• Title/Summary/Keyword: GluR1

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The Role of Aquaporin-4 in Cerebral Edema Formation after Focal Cerebral Ischemia in Rats

  • Song, Young-Jin;Bae, Hae-Rahn;Ha, Se-Un;Huh, Jae-Taeck
    • Journal of Korean Neurosurgical Society
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    • v.41 no.1
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    • pp.30-38
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    • 2007
  • Objective : To elucidate the role of aquaporin-4[AQP4] in cerebral edema formation, we studied the expression and subcellular localization of AQP4 in astrocytes after focal cerebral ischemia. Methods : Cerebral ischemia were induced by permanent middle cerebral artery[MCA] occlusion in rats and estimated by the discoloration after triphenyltetrazolium chloride[TTC] immersion. Change of AQP4 expression were evaluated using western blot. Localization of AQP4 was assessed by confocal microscopy and its interaction with ${\alpha}-syntrophin$ was analyzed by immunoprecipitation. Results : After right MCA occlusion, the size of infarct and number of apoptotic cells increased with time. The ratio of GluR1/GluR2 expression also increased during ischemia. The polarized localization of AQP4 in the endfeet of astrocytes contacting with ventricles, vessels and pia mater was changed into the diffuse distribution in cytoplasm. The interactions of AQP4 and Kir with ${\alpha}-syntrophin$, an adaptor of dystrophin complex, were disrupted by cerebral ischemia. Conclusion : The deranged spatial buffering function of astrocytes due to mislocalized AQP4/Kir4.1 channel as well as increased assembly of $Ca^{2+}$ permeable AMPA receptors might contribute to the development of edema formation and the excitotoxic neuronal cell death during ischemia.

Structure and $Ca^{2+}$-ion effects on the function of $\alpha$-cyclodextrin Glucanotransferase from B. macerans : An X-ray study (Bacillus macerans에서 정제한 $\alpha$-cyclooextrin glucanotransferase의 구조와 칼슘이온이 기능에 미치는 영향 : X-ray 연구)

  • 최희욱;홍순강
    • KSBB Journal
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    • v.19 no.2
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    • pp.159-163
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    • 2004
  • The X-ray structure of the cydodextrin-glucanotransferase of Bacillus macerans was solved by molecular replacement at 2.0 ${\AA}$ resolution. The refined structure has a crystallographic R-factor of 16.6%, (R$\sub$free/ = 20.5%). A new metal binding site occupied by two Ca$\^$2+/-ions was found at an accession channel of the active site. There is a large accumulation of negative charges that bind these Ca$\^$2+/-ions, thereby connecting segment ${\beta}$13-${\alpha}$G (residue 254-276) to the main body of domain A (at ${\alpha}$H, residue 283-297). The segment 313-${\alpha}$G contains the catalytic residue Glu258 between subsite 1 and -1 and Tyr260 (subsite 2) which is located at the entrance of the active site. The Ca$\^$2+/-site 3a,b may have a major role for the activity and specificity of this CGTase, although it is not even conserved for the a-subclass of CGTases.

Isolation and Characterization of Gelidibacter sp. HK-1 Producing Alkaline Protease (알칼리성 단백질 분해 효소 생산 균주 Gelidibacter sp. HK-1의 분리 및 특성)

  • Oh, Hyun-Geun;Lee, Soon-Youl;Lee, Jae-Hag
    • The Korean Journal of Food And Nutrition
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    • v.19 no.4
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    • pp.496-501
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    • 2006
  • This study was to isolate a bacterium producing a alkaline protease from mud flats of the west seaside of Korea and to investigate the biochemical analysis of the alkaline protease producing from the isolate. The isolate was named as Gelidibacter sp. HK-1 based on 16S rRNA sequence, Gram staining and the photograph of electron microsceope. Optimum temperature for growth and pretense production of the isolate was $25^{\circ}C$. Growth of the isolate was reached at stationary phase after 10hrs followed by inoculation. Maximum activity of protease produced from the isolate was shown after 14hrs. Optimum temperature and pH for the protease activity were $45^{\circ}C$ and pH 9, respectively. Molecular weight of the pretense was about 50KD and the partial amino acid sequence of the pretense was Ala-Try-Ala-Leu-Asn-Thr-Ser-Val-Thr-Glu-Thr-Phe-Ala-Lys. The partial amino acid sequences of the protease showed significant homology with a pretense produced from Streptomyces avermitilis.

Use of 16S-23S rRNA Intergenic Spacer Region for Rapid Detection of Vibrio fluvialis (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio fluvialis의 검출)

  • 강현실;허문수;이제희
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.77-85
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    • 2003
  • We have examined the 16S-23S rRNA intergenic spacer regions (ISR) of Vibrio fluvialis. ISRs were PCR amplified, cloned into a plasmid vector and then sequenced. As results of ISR nucleotide sequence analysis, total of 6 clones were isolated depending on the size. The clones were different in both the number and the composition of the tRNA genes, and were designated ISR-A, ISR-E, ISR-El, ISR-lA, ISR-EKV, ISR-EKAV. ISR-A contains $tRNA^{Ala}$; ISR-lA, $tRNA^{Ile}$-$tRNA^{Ala}$; ISR-EKV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Val}$;ISE-EKAV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Ala}$-$tRNA^{Val}$; ISR -E and E1, $tRNA^{GIu}$ clusters. ISR-EKV was shown to be a minor type out of the six ISR types and showed a very limited homology between ISR-EKV from V, fluvialis and ISRa from other Vibrio species. Therefore ISR-EKV sequence was used to design species-specific primers to detect V, fiuvialis from other Vibrio species by PCR reaction. The specificity of the primers was examined using genomic DNA of other Vibrios as templates for PCR reaction. The result showed that PCR can be a useful method to detect V. fluvialis among Vibrio species in a single PCR reaction.

Moisturizing Effects and Composition Analysis of Proteoglycan Isolated from Chia (Salvia hispanica) Seed (치아 씨앗으로부터 분리한 단백다당체의 성분분석과 보습 효과)

  • Lee, Bum-Chun;Joo, Chul-Gue;Hur, Ji-Yeon;Lee, Keun-Ha;Kim, Young-Jin;Lee, Chan-Woo;Kim, Jin-Woong;Park, Yong-Il;Kim, Hyun-Sook;Choe, Tae-Boo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.2
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    • pp.121-128
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    • 2010
  • We investigated the effects of skin hydration and composition analysis of proteoglycan (chia seed polymer) produced from chia (Salvia hispanica) seed. The result showed that proteoglycan of chia seeds is composed of galactose (46.8 %), glucuronic acid (27.1 %), rhamnose (8.7 %), xylose (7.6 %), glucose (4.9 %), fructose (2.3 %), mannose (1.8 %), arabinose (0.9 %) and the amount of proteins contained is 31.3 mg/g with the constituent amino acid compositions (mg/g) of Asp (1.9), Glu (3.6), Ser (0.9), Gly (3.6), Thr (0.8), Arg (1.0), Ala (2.0), Tyr (0.4), Cys (4.8), Val (1.1), Phe (0.5), Ile (0.6), Leu (0.9). The molecular weight of the proteoglycan measured by GPC (Gel Permeation Chromatography) is the range of 100,000~250,000 Da and the average molecular weight is 170,000 Da. The moisturizing effects and trans-epidermal water loss (TEWL) of chia seed polymer in cosmetic products (O/W emulsion) were studied in vivo. Chia seed polymer showed good skin hydration effects when compared with sodium hyaluronate which is a common moisturizer. Taken all together, chia seed polymer should be a very useful cosmetic ingredient as a moisturizer and a protecting agent from various skin irritations.

Effects of acupuncture at the $LU8{\cdot}K17$ on Anti-apoptotic cell death and neuroprotection in Rat hippocampus following focal brain ischemic injury induced by Intraluminal Filament insertion in Rats (직자(直刺), 영수(迎隨) 및 염전수기법(捻轉手技法)에 따라 시행한 경거(經渠)${\cdot}$복류(復溜) 침자(鍼刺)가 중대뇌동맥(中大腦動脈) 폐새(閉塞)에 의하여 유발(誘發)된 국소(局所) 뇌허혈(腦虛血) 백서(白鼠) hippocampus의 항세포자멸사(抗細胞自滅死) 및 복경보호(福經保護)에 미치는 영향(影響))

  • Youn, Dae-Hwan;Byun, Jeng-Yun;Choi, Chan-Hun;Baek, Jin-Ung;Jeong, Ji-Yeon;Jung, Yeon-Jin;Na, Chang-Su
    • Korean Journal of Acupuncture
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    • v.24 no.3
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    • pp.205-221
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    • 2007
  • Objectives : Aims of this study is to investigate the effects of $LU8{\cdot}KI7$ in rat induced by experimental focal ischemia. Materials and methods : The focal ischemia was induced by intraluminal filament insertion into middle cerebral artery. The groups divided into 6groups, control; no therapy group after ischemia-induced, AT1; acupuncture therapy group at $LU8{\cdot}KI7$ after ischemia-induced, AT2; acupuncture therapy at $LU8{\cdot}KI7$ bilaterally and the needle was twirled and rotated forward with the thumb of the right hand 9times, AT3; acupuncture therapy at $LU8{\cdot}KI7$ bilaterally and the needle was twirled and rotated forward with the forefinger of the right hand 9times, AT4; acupuncture therapy at$LU8{\cdot}KI7$ bilaterally and the needle was inserted to the direction following the flowing route of the meridian(digital direction), AT5; acupuncture therapy at $LU8{\cdot}KI7$ bilaterally, the needle was inserted to the direction following the flowing route of the meridian(digital direction) and the needle was twirled and rotated forward with the thumb of the right hand 9times. Acupuncture therapy was carried out 7times during 2weeks after focal ischemia-induced. The anti-apoptotic and neuroprotective effects of acupuncture are observed by Bax, Bcl-2, mGluR5, cytochrome c, Cresyl violet and ChAT-stain. Results : The intensity of Bax was decreased in AC1, AC4, AC5 group, was increased in AC2, AC3 group. The intensity of Bcl-2 was increased in AC2, AC3, AC4, AC5 group. The intensity of mGluR5 was decreased in AC1 group, was increased in AC4, AC5 group. The intensity of Cytochrome c was increased in ACI, AC2 group, was decreased in AC4, AC5 group. The density of neurons stained by Cresyl violet was increased in all group without control group. The density of ChAT was increased in AC2, ACS group. Conclusions : Our study suggests that AC5 group show anti-apoptotic and neuroprotective effects on cholinergic neuron in focal cerebral ischemia of the stroke in rats.

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Molecular cloning and characterization of β-1,3-glucanase gene from Zoysia japonica steud (들잔디로부터 β-1,3-glucanase 유전자의 클로닝 및 특성분석)

  • Kang, So-Mi;Kang, Hong-Gyu;Sun, Hyeon-Jin;Yang, Dae-Hwa;Kwon, Yong-Ik;Ko, Suk-Min;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.43 no.4
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    • pp.450-456
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    • 2016
  • Rhizoctonia leaf blight (large patch) has become a serious problem in Korean lawn grass, which is extremely hard to treat and develops mostly from the roots of lawn grass to wither it away. Rhizoctonia leaf blight (large patch) is caused by Rhizoctonia solani AG2-2 (IV). To develop zoysia japonica with strong disease tolerance against this pathogenic bacterium, ${\beta}-1,3-glucanase$ was cloned from zoysia japonica, which is one of the PR-Proteins known to play a critical role in plant defense reaction. ${\beta}-1,3-glucanase$ is known to be generated within the cells when plant tissues have a hypersensitive reaction due to virus or bacterium infection and secreted outside the cells to play mainly the function of resistance against pathogenic bacteria in the space between the cells. This study utilized the commonly preserved part in the sequence of corn, wheat, barley, and rice which had been researched for their disease tolerance among the ${\beta}-1,3-glucanase$ monocotyledonous plants. Based on the part, degenerate PCR was performed to find out the sequence and full-length cDNA was cloned. E.coli over-expression was conducted in this study to mass purify target protein and implement in vitro activation measurement and antibacterial test. In addition, to interpret the functions of ZjGlu1 gene, each gene-incorporating plant transformation vectors were produced to make lawn grass transformant. Based on ZjGlu1 protein, antibacterial activity test was conducted on 9 strains. As a result, R. cerealis, F. culmorum, R.solani AG-1 (1B), and T. atroviride were found to have antibacterial activity. The gene-specific expression amount in each organ showed no huge difference in the organs based upon the transformant and against 18s gene expression amount.

Site-directed Mutagenesis of Arginine 221 of Lymphocyte Mono-ADP-ribosyltransferase (임파구 Mono ADP-ribosyltransferase의 Arginine 221의 자리 지정 돌연변이의 유발)

  • Hyun-Ju Kim
    • Journal of Life Science
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    • v.8 no.5
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    • pp.486-491
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    • 1998
  • Previously, we have cloned and characterized two ADP-ribosyltransferases (Yac-1 and Yac-2) from mouse Iym-phocyte. Yac-2 transferase contains significant NAD glycohydrolase activity as well as ADP-ribosyltransferase acti-vity. Yac-2 has an arginine at position 221 between two conserved glutamic acids. To investigate the significance of Arg-221 on enzyme activities, Arg-221 was mutagenized to Glu (R221E) and to Ala (R221A). Mutants R221E and R221A were active as wild type for ADP-ribosyltransferase and NAD glycohydrolase activity, suggesting that the arginine 221 in Yac-2 does not play a major role in enzyme activities.

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Inhibitory and Excitatory Postsynaptic Currents of Medial Vestibular Nucleus Neurons of Rats

  • Chun, Sang-Woo;Choi, Jeong-Hee;Park, Byung-Rim
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.2
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    • pp.59-63
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    • 2003
  • The medial vestibular nucleus (MVN) neurons are controlled by excitatory synaptic transmission from the vestibular afferent and commissural projections, and by inhibitory transmission from interneurons. Spontaneous synaptic currents of MVN neurons were studied using whole cell patch clamp recording in slices prepared from 13- to 17-day-old rats. The spontaneous inhibitory postsynaptic currents (sIPSCs) were significantly reduced by the $GABA_A$ antagonist bicuculline ($20{\mu}M$), but were not affected by the glycine antagonist strychnine ($1{\mu}M$). The frequency, amplitude, and decay time constant of sIPSCs were $4.3{\pm}0.9$ Hz, $18.1{\pm}2.0$ pA, and $8.9{\pm}0.4$ ms, respectively. Spontaneous excitatory postsynaptic currents (sEPSCs) were mediated by non-NMDA and NMDA receptors. The specific AMPA receptor antagonist GYKI-52466 ($50{\mu}M$) completely blocked the non-NMDA mediated sEPSCs, indicating that they are mediated by an AMPA-preferring receptor. The AMPA mediated sEPSCs were characterized by low frequency ($1.5{\pm}0.4$ Hz), small amplitude ($13.9{\pm}1.9$ pA), and rapid decay kinetics ($2.8{\pm}0.2$ ms). The majority (15/21) displayed linear I-V relationships, suggesting the presence of GluR2-containing AMPA receptors. Only 35% of recorded MVN neurons showed NMDA mediated currents, which were characterized by small amplitude and low frequency. These results suggest that the MVN neurons receive excitatory inputs mediated by AMPA, but not kainate, and NMDA receptors, and inhibitory transmission mediated by $GABA_A$ receptors in neonatal rats.

Study on the Relationship between Skin Dryness and Amino Acids in Stratum Corneum (아미노산 동시분석을 통한 피부보습능과 각질 중 아미노산 함량과의 상관관계 연구)

  • Joo, Kyung-Mi;Han, Ji-Yeon;Son, Eui-Dong;Nam, Gae-Won;Jeong, Hye-Jin;Lim, Kyung-Min;Cho, Jun-Cheol
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.1
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    • pp.75-82
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    • 2012
  • Natural moisturizing factors (NMFs) are hydrophilic and water-soluble components in stratum corneum of the skin. NMFs absorb water from outer environment and enhance the water-holding capacity of stratum corneum and thereby, prevent the dryness and increase flexibility and plasticity of skin. NMFs are mainly composed of amino acids and their metabolites that are produced from the degradation of filaggrin. Here we established a simultaneous quantification method for 22 amino acids in tape-stripped stratum corneum samples using UPLC-PDA. With this method, we analyzed amino acid contents from tape-stripped stratum corneum samples of forearm and forehead regions from 15 healthy volunteers. Amino acid contents of inner (or upper) region were higher than outer (or lower) region of stratum corneum. Amino acid contents of stratum corneum of forearm were higher by 1.5 fold than forehead region. Of note, total amino acid contents were significantly and inversely correlated with trans-epidermal water loss (TEWL), an index for skin barrier function. Especially, Ser, Glu, Gly, Ala and Thr were determined to positively affect skin mositurizing activities. In conclusion, we could demonstrate that amino acid contents of stratum corneum are closely linked with skin barrier and moisturizing function, providing an important and fundamental methodology for the study of amino acids in skin physiology.