dependent manner and the half maximal inhibitory concentration (IC50) of Bojungikgi-tang was 18.2 mg/mL. Conclusions: These results suggest that Bojungikgi-tang plays an important roles to inhibit the TRPV4 channel, suggesting that Bojungikgi-tang is considered one of the candidate agents for the treatment of metabolic syndrome such as dyslipidemia and hypertension.
The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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v.5
no.3
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pp.208-215
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2000
The estimated total material transports through the Cheju Strait using all data which investigated in 1997 and 1999 are as follows; A large amount of suspended sediments and dissovted inorganic nutrients are carried tothe South Sea through the Cheju Strait by a persistent eastward flow (Cheju Current) from the Y311ow Sea andthe East China Sea. The annual material Oanspous by the Cheju Current are as follows; 22.9${\times}$10$^6$ ton yr$^{-1}$(SS), 0.52${\times}$10$^{10}$ mol yr$^{-1}$ (NH$_4\;^+$), 6.05${\times}$10$^{10}$ mol yr$^{-1}$ (NO$_3\;^-$), 0.36${\times}$10$^{10}$ mol yr$^{-1}$ (PO$_4\;^{3-}$), 10.27${\times}$10$^{10}$ mol yr$^{-1}$ (Si(OH)$_4$). The annual suspended sediment flux per water transport in the Cheju Strait (44.48${\times}$10$^6$ ton yr$^{-1}$ Sv$^{-1}$) is about 1.7 larger than that in the Korean Strait (26.08${\times}$10$^6$ ton yr$^{-1}$ Sv$^{-1}$). The annual nitrate flux per water transport (11.60${\times}$10$^{10}$ mol yr$^{-1}$ Sv$^{-1}$) is about 1.2 larger than that in the Korean Strait (9.72${\times}$10$^{10}$ mol yr$^{-1}$ Sv$^{-1}$) and 2/3 of that by Kuroshio in the East China Sea (18.55${\times}$10$^{10}$ ton yr$^{-1}$ Sv$^{-1}$). It suggests that chemical rich Cheju Current will play a significant role in the biogeochemical processes in the South Sea where the huge land-based waste are introduced.
Purpose : Human umbilical vein endothelial cells(HUVECs) play an important role in regulating blood flow by releasing vasoactive substances. It has been reported that endothelial impairment and dysfunction might be a primary cause of placental vascular disease, which is manifested clinically as preeclampsia in mother and intrauterine growth restriction in fetus. Furthermore, the frequency of apoptotic changes is increased in umbilical and placental tissues from growth-restricted pregnancies. However, the various mechanisms of umbilical endothelial cell apoptosis have not been broadly proposed. We investigate the effects of amiloride derivatives on apoptotic death of HUVECs and identify their ionic mechanism. Methods : HUVECs were purchased from Clonetics, and cultured on endothelial cell growth medium. MTT assay and flow cytometry were used for assessing cytotoxic effect and confirming the apoptosis. Changes in intracellular ion concentrations were measured with specific fluorescent dyes and fluorescence imaging analysis system. Results : Amiloride derivatives elicited cytotoxic effects on HUVECs with dose-dependent manners and the rank order of potency is HMA($IC_{50}\;11.2{\mu}M$), MIA>EIPA>>amiloride. HMA-induced cytotoxicity is dependent on extra- and intracellular pH, that is, increase extra- and intracellular pH augmented the cytotoxic effects of HMA. HMA dose-dependently reduced intracellular major ions, such as $K^+$ and $Cl^-$. Interestingly, the depletion of intracellular ions induced by HMA was also significantly enhanced at alkaline extracellular pH. Conclusion : Amiloride derivatives induce apoptosis of HUVECs with dose and pH dependent manners. They reduce intracellular $K^+$ and $Cl^-$ concentration, which is also extracellular pH dependent.
To research the characteristics of ion currents induced by inhibitory and excitatory herbs of oriental medicine, we used nystatin-perforated patch clamp technique under voltage clamp condition in periaqueductal gray neuron dissociated from Sprauge-Dawley rat, 10-15 days old. The results are as follows. 1. Ion current induced by $10mg/m{\ell}$ of Bupleuri Radix was inhibited $59.50{\pm}4.29%$ by $10^{-4}M$ bicuculline(p>0.01) but inhibition of $10.75{\pm}4.77%$ by $10^{-4}M$ tubocurarine and $4.75{\pm}4.23%$ by $10^{-4}M$ verapamil had no statistical significance(p>0.05). So ion current induced by Bupleuri Radix revealed only GABA induced $Cl^-$ current, not acetylcholine and $Ca^{2+}$ current. 2. Ion current induced by $20mg/m{\ell}$ of Coptidis Rhizoma was inhibited $47.20{\pm}7.88%$ by $10^{-4}M$ bicuculline(p<0.01) but $3.20{\pm}2.33%$ inhibition by $10^{-4}M$ tubocurarine and $1.00{\pm}1.00%$ inhibition by $10^{-4}M$ verapamil had no significance(p>0.05). So ion current induced by Coptidis Rhizoma revealed only GABA induced $Cl^-$ current, not acetylcholine and $Ca^{2+}$ current. 3. Ion current induced by $20mg/m{\ell}$ of Ecliptae Herba was inhibited $55.00{\pm}4.92%$ by $10^{-4}M$ bicuculline (p<0.01), and also inhibited $15.00{\pm}4.26%$ by $10^{-4}M$ tubocurarine(p<0.05), but inhibition of $6.00{\pm}3.03%$ by $10^{-4}M$ verapamil had no significance(p>0.05). So ion current induced by Ecliptae Herba showed GABA activated $Cl^-$ current and acetylcholine activated cation current, not $Ca^{2+}$ current 4. Ion current induced by $5mg/m{\ell}$ of Liriopis Tuber was inhibited $15.20{\pm}4.57%$ by $10^{-4}M$ bicuculline<0.05) and also inhibited $14.00{\pm}3.00%$ by $10^{-4}M$ tubocurarine(p<0.05), but inhibition of $5.20{\pm}4.80%$ by $10^{-4}M$ verapamil had no significance(p>0.05). So ion current induced by Liriopis Tuber showed GABA. activated $Cl^-$ current and acetylcholine activated cation current, not $Ca^{2+}$ current. 5. Ion current induced by $5mg/m{\ell}$ of Aconiti Tuber was inhibited $97.00{\pm}1.34%$ by $10^{-4}M$ bicuculline(p<0.01), $80.00{\pm}9.83%$ by $10^{-4}M$ tubocurarine(p<0.01), and $24.00{\pm}6.18%$ by $10^{-4}M$ verapamil(p<0.05). So ion current induced by Aconiti Tuber revealed GABA activated $Cl^-$ current and acetylcholine activated cation current and $Ca^{2+}$ current. 6. Ion current induced by $10mg/m{\ell}$ of Zingiberis Rhizoma was inhibited $33.00{\pm}7.43%$ by $10^{-4}$ bicuculline(p<0.05), $10.20{\pm}1.83%$ by $10-^{-4}M$ tubocurarine(p<0.01), and $14.00{\pm}2.16%$ by $10^{-4}M$ verapamil(p<0.01) So ion current induced by Zingiberis Rhizoma revealed GABA activated $Cl^-$ current and acetylcholine activated cation outtent and $Ca^{2+}$ current. 7. Ion current induced by $10mg/m{\ell}$ of Boshniakiae Herba was inhibited $65.00{\pm}13.75%$ by $10^{-4}M$ bicuculline(p<0.05), $38.00{\pm}9.24%$ by $10^{-4}M$ tubocurarine(p<0.05), and $33.25{\pm}7.42%$ by $10^{-4}M$ verapamiHp<0.05). So ion current induced by Bpshniakiae Herba revealed GABA activated $Cl^-$ current and acetylcholine activated cation current and $Ca^{2+}$ current. These results suggest that a point of difference between inhibitory and excitatory herbs is existence of$Ca^{2+}$ current.
Glutamate and aspartate may evoke an increase in membrane permeability to monovalent cations and $Ca^{++}$. However, it is uncertain whether $Ca^{++}$ influx is mediated by voltage dependent $Ca^{++}$ channels or by excitatory amino acid activated channels. In addition, the influences of excitatory amino acids on $Ca^{++}$ uptake by neuronal tissues as well as the responses of their actions to extracellular $Mg^{++}$ concentration are different. $K^{+}$ induced $Ca^{++}$ uptake by synaptosomes was dependent on extracellular $Mg^{++}$ up to 5 mM and at concentration of 10 mM, $Ca^{++}$ influx was rather reduced. In $Na^{+}$ rich media, glutamate-and aspartate-induced $Ca^{++}$ uptake was increased by $Mg^{++}$ in a dose independent manner. However, the response for NMDA was inhibited by $Mg^{++}$ at concentrations above 2 mM. $K^+$-and glutamate-induced $Ca^{++}$ influx s were inhibited by 2,4-dinitrophenol, chlorprom-azine and verapamil but not by tetraethylammonium chloride. Tetrodotoxin effectively inhibited the action of glutamate but did not affect that of $K^+$. The response for MNDA was inhibited by 2, 4-dinitrophenol and tetrodotoxin, slightly inhibited by verapamil, and not affected by tetraethylammonium chloride. In $Na^{++}$ rich medium, depolarizing action of glutamate, aspartate and MNDA on synaptosomes was not demonstrated, whereas these agents stimulated $Ca^{++}$ uptake and caused $Ca^{++}$ influx induced depolarization at mitochondria. On the other hand, the activities of synaptosomal ATPases were not affected by excitatory amino acids at 5 mM. The results suggest that glutamate or NMDA induced $Ca^{++}$ influx at synaptosomes exhibits different responses for extracellular $Mg^{++}$ Ex citatory amino acids induced $Ca^{++}$ influx at synaptosomes may be associated with increased permeability of membrane for $Na^{++}$ and $Ca^{++}$ except $K^{++}$ and membrane depolarization due to increased ionic permeability.
In order to investigate histopathologically the nature of varietal resistance and infection process of the rice bast fungus, Pyricularia oryzae. this experiment was undertaken by using the resistant cultivar Tongil and susceptible cultivars Norin No.6 and Jinheung in 1973 to 1974. 1) It was found that appressorium of the fungus forms not only at 4 hours after incubation under the favorable conditions but also peak of appressorial formation is at 48 hours treatment. Physical stimulus known to be a definite factor for appressorial formation. The optimum temperature range for appressorial infection was at $24^{\circ}C$ to $28^{\circ}C$, and pH was between 4.8 to 8.0 with 6.8 as the optimum. 2) Although percent of appressorial formation on the leaves of resistant Tongil and susceptiible Norin No.6 were only slightly different, there was a remarkable difference between resistant and susceptible cultivars with regard to percent of hyphal infection and index for hyphal extension. Index of hyphal extension was 1.6-2.7 in Tonyil. while in susceptible cultivar was 3.4-6.6. The rate of discoloration of infected cells, a indication of hypersensitivity, was greater in the resistant than in susceptible cultvar. 3) Therefore, it could be concluded that resistance of Tongil cultivar may be attributable to a higher degree of mechanical barriers as well as a higher level of antifungal substance accumulation.
Kim, Sung-Jae;Kim, Hae-Min;Choi, Hoon;Kim, Young-Jun
Journal of Life Science
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v.21
no.7
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pp.1032-1038
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2011
Recently identified Ciona intestinalis voltage sensor-containing phosphatase (Ci-VSP) consists of an ion channel-like transmembrane domain (VSD) and a phosphatase-like domain. Ci-VSP senses the change of membrane potential by its VSD and works as a phosphoinositide phosphatase by its phosphatase domain. In this study, we present the construction of His-tagged phosphatase-like domain of Ci-VSP, its recombinant expression and purification, and its enzymatic activity behavior in order to examine the biochemical behavior of phosphatase domain of Ci-VSP without interference. We found that Ci-VSP(248-576)-His can be eluted with an elution buffer containing 25 mM NaCl and 100 mM imidazole during His-tag purification. In addition, we found the proper measurement condition for kinetics study of Ci-VSP(248-576)-His against p-nitrophenyl phosphate (pNPP). We measured the kinetic constant of Ci-VSP(248-576)-His at $37^{\circ}C$, pH 5.0 or 5.5, under 30 min of reaction time, and less than $2.0\;{\mu}g$ of protein amount. With these conditions, we acquired that Ci-VSP(248-576)-His has $K_m$ of $354{\pm}0.143\;{\mu}M$, $V_{max}$ of $0.0607{\pm}0.0137\;{\mu}mol$/min/mg and $k_{cat}$ of $0.359{\pm}0.009751\;min^{-1}$ for pNPP dephosphorylation. Therefore, we produced a pure form of Ci-VSP(248-576)-His, and this showed a higher activity against pNPP. This purified protein will provide the road to a structural investigation on an interesting protein, Ci-VSP.
Gap junction is an ion channel forming between adjacent cells. It also acts as a membrane channel like sodium or potassium channels in a single cell. The amino acid residues up to the $10^{th}$ position in the amino (N)-terminus of gap junction hemichannel affect gating polarity as well as current-voltage (I-V) relation. While wild-type Cx32 channel shows negative gating polarity and inwardly rectifying I-V relation, T8D channel in which threonine residue at $8^{th}$ position is replaced with negatively charged aspartate residue shows reverse gating polarity and linear I-V relation. It is still unclear whether these changes are resulted from the charge effect or the conformational change of the N-terminus. To clarify this issue, we made a mutant channel harboring cysteine residue at the $8^{th}$ position (T8C) and characterized its biophysical properties using substituted-cysteine accessibility method (SCAM). T8C channel shows negative gating polarity and inwardly rectifying I-V relation as wild-type channel does. This result indicates that the substitution of cysteine residue dose not perturb the original conformation of wild-type channel. To elucidate the charge effect two types of methaenthiosulfonate (MTS) reagents (negatively charged $MTSES^-$ and positively charged $MTSET^+$) were used. When $MTSES^-$ was applied, T8C channel behaved as T8D channel, showing positive gating polarity and linear I-V relation. This result indicates that the addition of a negative charge changes the biophysical properties of T8C channel. However, positively charged $MTSET^+$ maintained the main features of T8C channel as expected. It is likely that the addition of a charge by small MTS reagents does not distort the conformation of the N-terminus. Therefore, the opposite effects of $MTSES^-$ and $MTSETT^+$ on T8C channel suggest that the addition of a charge itself rather than the conformational change of the N-terminus changes gating polarity and I-V relation. Furthermore, the accessibility of MTS reagents to amino acid residues at the $8^{th}$ position supports the idea that the N-terminus of gap junction channel forms or lies in the aqueous pore.
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