• Title/Summary/Keyword: AtPIP2-1

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The Optimization for Functional Expression of Arabidopsis Thaliana AtPIP2-1 in Xenopus laevis Oocyte (Xenopus oocyte에서 애기장대 AtPIP2-1 활성측정을 위한 발현 최적화 조건 규명)

  • Kim, Hyun-Mi;Hwang, Hyun-Sik;Lee, Suk-Chan;Jo, Su-Hyun;Kim, Beom-Gi
    • Journal of Applied Biological Chemistry
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    • v.53 no.4
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    • pp.189-194
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    • 2010
  • We confirmed the hypo-osmotic shock strengths and duration, different type of vectors, and subcelluar localization to identify the optimum analysis condition of plant aquaporin activity in Xenopus ooctye using Arabidopsis thaliana AtPIP2-1 gene. Six minutes and 1/5ND buffer hypoosmotic shock treatment was the best condition to show the maximum swelling of Xenopus oocytes where AtPIP2-1 was expressed using pcDNA3.1 vector. AtPIP2-1 protein was expressed more efficiently in pGEMHE vector which has 5' and 3' UTR (untranslation region) of Xenopus ${\beta}$-GLOBIN gene in multiple cloning site than in pcDNA3.1 vector. Also green fluorescence of GFP fused to AtPIP2-1 was detected onto oocyte plasmamembrane where is the proper subcellular localization target of AtPIP2-1.

The Effects of Pomegranate Extracts on the Growth Inhibition against HepG-2 Liver Cancer Cells and Antioxidant Activities (석류 추출물의 간암세포 성장 억제 및 항산화 활성 효과)

  • Park, Kyong-Tae;Kim, Du-Woon;Sin, Tae-Sun;Shim, Sun-Yup;Kim, Mun-Yong;Chun, Soon-Sil
    • Culinary science and hospitality research
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    • v.15 no.1
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    • pp.120-127
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    • 2009
  • In this study, Pip(Pip) and shell powder(Cortex) extracts, and juice concentrates(Juice-1 and Juice-2) of Pomegranate were screened for their growth inhibition on HepG-2 cells and antioxidant activities on DPPH radical-scavenging activity. All samples with higher concentrations showed the growth inhibition against HepG-2 cells. The growth inhibitions against HepG-2 liver cancer cells at 2,500 ppm were in order of Juice-2(43%), Pip(42%), Cortex(38%) and Juice-1(29%). DPPH radical-scavenging activities were higher with the concentrations of the samples increased. The activities of antioxidant BHT, Pip, Cortex, Juice-1, and Juice-2 at 12.5 ppm were 29.9, 16.2, 60.8, 12.6, and 15.1% respectively and those at 25, 50, 100, and 200 ppm were in order of Cortex(81.9$\sim$85.3%), Pip(33.4$\sim$83.0%), BHT(31.3$\sim$47.8%), Juice-2(15.4$\sim$36.8%) and Juice-1(13.4$\sim$36.1%).

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Electrical Characteristics of the PIP Antifuse for Configuration of the Programmable Logic Circuit (프로그램 가능한 논리 회로 구성을 위한 PIP 앤티퓨즈의 전기적 특성)

  • 김필중;윤중현;김종빈
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.14 no.12
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    • pp.953-958
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    • 2001
  • The antifuse is a semi-permanent memory device like a ROM which shows the open or short state, and a switch device connecting logic blocks selectively in FPGA. In addition, the antifuse has been used as a logic device to troubleshoot defective memory cells arising from SDRAM processing. In this study, we have fabricated ONO antifuses consisted of PIP structure. The antifuse shows a high resistance more than several G Ω in the normal state, and shows a low resistance less than 500 Ω after program. The program resistance variation according to temperature shows the very stable value of $\pm$20 Ω. At this time, its program voltage shows 6.7∼7.2 V and the program is performed within 1 second. Therefore this result shows that the PIP antifuse is a very stable and programmable logic device.

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Conflicting Physiological Characteristics and Aquaporin (JcPIP2) Expression of Jatropha (Jatropha curcas L.) as a Bio-energy Crop under Salt and Drought Stresses (바이오에너지 작물 소재로서 자트로파의 염과 가뭄 스트레스 하에서 상반되는 생리적 특성과 아쿠아포린(JcPIP2)의 발현)

  • Jang, Ha-Young;Lee, Ji-Eun;Jang, Young-Seok;Ahn, Sung-Ju
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.3
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    • pp.183-191
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    • 2011
  • This study was undertaken to collect basic knowledge of Jatropha which is one of bio-energy crops, based on the understanding of physiological and molecular aspects under salt and drought conditions. The treatments were followed as: 100, 200 and 300 mM NaCl for salt stress and 5, 10, 20 and 30% PEG for drought stress for 8 days, respectively. Leaf growth, stomatal conductance, chlorophyll fluorescence and gene expression of aquaporin (JcPIP2) of Jatropha were investigated. From 2 days after treatments, plants treated with higher than 100 mM NaCl and 10% PEG respectively were significantly suppressed in leaf length, width, and stomatal conductance, but 5% PEG treatment showed that plant growth was improved more than control plant. Semi-quantitative RT-PCR analyses revealed that the JcPIP2 gene was expressed in root, stem, cotyledon and leaves. It was not detected in leaves at 200 and 300 mM NaCl treatments. However, transcripts of JcPIP2 were induced in roots and stems under salt and drought conditions compared to those of healthy plants. Therefore, it was concluded that JcPIP2 plays an important role in improving drought tolerance.

Regulation of Adenosine-activated GIRK Channels by Gq-coupled Receptors in Mouse Atrial Myocytes

  • Cho, Ha-Na
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.3
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    • pp.145-150
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    • 2010
  • Adenosine (Ado) is an important mediator of the endogenous defense against ischemia-induced injury in the heart. The action of Ado is mediated by activation of G protein-gated inwardly rectifying $K^+$ (GIRK) channels. In turn, GIRK channels are inhibited by reducing phosphatidylinositol 4,5-bisphosphate ($PIP_2$) through Gq protein-coupled receptors (GqPCRs). We previously found that GIRK channels activated by acetylcholine, a muscarinic M2 acetylcholine receptor agonist, are inhibited by GqPCRs in a receptor-specific manner. However, it is not known whether GIRK channels activated by Ado signaling are also regulated by GqPCRs. Presently, this was investigated in mouse atrial myocytes using the patch clamp technique. GIRK channels were activated by $100\;{\mu}M$ Ado. When Ado was repetitively applied at intervals of 5~6 min, the amplitude of second Ado-activated GIRK currents ($I_{K(Ado)}$) was $88.3{\pm}3.7%$ of the first $I_{K(Ado)}$ in the control. Pretreatment of atrial myocytes with phenylephrine, endothelin-1, or bradykinin prior to a second application of Ado reduced the amplitude of the second $I_{K(Ado)}$ to $25.5{\pm}11.6%$, $30.5{\pm}5.6%$, and $96.0{\pm}2.7%$, respectively. The potency of $I_{K(Ado)}$ inhibition by GqPCRs was different with that observed in acetylcholine-activated GIRK currents ($I_{K(ACh)}$) (endothelin-1>phenylephrine>bradykinin). $I_{K(Ado)}$ was almost completely inhibited by $500\;{\mu}M$ of the $PIP_2$ scavenger neomycin, suggesting low $PIP_2$ affinity of $I_{K(Ado)}$. Taken together, these results suggest that the crosstalk between GqPCRs and the Ado-induced signaling pathway is receptor-specific. The differential change in $PIP_2$ affinity of GIRK channels activated by Ado and ACh may underlie, at least in part, their differential responses to GqPCR agonists.

MMP-1 and PIP Expressions from Ethanol Extract of Hydnocarpus anthelmintica Pierre in Human Fibroblast Cells (사람유래 섬유아세포에서 대풍자 에탄올 추출물의 MMP-1과 PIP의 발현에 대한 연구)

  • Choi, Eun-Young;Jang, Young-Ah;Ki, Se-Gie
    • Journal of Life Science
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    • v.32 no.12
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    • pp.938-946
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    • 2022
  • This study aims to evaluate the effects of antioxidant activities, protein and mRNA expressions of matrix metalloproteinase (MMP) -1 and procollagen type I C-peptide (PIP) in 70% ethanol extract from Hydnocarpus anthelmintica Pierre (HE). DPPH and ABTS+ radicals scavenging assays were measured for antioxidant activities and HE had 73.5% and 74.4% of scavenging activities at 1,000 ㎍/ml concentration, respectively. And we investigated the inhibition of collagenase by HE, and the result was a 78.8% inhibition effect on concentrations of 1,000 ㎍/ml. In addition, an MTT assay was performed to confirm the toxicity of the CCD-986sk fibroblasts to the HE, and as a result, the cell viability rate was about 91.7% at a concentration of 50 ㎍/ml or less, and subsequent cell experiments were performed at a concentration of 50 ㎍/ml or less. We treated the cells with UVB (20 mJ/cm2) for stimulation, treated HE at various concentrations, and performed ELISA tests and RT-PCR experiments. And HE increased the PIP and mRNA in a dose-dependent manner and showed an expression rate of about 64.2% and 83.4%, respectively, at a concentration of 50 ㎍/ml compared with Cont (50.3% and 45.8%, respectively). And HE suppressed the MMP-1 protein and mRNA in a dose-dependent manner and showed a low expression rate of about 48.7% and 35.9%, respectively, at a concentration of 50 ㎍/ml. These results can be applied to developing anti-wrinkle materials for functional food and cosmetics with HE.

Expression of CsRCI2s by NaCl stress reduces water and sodium ion permeation through CsPIP2;1 in Camelina sativa L.

  • Kim, Hyun-Sung;Lim, Hyun-Gyu;Ahn, Sung-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.194-194
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    • 2017
  • Camelina (Camelina sativa L.) is a potential bio-energy crop that has short life cycle about 90 days and contains high amount of unsaturated fatty acid which is adequate to bio-diesel production. Enhancing environmental stress tolerance is a main issue to increase not only crop productivity but also big mass production. CsRCI2s (Rare Cold Inducible 2) are cold and salt stress related protein that localized at plasma membrane (PM) and assume to be membrane potential regulation factor. These proteins can be divide into C-terminal tail (CsRCI2D/E/F/G) or no-tail group (CsRCI2A/B/C/H). However, function of CsRCI2s are less understood. In this study, physiological responses and functional characterization of CsRCI2s of Camelina under salt stress were analyzed. Full-length CsRCI2s (A/B/E/F) and CsPIP2;1 sequences were confirmed from Camelina genome browser. Physiological investigations were carried out using one- or four-week-old Camelina under NaCl stress with dose and time dependent manner. Transcriptional changes of CsRCI2A/B/E/F and CsPIP2;1 were determined using qRT-PCR in one-week-old Camelina seedlings treated with NaCl. Translational changes of CsRCI2E and CsPIP2;1 were confirmed with western-blot using the antibodies. Water transport activity and membrane potential measurement were observed by cRNA injected Xenopus laevis oocyte. As results, root growth rate and physiological parameters such as stomatal conductance, chlorophyll fluorescence, and electrolyte leakage showed significant inhibition in 100 and 150 mM NaCl. Transcriptional level of CsPIP2;1 did not changed but CsRCI2s were significantly increased by NaCl concentration, however, no-tail type CsRCI2A and CsRCI2B increased earlier than tail type CsRCI2E and CsRCI2F. Translational changes of CsPIP2;1 was constitutively maintained under NaCl stress. But, accumulation of CsRCI2E significantly increased by NaCl stress. CsPIP2;1 and CsRCI2A/B/E/F co-expressed Xenopus laevis oocyte showed decreased water transport activity as 61.84, 60.30, 62.91 and 76.51 % at CsRCI2A, CsRCI2B, CsRCI2E and CsRCI2F co-expression when compare with single expression of CsPIP2;1, respectively. Moreover, oocyte membrane potential was significantly hyperpolarized by co-expression of CsRCI2s. However, higher hyperpolarized level was observed in tail-type CsRCI2E and CsRCI2F than others, especially, CsRCI2E showed highest level. It means transport of $Na^+$ ion into cell is negatively regulated by expression of CsRCI2s, and, function of C-terminal tail is might be related with $Na^+$ ion influx. In conclusion, accumulation of NaCl-induced CsRCI2 proteins are related with $Na^+$ ion exclusion and prevent water loss by CsPIP2;1 under NaCl stress.

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Water relations of plants under environmental stresses: role of aquaporins

  • Kang, H.S.;Ahn, S.J.;Hong, S.W.;Chung, G.C.
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.71-80
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    • 2005
  • Effects of low temperature ($8^{\circ}C$) on the hydraulic conductivity of young roots of a chilling-sensitive (cucumber; Cucumis sativus L.) and a chilling-resistant (figleaf gourd; Cucurbita ficifolia Bouche) crop have been measured at the levels of whole root systems (root hydraulic conductivity, $Lp_r$) and of individual cortical cells (cell hydraulic conductivity, Lp). In figleaf gourd, there was a reduction only in hydrostatic $Lp_r$ but not in osmotic $Lp_r$ suggesting that the activity of water channels was not much affected by low root temperature (LRT)treatment in this species. Changes in cell Lp in response to chilling and recovery were similar asroot level, although they were more intense at the root level. Roots of figleaf gourd recovered better from LRT treatment than those of cucumber. In figleaf gourd, recovery (both at the root and cell level) often resulted in Lp and $Lp_r$ values which were even bigger than the original, i.e. there was an overshoot in hydraulic conductivity. These effects were larger forosmotic (representing the cell-to-cell passage of water) than for hydrostatic $Lp_r$. After a short term (1 d) exposure to $8\;^{\circ}C$ followed by 1 d at $20\;^{\circ}C$, hydrostatic $Lp_r$ of cucumber nearly recovered and that of figleaf gourd still remained higher due to the overshoot. On the contrary, osmotic $Lp_r$ and cell Lp in both species remained high by a factor of 3 as compared to the control, possibly due to an increased activity of water channels. After pre-conditioning of roots at LRT, increased hydraulic conductivitywas completely inhibited by $HgCl_2$ at both the root and cell levels. Different from figleaf gourd, recovery from chilling was not complete in cucumber after longer exposure to LRT. It is concluded that at LRT, both changes in the activity of aquaporins and alterations of root anatomy determine the water uptake in both species. To better understand the aquaporin function in plants under various stress conditions, we examined the transgenic Arabidopsisand tobacco plants that constitutively overexpress ArabidopsisPIP1;4 or PIP2;5 under various abiotic stress conditions. No significant differences in growth rates were found between the transgenic and wild-type plants under favorable growth conditions. By contrast, overexpression of PIP1;4 or PIP2;5 had a negative effect on seed germination and seedling growth under drought stress, whereas it had a positive effect under cold stress and no effect under salt stress. Measurement of water transport by cell pressure probe revealed that these observed phenotypes under different stress conditions were closely correlated with the ability of water transport by each aquaporin in the transgenic plants. Together, our results demonstrate that PIP-type aquaporins play roles in seed germination, seedling growth, and stress response of Arabidopsis and tobacco plants under various stress conditions, and emphasize the importance of a single aquaporin-mediated water transport in these cellular processes.

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N1-N2 Audiograms of the Guinea Pig Cochlea (기네아피그 달팽이관의 N1-N2 오디오그램)

  • 장순석
    • Journal of Biomedical Engineering Research
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    • v.16 no.1
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    • pp.77-84
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    • 1995
  • Nl and N2 gross neural action potentials were measured from the round window of the guinea pig cochlea at the onset of the acoustic stimuli. Nl -N2 audiograms were made by means of regulating stimulant intensities in order to produce constant Nl -N2 potentials as criteria for different input tone pip frequencies. The lowest threshold was measured with an input tone pip 15 dB SPL in intensity and 12 KHz in frequency when the animal was in normal physiological condition. The procedure of experimental measurements is explained in detail. This experimental approach is very useful for the investigation of the Cochlear function. Both nonlinear and active functions of the Cochlea can be monitored by Nl -N2 audiograms. Key words : Guinea Pig, Cochlea, Wl and N2 Gross Neural Action Potential, Nl -N2 audiogram.

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Preparation of Pore-filled Ion-exchange Membranes using Poly(vinylbenzyl ammoninum salt) (Poly(vinylbenzyl ammonium salt)를 이용한 Pore-filled 이온교환막의 제조)

  • 변홍식
    • Membrane Journal
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    • v.11 no.3
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    • pp.109-115
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    • 2001
  • Pore-filled ion-exchange membranes in which polypropylene(PP) microporous membrane was used as a nascent membrane were prepared by an in-situ cross-linking technique. Poly(vinylbenzyl chloride)(PVBCI) reacted with piperazine(PIP) or 1,4-diaminobicyclo[2,2,2]octane(DABCO) in a di-methylforamide(DMF) solution was filled in the pores of the microporous base membrane. After gellation the remaining chloromethyl groups were, then reacted with an amine such as trimethylamine to form positively charged, ammonium site. This will produce the pore-filled anion-exchange membrane. It was shown that this simple 2 step procedure gave dimensionally stable, pore-filled membranes in which the MG of polymer gel and degree of cross-linking could be easily controlled by the concentration of PVBCI and cross-linker in the starting DMF solution. Specially, high water permeability (7.8 kg/$m^2$hr, host membrane: PP3, MG: 73%, degree of cross-linking: 10%, crosslinker: PIP) at ultra low pressure(100 kPa) indicates the produced pore-filled membranes is usable as a water softening membrane.

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