• Animal cells and systems
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• 제16권5호
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• pp.376-384
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• 2012

Although ginsenosides have a variety of physiologic or pharmacologic functions in various regions, there are only a few reports on the effects of transient receptor potential melastatin 7 (TRPM7) channels. Here, we showed evidence suggesting that TRPM7 channels play an important role in ginseng total saponin (GTS)-mediated cellular injury. The combination techniques of electrophysiology, pharmacological analysis, small interfering RNA (siRNA) method and cell death assays were used. GTS depolarized the resting membrane potentials and decreased the amplitude of pacemaker potentials in cultured interstitial cells of Cajal (ICCs) in gastrointestinal (GI) tract. The TRPM7-like currents in single ICCs and the overexpressing TRPM7 in HEK293 cells were inhibited by GTS. However, GTS had no effect on $Ca^{2+}$-activated $Cl^-$ conductance. GTS inhibited the survival of human gastric (AGS) and brea (MCF-7) adenocarcinoma cells. Also, GTS inhibited the TRPM7-like currents in AGS and MCF-7 cells. The GTS-mediated cytotoxicity was inhibited by TRPM7-specific siRNA. In addition, we showed that overexpression of TRPM7 channels in HEK293 cells was inhibited by GTS. Thus, TRPM7 channels are involved in GTS-mediated cell death in AGS and MCF-7 cells, and these channels may represent a novel target for physiological disorders where GTS plays an important role.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.677-684
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• 2000

Pseudomonas sp. BK7, an alkalophile, displayed the highest growth and protease activity when grown in a fermenter which was controlled at a pH level of 9.0, and the enzyme production was significantly enhanced by the increase of agitation speed. Two forms of alkaline proteases (BK7-1 and BK7-2) were fractionated and purified to near homogeneity. Protease BK7-1 was purified through CM-Sepharose CL-6B and Sephadex G-75 column chromatographies, and Protease BK7-2 was purified through CM-Sepharose CL-6B, DEAE-Sepharose, and Sephadex G-75 column chromatographies. The molecular weights of proteases BK7-1 and BK7-2 determined by gel filtration chromatography were 20,700 and 40,800, respectively. The $K_m$ value, isoelectric point, and optimum pH of protease BK7-1 were 2.55 mg/ml, 11.0, and 11.0, respectively, whereas those of protease BK7-2 were 1.57 mg/ml, 7.2, and 10.0, respectively. Both proteases were practically stable in the pH range of 5-11. The optimum temperatures for the activities of both protease BK7-1 and BK7-2 were $50^{\circ}C$ and $45^{\circ}C$, respectively. About 56% of the original protease BK7-2 activity remained after being treated at $50^{\circ}C$ for 30 min but protease BK7-1 was rapidly inactivated at above $25^{\circ}C$. Both proteases were completely inhibited by phenylmethane sulfonyl fluoride, a serine protease inhibitor. Protease BK7-2 was stable against EDTA, EGTA, STP, and detergents such as SDS and LAS, whereas protease BK7-1 was found to be unstable.

• Journal of Ginseng Research
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• 제37권2호
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• pp.201-209
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• 2013

Ginsenoside, one of the active ingredients of Panax ginseng, has a variety of physiologic and pharmacologic effects. The purpose of this study was to explore the effects of ginsenoside Rd (G-Rd) on melastatin type transient receptor potential 7 (TRPM7) channels with respect to the proliferation and survival of AGS and MCF-7 cells (a gastric and a breast cancer cell line, respectively). AGS and MCF-7 cells were treated with different concentrations of G-Rd, and caspase-3 activities, mitochondrial depolarizations, and sub-G1 fractions were analyzed to determine if cell death occurred by apoptosis. In addition, human embryonic kidney (HEK) 293 cells overexpressing TRPM7 channels were used to confirm the role of TRPM7 channels. G-Rd inhibited the proliferation and survival of AGS and MCF-7 cells and enhanced caspase-3 activity, mitochondrial depolarization, and sub-G1 populations. In addition, G-Rd inhibited TRPM7-like currents in AGS and MCF-7 cells and in TRPM7 channel overexpressing HEK 293 cells, as determined by whole cell voltage-clamp recordings. Furthermore, TRPM7 overexpression in HEK 293 cells promoted G-Rd induced cell death. These findings suggest that G-Rd inhibits the proliferation and survival of gastric and breast cancer cells by inhibiting TRPM7 channel activity.

• BMB Reports
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• 제54권6호
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• pp.311-316
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• 2021

Ethanol often causes critical health problems by altering the neuronal activities of the central and peripheral nerve systems. One of the cellular targets of ethanol is the plasma membrane proteins including ion channels and receptors. Recently, we reported that ethanol elevates membrane excitability in sympathetic neurons by inhibiting Kv7.2/7.3 channels in a cell type-specific manner. Even though our studies revealed that the inhibitory effects of ethanol on the Kv7.2/7.3 channel was diminished by the increase of plasma membrane phosphatidylinositol 4,5-bisphosphate (PI(4,5)P₂), the molecular mechanism of ethanol on Kv7.2/7.3 channel inhibition remains unclear. By investigating the kinetics of Kv7.2/7.3 current in high K⁺ solution, we found that ethanol inhibited Kv7.2/7.3 channels through a mechanism distinct from that of tetraethylammonium (TEA) which enters into the pore and blocks the gate of the channels. Using a non-stationary noise analysis (NSNA), we demonstrated that the inhibitory effect of ethanol is the result of reduction of open probability (P_O) of the Kv7.2/7.3 channel, but not of a single channel current (i) or channel number (N). Finally, ethanol selectively facilitated the kinetics of Kv7.2 current suppression by voltage-sensing phosphatase (VSP)-induced PI(4,5)P₂ depletion, while it slowed down Kv7.2 current recovery from the VSP-induced inhibition. Together our results suggest that ethanol regulates neuronal activity through the reduction of open probability and PI(4,5)P₂ sensitivity of Kv7.2/7.3 channels.

• Applied Biological Chemistry
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• 제20권1호
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• pp.26-32
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• 1977

대두(大豆)(Glycine max cultivar Gwang-gyo)의 각 성숙시기(成熟時期)에 나타나는 7S globulin을 분리하여 Davis 방법(方法)에 의한 전기영동과 PAWU용매에 의해서 유리(遊離)되는 그들의 subunit를 전기영동한 결과 7S globulin중의 복합단백질간(複合蛋白質間)에는 그 구성(構成) subunit에 유사성(類似性)이 있음을 시사하였다. 7S globulin의 복합단백질(複合蛋白質)을 DEAE-Sephadex A-50으로 크로마토그라피하여 분리하였다. 이때 pH 7.6의 인산완충액(燐酸緩衝液)에서 NaCl의 농도구배(濃度句配)가 0.28M부터 0.40M 사이에서 두 개의 분획(分劃)으로 분리되었다. 이들 명(名) 단백질(蛋白質)의 subunit를 5M urea와 1% SDS로 유리(遊離)시켜 7.5% acrylamide-PAWU gel과 5.6% acrylamide-SDS gel에서 전기영동하였다. 그 결과 subunit의 하전량(荷電量)에 의해서 분리되는 PAWU gel전기영동에서 7S globulin이 5개의 주 분리대로 분리되고 그중 2개의 분리대가 7S-A globulin과 7S-B globulin에 공유(共有)되어 있었다. 또 subunit의 분자량(分子量)에 따라서 분리되는 SDS gel 전기영동에서는 7S globulin이 7개의 주 분리대를 나타내는데 그 중에서 3개의 분리대가 7S-A와 7S-B 분획에 공유(共有)되어 있었다. 따라서 7S globulin의 복합단백질간(複合蛋白質間)에는 구성(構成) subunit간(間)에 유사성(類似性)이 있는 것으로 나타났다.

• 한국생물환경조절학회:학술대회논문집
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• 한국생물환경조절학회 2001년도 봄 학술발표논문집
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• pp.51-56
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• 2001

This study was carried out to research the effect of nutrient composition and GA4＋7 on growth, flower quality and bulb development in special hydroponics of cut tulip. Flowering of 'Cassini' was markedly accelerated by standard+GA4＋7, K200＋GA4＋7, N250＋GA4＋7 treatment, last internode was increased by N250 as compared with other treatment. For 'Christmas Marvel', flowering was promoted by K250＋GA4＋7, last infernode and total length were also increased with K250＋GA4＋7 as compared to other treatment. For 'Golden Apeldoorn', flowering was accelerated by K200＋GA4＋7, last internode was increased with N300＋GA4＋7 treatment and total length was increased in N250＋GA4＋7 treatment as compared with the standard plants. Bulb weight and number of 'Cassini', 'Christmas Marvel'and 'Golden Apeldoorn' were decreased by GA4＋7 as compared to non-treatment, and bulb diameter was not affected by GA4＋7.

• 교육녹색환경연구
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• 제17권1호
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• pp.33-39
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• 2018

초등학교 학급당 학생수가 점차로 줄어 이제는 OECD 평균 21.1명(2017년 기준) 인구추계를 따르게 되면 20.5명(2025년)을 거쳐 19.8명(2030년)이 된다고 한다. 이러한 추계로 보았을 때 초등학교 학급당 학생수는 20~22명 정도가 향후 오랜 기간 동안 유지될 가능성이 높다. 현재의 교실들의 크기는 최소 30명을 수용할 수 있는 구조여서 많으면 10명 정도 더 과잉된 교실면적이 할애되고 있어 예산의 낭비와 공간의 밀도가 떨어지고 있다. 이에 본 연구는 학생 1인당 단위공간을 기초로 하여 일제식 수업 및 모둠수업에 필요한 공간을 도면으로 분석한 결과 다음과 같은 결론을 얻었다. 2인 1조의 일제식 수업에 필요한 교실크기는 벽체간 두께를 200으로 하였을 때 가로의 중심선간 치수는 7,100~7,500이 되고 세로는 7,000~7,800이 된다. 이러한 치수에 대하여 30cm 모듈에 맞게 적용하면 가로는 7.2m 7.5m, 세로 7.2m, 7.5m, 7.8m이 된다. 따라서 단위교실의 모듈은 정방형일 경우 $7.2m{\times}7.2m$, $7.5m{\times}7.5m$, 장방형일 경우 $7.2m{\times}7.5m$, $7.2m{\times}7.8m$, $7.5m{\times}7.8m$의 모듈이 나올 수 있다. 모듈의 면적은 $7.2m{\times}7.2m(51.84m^2)$, $7.5m{\times}7.5m(56.25m^2)$, $7.2m{\times}7.5m(54m^2)$, $7.2m{\times}7.8m(56.16m^2)$, $7.5m{\times}7.8m(58.5m^2)$로서 최소 $51.84m^2$에서 최대 $58.5m^2$가 된다. 이러한 면적은 기존의 교실 모듈 $7.5m{\times}9.0m(67.5m^2)$, $8.0m{\times}8.0m(64m^2)$, $8.1m{\times}8.1m(65.61m^2)$, $8.4m{\times}8.0m(67.2m^2)$ 등과 비교하면 최소 $5.5m^2$에서 최대 $15.7m^2$의 면적을 줄일 수 있다.

• 한국발생생물학회지:발생과생식
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• 제26권3호
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• pp.117-126
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• 2022

Bromodomain-containing protein 7 (BRD7) participates in many cellular processes and embryo development. BRD7 is down-regulated in various cancers and evidence of its tumor suppressor function has been accumulating. Here, we identified transforming stimulated clone 22 (TSC-22) as a novel BRD7 interacting protein and show its novel function as a positive regulator of BRD7. We found that TSC-22 expression potentiated the inactivation of the extracellular signal-regulate kinase (ERK) pathway by BRD7. Our data establishes TSC-22 as a modulator of BRD7 and unravels the molecular mechanisms that drive the synergistic tumor-suppressing effects of TSC-22 and BRD7. Our findings may open new avenues for developing novel molecular therapies for tumors exhibiting down-regulated BRD7 and/or TSC-22.

• Journal of Microbiology and Biotechnology
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• 제6권5호
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• pp.336-339
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• 1996

Glutaryl 7-aminocephalosporanic acid acylase of Pseudomonas sp. SY-77-1 was immobilized with oxiran acrylic beads for the production of 7-aminocephalosporanic acid (7-ACA) from glutaryl 7-aminocephalosporanic acid (GL 7-ACA). The immobilized enzyme maintained its activity at a constant level for 7 days, but lost 30$%$ of its activity after 20 days. Optimal reaction conditions for the synthesis of 7-ACA were found to be $30^{\circ}C$ and pH 8.0 using the immobilized enzyme. For the economic production of 7-ACA, substrate and enzyme concentrations were optimized to 60 mM and 0.5 g wet weight per 10 $m\ell$ of reaction volume, respectively. Under optimized conditions, 50 mM 7-ACA was produced from 60mM GL 7-ACA within 8 h, resulting in a conversion yield of 83$%$.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.452-457
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• 2001

The glutaryl 7-aminocephalosporanic acid (glutaryl 7-ACA) acylase was purified from Pseudomonas diminuta KAC-1 cells isolated from soil, and characterized. The acylase was purified by procedures including ammonium sulfate fractionation and column chromatographies on DEAE-Sepharose, Phenyl-Sepharose, Q-Sepharose, and Superose 12H/R. The negative acylase was found to be composed of two subunits with molecular masses of approximately 55 kDa and 17 kDa, respectively. The isoelectric point of the enzyme was 4.0. The specific activities of the purified acylase were 8.0 and 7.0 U/mg on glutaryl 7-ACA and glutaryl 7-aminodesacetoxy cephalosporanic acid (glutaryl 7-ADCA), respectively, and $K_m$ values were 0.45 mM for glutaryl 7-ADCA and 0.67 mM for glutaryl 7-ADCA. The enzyme had a pH optimum at 8.0 and a tmperature optimum at $40^{\circ}C$. The acylase catalyzed the synthesis of glutaryl 7-ACA from glutaric acid and 7-ACA as well as the hydrolysis of glutaryl 7-ADCA, although the reaction rate of the synthesis was slower than that of the hydrolysis. In addition, it was found that the enzyme had a glutaryl transferase activity, thereby transferring the glutaryl group from one cephalosporin nucleus to another.