• Title/Summary/Keyword: PLC

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Physiological Roles of Phospholipase Cγ and Its Mutations in Human Disease (Phospholipase Cγ의 생리적 기능과 질병과 연관된 돌연변이)

  • Jang, Hyun-Jun;Choi, Jang Hyun;Chang, Jong-Soo
    • Journal of Life Science
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    • v.30 no.9
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    • pp.826-833
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    • 2020
  • Phospholipase C gamma (PLCγ) has critical roles in receptor tyrosine kinase- and non-receptor tyrosine kinase-mediated cellular signaling relating to the hydrolysis of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] to produce inositol 1,4,5 trisphosphate (IP3) and diacylglycerol (DAG), which promote protein kinase C (PKC) and Ca2+ signaling to their downstream cellular targets. PLCγ has two isozymes called PLCγ1 and PLCγ2, which control cell growth and differentiation. In addition to catalytically active X- and Y-domains, both isotypes contain two Src homology 2 (SH2) domains and an SH3 domain for protein-protein interaction when the cells are activated by ligand stimulation. PLCγ also contains two pleckstrin homology (PH) domains for membrane-associated phosphoinositide binding and protein-protein interactions. While PLCγ1 is widely expressed and appears to regulate intracellular signaling in many tissues, PLCγ2 expression is restricted to cells of hematopoietic systems and seems to play a role in the regulation of immune response. A distinct mechanism for PLCγ activation is linked to an increase in phosphorylation of specific tyrosine residue, Y783. Recent studies have demonstrated that PLCγ mutations are closely related to cancer, immune disease, and brain disorders. Our review focused on the physiological roles of PLCγ by means of its structure and enzyme activity and the pathological functions of PLCγ via mutational analysis obtained from various human diseases and PLCγ knockout mice.

The Effects of Single Component of Ginsenosides on the Mechanism of Mediator Release in the Allergic Hypersensitivity (인삼 사포닌 단일물질이 알러지 과민반응의 매개체 유리기전에 미치는 영향)

  • Ro, Jai-Youl;Kim, Kyung-Hwan
    • The Korean Journal of Pharmacology
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    • v.30 no.2
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    • pp.243-254
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    • 1994
  • Inflammatory diseases, allergic and asthmatic disorders are caused by the mediator release from the activation of the phospholipase C (PLC), phospholipase D (PLD), methyltransferase or adenylate cyclase etc. during IgG or IgE cross-linking of high affinity receptors on mast cells or basophil surface. One important enzyme activated after IgG or IgE receptor cross-linking is PLD, the enzyme which converts phosphatidylcholine (PC) to phosphatidic acid (PA). Under the hypothesis that these may be some differences in mediator release according to the difference in PLD activity, we attempted to confirm the ginseng saponin effects on the PLD activity. We examined the PLD activity during the passively sensitized mast cell activation in the presence of single component of ginsenosides $(Rc,\;Rg_1,\;Rg_2,\;Rg_3)$. We also measured the amount of mediators (histamine and leukotrienes) released by stimulating with ovalbumin (OA) or calcium ionophore (CaI), Guinea Pig lung mast cells were purified using enzyme digestion, count current elutriation, and discontinuous Percoll density gradient. In purified mast cells prelabeled with $[^3H]$ arachidonic acid or $[^3H]$ palmitic acid, PLD activity was assessed more directly by the production of labeled PEt by PLD-mediated transphosphatidylation in the presence of ethanol. Histanine release was determined by Spectrophotofluorometry, and leukotrienes by radioimmunoassay. The PLD activity during the passively sensitized mast cell activation is increased up to $3{\sim}5times$. The PLD activity during the passively sensitized mast cell activation in the presence of all ginsenosides is decreased up to $4{\sim}11$ times. $Rg_l\;and\;Rg_2$ ginsenoside pretreatment decreased histamine and leukotrienes by 50% in the OA-induced or by 40% in the Cal-induced mast cell after passively sensitization. Rc pretreatment poorly decreased histamine but leukotrienes decreased by 70% in the OA-induced or by 35% in the Cal-induced mast cell. $Rg_3$ ginsenoside pretreatment increased histamine release without challenging OA or Cal but leukotrienes decreased. These observations indicate that single unit of ginsenosldes may be an important contributor to inhibit the release of histamine and leukotrienes in the guinea pig lung mast cells, that inhibits the PLD-mediated formation of DAG evoked by mast cell activation.

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Rehmannia Glutinosa Pharmacopuncture Solution Regulates Functional Activation, FcεRI Expression, and Signaling Events in Mast Cells

  • Kang, Kyung-Hwa;Lee, Kyung-Hee;Yoon, Hyun-Min;Jang, Kyung-Jeon;Song, Chun-Ho;Kim, Cheol-Hong
    • Journal of Pharmacopuncture
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    • v.15 no.4
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    • pp.32-41
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    • 2012
  • Objectives: Rehmannia glutinosa pharmacopuncture solution (RGPS) was investigated to determine both its anti-allergic inflammatory effects on mast cells and its detailed mechanism of actions. Methods: We investigated whether RGPS suppress cytokines, enzymes, $Fc{\varepsilon}RI$ expression and $Fc{\varepsilon}RI$-mediated signaling in RBL-2H3 cells stimulated with anti-DNP IgE/DNP-HSA. The suppressive effects of RGPS on the levels of cytokines such as IL-$1{\beta}$, IL-6 and GM-CSF were measured using emzyme-linked immunospecific assay (ELISA). The mRNA expression levels of cytokines, enzymes (HDC2, COX-1, COX-2 and 5LO) and $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunits were measured using reverse transcription polymerase chain reaction (RT-PCR) method. The activation of $Fc{\varepsilon}RI$-mediated signaling was examined using Western blot analyses. Results: RGPS suppressed production of proinflammatory cytokines (IL-$1{\beta}$, IL-6, and GM-CSF) in stimulated RBL-2H3 cells significantly (p < 0.05). RGPS also suppressed mRNA expression of inflammatory enzymes (HDC2, COX-1, COX-2, 5LO). In addition, mRNA expression levels of $Fc{\varepsilon}RI{\alpha}$, $Fc{\varepsilon}RI{\beta}$and $Fc{\varepsilon}RI{\gamma}$ were lowered by treatment with RGPS. Finally, RGPS prevented phosphrylation of Lyn, Syk, LAT, Gab2, PLC ${\gamma}1/2$, PI3K, Akt, cPLA2 and $I{\kappa}B{\alpha}$. Conclusions: RGPS effectively suppresses mast cell activations such as degranulation and inflammatory response via down-regulation of the $Fc{\varepsilon}RI$-mediated signaling pathways in IgE/Ag-stimulated mast cells.

Development of a Prototype Automatic Sorting System for Dried Oak Mushrooms (건표고 자동선별을 위한 시작시스템 개발)

  • Hwang, H.;Lee, C.H.
    • Journal of Biosystems Engineering
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    • v.21 no.4
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    • pp.414-421
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    • 1996
  • 한국과 일본의 경우 건표고를 외관의 품질상태 에 따라 12등급에서 16등급으로 구분하고 있다. 그리고 등급판정 작업은 임의로 추출한 샘플을 대상으로 전문 감정가에 의해 수작업으로 수행되고 있다. 건표고의 품질을 결정짓는 외관의 품질인자들은 갓과 내피에 고루 분포하고 있다. 본 논문에서는 컴퓨터 영상처리 시스템에 의거하여 개발한 건표고 자동 등급판정 및 선별 시작시스템의 구조와 기능 그리고 성능에 대하여 설명하였다. 개발한 시작시스템은 표고의 이송과 취급자동화를 위한 진동이송기, 반전장치, 컨베이어 이송장치와 두 세트의 컴퓨터 영상처리 시스템, 그리고 시스템 통괄제어를 위한 IBM PC AT호환 컴퓨터, 디지털 입출력 보드, 전공압실린더 구동제어를 위한 PLC등으로 구성하였다. 등급판정의 효율성 및 실시간 작업시스템을 고려하여 건표고의 등급판정은 두 세트의 컴퓨터 영상처리 시스템을 이용하여 이송되는 건표고의 갓 또는 내피 중 어디가 위를 향하는 지에 따라 두 단계에 걸쳐 독립적으로 판정을 수행하도록 하였다. 첫 번째 영상처리부에서는 갓표면 영상으로부터 4등급의 고품질 표고를 분류하며 두 번째 영상처리부에서는 내피표면 영상으로부터 중간 및 저품질 표고를 8개의 등급으로 분류한다. 실시간 영상정보처리를 목적으로 기존에 개발한 신경회로망을 이용한 등급판정 알고리즘을 시작시스템에 적용하였다. 개발한 시작기는 88% 이상의 등급판정 정확도를 보여 주었으며, 전공압시스템의 구동제약으로 인하여 표고 1개당 약0.7초의 선별시간이 소요되었다. 일조 선별라인의 경우 본 연구에서 제안한 시작기의 선별능력은 표고가 일차 처리부로 갓이 위로 올라와 있는 상태로 계속 공급된다면 시간당 대략 5,000여 개의 표고를 처리할 수 있을 것으로 기대된다.보강하여 가능하면 B-Pillar의 Middle이 Bending type collapse을 방지하여 Pelvis와 Door가 먼저 접촉하는 방법 등이 적용가능하다. 제작하기 이전에 설계된 부품에 대한 스프링 상수 및 내구특성을 체계적으로 규명하여 제품 시험의 횟수를 줄이고, 보다 정밀한 제품을 제작할 수 있도록 하기 위한 것이다.세포수는 초기 배반포기배에서 팽윤 배반포기배로 진행됨에 따라 두배에서 세배 정도 증가되었음을 알 수 있었다. 또한, differential labelling과 bisbenzimide기법에서 얻어진 각각의 총세포수를 비교하였을 때 총세포수는 발달의 진행 정도에 따라 증가되며 그와 동시에 동일한 군 간의 세포수도 거의 유사함을 알 수 있었다. 따라서, ICM과 TE를 differential labelling하는 기법은 수정란의 quality를 평가하는데 매우 유용한 기법으로서 착상전 embryo 발달을 연구하는데 효과적으로 이용될 수 있다는 것을 시사한다. 고도의 유의차를 나타낸 반면 비수구, 초생수로구 및 Bromegrass 목초구 간에는 아무런 유의차가 인정되지 않았다. 7. 농지보전 처리구인 배수구와 초생수로구는 비처리구에 비해 낮은 침두 유출량과 낮은 토양유실량을 나타내었다.구보다 14% 절감되는 것으로 나타났다.작용하는 것으로 사료된다.된다.정량 분석한 결과이다. 시편의 조성은 33.6 at% U, 66.4 at% O의 결과를 얻었다. 산화물 핵연료의 표면 관찰 및 정량 분석 시험시 시편 표면을 전도성 물질로 증착시키지 않고, Silver Paint 에 시편을 접착하는 방법으로도 만족한 시험 결과를 얻을 수 있었다.째, 회복기 중에 일어나는 입자들의 유입은 자기폭풍의 지속시간을 연장시키는 경향을 보이며 큰

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The Inhibitory Mechanism of Aloe Glycoprotein (NY945) on the Mediator Release in the Guinea Pig Lung Mast Cell Activated with Antigen-Antibody Complexes

  • Ro, Jai-Youl;Lee, Byung-Chul;Chung, Myung-Hee;Lee, Seung-Ki;Sung, Chung-Ki;Kim, Kyung-Hwan;Park, Young-In
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.1
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    • pp.119-131
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    • 1998
  • It has been reported that the glycoprotein extracted from Aloe has strong anti-inflammatory response. However, there has been no research report yet about the effect of Aloe on allergic hypersensitivity reactivity. By using guinea pig lung mast cells, this study aimed to examine the effects of Aloe glycoprotein (NY945) on the mediator releases caused by mast cell activation, and also aimed to assess the effects of NY945 on the mechanism of mediator releases in the mast cell activation. We partially purified mast cell from guinea pig lung tissues by using the enzyme digestion, the rough and the discontinuous density percoll gradient method. Mast cells were sensitized with IgG1 (anti-OA) and challenged with ovalbumin. Histamine was assayed by fluorometric analyzer, leukotrienes by radioimmunoassay. The phospholipase D activity was assessed by the production of labeled phosphatidylalcohol. The amount of mass 1, 2-diacylglycerol (DAG) was measured by the $[^3H]DAG$ produced when prelabeled with $[^3H]myristic$ acid. The phospholipid methylation was assessed by measuring the incorporation of the $[^3H]methyl$ moiety into phospholipids of cellular membranes. Pretreatment of NY945 (10 ${\mu}g$) significantly decreased histamine and leukotrienes releases during mast cell activation. The decrease of histamine release was stronger than that of leukotriene during mast cell activation. The phospholipase D activity increased by the mast cell activation was decreased by the dose-dependent manner in the pretreatment of NY945. The amount of DAG produced by PLC activity was decreased by NY945 pretreatment. The amount of mass 1, 2-diacylglycerol produced by activation of mast cells was decreased in the pretreatment of NY945. NY945 pretreatment strongly inhibited the incorporation of the $[^3H]methyl$ moiety into phospholipids. The data suggest that NY945 purified from Aloe inhibits in part an increase of 1, 2-diacylglycerol which is produced by activating mast cells with antigen-antibody reactions, which is mediated via phosphatidylcholine-phospholipase D and phosphatidylinositol-phospholipase C systems, and then followed by the inhibition of histamine release. Furthermore, NY945 reduces the production of phosphatidylcholine by inhibiting the methyltransferase I and II, which decreases the conversion of phosphatidylcholine into arachidonic acid and inhibits the production of leukotrienes.

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Listeria monocytogenes Serovar 4a is a Possible Evolutionary Intermediate Between L. monocytogenes Serovars 1/2a and 4b and L. innocua

  • Chen, Jianshun;Jiang, Lingli;Chen, Xueyan;Luo, Xiaokai;Chen, Yang;Yu, Ying;Tian, Guoming;Liu, Dongyou;Fang, Weihuan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.238-249
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    • 2009
  • The genus Listeria consists of six closely related species and forms three phylogenetic groups: L. monocytogenes-L. innocua, L. ivanovii-L. seeligeri-L. welshimeri, and L. grayi. In this report, we attempted to examine the evolutionary relationship in the L. monocytogenes-L. innocua group by probing the nucleotide sequences of 23S rRNA and 16S rRNA, and the gene clusters lmo0029-lmo0042, ascB-dapE, rplS-infC, and prs-ldh in L. monocytogenes serovars 1/2a, 4a, and 4b, and L. innocua. Additionally, we assessed the status of L. monocytogenes-specific inlA and inlB genes and 10 L. innocua-specific genes in these species/serovars, together with phenotypic characterization by using in vivo and in vitro procedures. The results indicate that L. monocytogenes serovar 4a strains are genetically similar to L. innocua in the lmo0035-lmo0042, ascB-dapE, and rplS-infC regions and also possess L. innocua-specific genes lin0372 and lin1073. Furthermore, both L. monocytogenes serovar 4a and L. innocua exhibit impaired intercellular spread ability and negligible pathogenicity in mouse model. On the other hand, despite resembling L. monocytogenes serovars 1/2a and 4b in having a nearly identical virulence gene cluster, and inlA and inlB genes, these serovar 4a strains differ from serovars 1/2a and 4b by harboring notably altered actA and plcB genes, displaying strong phospholipase activity and subdued in vivo and in vitro virulence. Thus, by possessing many genes common to L. monocytogenes serovars 1/2a and 4b, and sharing many similar gene deletions with L. innocua, L. monocytogenes serovar 4a represents a possible evolutionary intermediate between L. monocytogenes serovars 1/2a and 4b and L. innocua.

Development of Semi-automatic Cabbage Piling System for Tractor Implemented Chinese Cabbage Hervester (트랙터 부착형 배추 수확기용 반자동식 배추 적재시스템 개발)

  • Song, K. S.;Choi, D. Y.;Hwang, H.
    • Journal of Biosystems Engineering
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    • v.27 no.3
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    • pp.211-218
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    • 2002
  • 배추 생산에 있어서 수확, 운반, 적재 작업은 가장 노동이 집약적으로 요구되는 작업들이다. 최근, 여러 종류의 양배추 수화기가 일본과 유럽에서 개발되었다. 하지만 국내에서 재래되는 배추는 크기와 형태에 있어 양배추와는 달라 기 개발 기종의 도입이 어렵다. 또한 수확작업의 생력화 효과는 운반, 정선, 적재 작업과 밀접하게 연계되어 있어 출하시의 작업체계를 고려하여 수화에 따른 수집.반출 시스템을 개발하여야 한다. 수확시의 배추는 중량이 25~45 N 정도로 다 작물에 비하여 무겁고 부피가 크기 때문에 수확작업의 기계화를 위해서는 수확장치와 더불어 연속적으로 수확되는 배추를 적절하게 수집하여 적재하고 반출하는 시스템의 개발이 중요하다. 본 연구에서는 수확작업의 생력화 효과를 높이고 생력화 시스템 비용의 절감을 목적으로 작업자 1인에 의해 작업할 수 있는 반자동 형태의 트랙터 부착형 배추 수집, 적재, 반출시스템 시작기를 개발하였다. 시작기는 배추 이송장치, 적재장치, 팔렛 및 반출장치의 3개 부분과 PLC를 이용한 주 제어기로 구성하였다. 배추 수집용기로는 대략 70개의 배추를 담을 수 있는 크기가 1,050 mm$\times$1.050mm$\times$1,000mm 인 접이식 메쉬 팔렛을 사용하였으며 하단부에 롤러 안내판을 부착하여 적재한 팔렛의 배출이 용이하도록 하였다. 팔렛을 제외한 전체 시작기의 중량은 235 N 이였으며 크기는 3,940mm$\times$520mm$\times$1,630mm 이었다. 본 연구는 수확장치의 기능 및 생력화 효과를 극대화하고 배추의 손상정도를 최소화하는 시스템을 구성하고자 하였다. 이송장치는 트랙터 부착시 횡공간 점유율을 최소화하도록 하였으며 적재장치는 적재시 배추의 손상을 줄이고 배추가 놓이는 자세를 능동적으로 조절할 수 있도록 주름관을 부착하였다. 시작기의 실내시험 결과 이송장치는 0.18 m/s~0.36 m/s의 범위에서 적재장치는 0.4 m/s~2.4 m/s 범위에서 안정적으로 구동하였으며 두 장치를 동시에 구동하여 시험한 결과 이송장치는 0.26 m/s~0.36 m/s, 그리고 적재장치는 0.9 m/s~2.4 m/s 에서 적정하게 안정적으로 구동하였다. 적재장치의 성능에 있어서 1~3단 적재시에는 주름관을 이용하여 적재하고 4~5단 적재시에는 자유낙하에 의한 적재를 수행할 경우 인력에 의한 적재와 거의 동등한 적재량을 보였으며 손상정도는 거의 무시할 정도였다. 트랙터가 0.3 m/s로 주행하는 경우 노지로부터 배추를 뽑아 이송하는 뽑기벨트의 적정속도가 0.46 m/s인 점을 고려할 때 배추 이송 컨베이어는 0.34 m/s 이상의 속도를 유지할 필요가 있었으며 적재 컨베이어는 2 m/s~2.4 m/s의 속도에서 안정적으로 작동하였다. 배추의 주간 거리가 대략 30~40 cm 인 점을 감안하면 적재장치는 초당 1개의 적재성능을 보였다. 실내에서 수행한 시스템의 성능은 배추에 큰 손상없이 전반적으로 성공적으로 구동하였으나 향후 노면이 고르지 못한 포장에서의 성능 시험이 필요하다.

Development of a flower support for real flower decoration Automatic Production System (생화 장식 꽃받침 자동 생산 시스템 개발)

  • Song, Myung-Seok;Kim, Man-Joong;Kim, Seon-Bong;Ji, Peng;Ryuh, Beom-Sahng
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.7
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    • pp.63-71
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    • 2018
  • A flower support was developed for real flower decoration automation production system using an ultrasonic wave sealer to automatically produce a system. Because a flower support for real flower decoration that was produced manually could not meet the needs of the consumers, this study developed an automated manufacturing system to increase productivity. A flower support for real flower decoration was constructed using a cap consisting of plastic and plate made from non-woven fabric. The guide was designed to transport the cap to the ultrasonic wave sealer and optimal guide was developed from the test according to the material and shape. To produce the entire system, the guides and accessories were weighed and appropriate motors and pulleys were calculated. Control of the automation production system was based on a PCB board, which increased the reliability and security, and a remote controller with manual and automatic modes was prepared. After development, tests of the transfer precision and repetition accuracy revealed an X-axis of 2.7mm, a Y-axis of 1 mm, and a repetition of 0 mm. The productivity was also checked. The automated machine worked 8 hours/day to make 35 supports and 70 Therefore, the automatic system produces 200% more output than manual work

Acidic pH-activated $Cl^-$ Current and Intracellular $Ca^{2+}$ Response in Human Keratinocytes

  • Park, Su-Jung;Choi, Won-Woo;Kwon, Oh-Sang;Chung, Jin-Ho;Eun, Hee-Chul;Earm, Young-E;Kim, Sung-Joon
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.4
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    • pp.177-183
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    • 2008
  • The layers of keratinocytes form an acid mantle on the surface of the skin. Herein, we investigated the effects of acidic pH on the membrane current and $[Ca^{2+}]_c$ of human primary keratinocytes from foreskins and human keratinocyte cell line (HaCaT). Acidic extracellular pH ($pH_e{\leq}5.5$) activated outwardly rectifying $Cl^-$ current ($I_{Cl,pH}$) with slow kinetics of voltage-dependent activation. $I_{Cl,pH}$ was potently inhibited by an anion channel blocker 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS, 73.5% inhibition at 1${\mu}$M). $I_{Cl,pH}$ became more sensitive to $pH_e$ by raising temperature from $24^{circ}C$ to $37^{circ}C$. HaCaT cells also expressed $Ca^{2+}$-activated $Cl^-$ current ($I_{Cl,Ca}$), and the amplitude of $I_{Cl,Ca}$ was increased by relatively weak acidic $pH_e$ (7.0 and 6.8). Interestingly, the acidic $pH_e$ (5.0) also induced a sharp increase in the intracellular [$Ca^{2+}$] (${\triangle}[Ca^{2+}]_{acid}$) of HaCaT cells. The ${\triangle}[Ca^{2+}]_{acid}$ was independent of extracellular $Ca^{2+}$, and was abolished by the pretreatment with PLC inhibitor, U73122. In primary human keratinocytes, 5 out of 28 tested cells showed ${\triangle}[Ca^{2+}]_{acid}$. In summary, we found $I_{Cl,pH}$ and ${\triangle}[Ca^{2+}]_{acid}$ in human keratinocytes, and these ionic signals might have implication in pathophysiological responses and differentiation of epidermal keratinocytes.

Development of an Environmental Monitoring and Warning System for Cold Storage Rouse Using Internet (인터넷을 이용한 저온저장고 환경감시 및 경보 시스템 개발)

  • Jeong, Hoon;Yun, Hong-Sun;Lee, Won-Og;Cho, Kwang-Hwan;Cho, Young-Kil;Park, Won-Kyu;Shin, Jae-Hun
    • Food Science and Preservation
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    • v.10 no.1
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    • pp.28-31
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    • 2003
  • For safe storage of agricultural products in the cold storage house, accurate monitoring of temperature, humidity and gas conditions is necessary. This study was conducted to develop an environmental monitoring and warning system for the cold storage house to improve safety of storage. The system developed in this study is able to monitor temperature, humidity and $CO_2$concentration in the storage house and to send alarm signal to the farmer by telephone and beeper when abnormal conditions have been occurred in the storage house. And the developed system use internet network so we can supervise storage conditions in the home. From the results of the performance test, it was found that the temperature and relative humidity can be controlled within the range of 0.5$^{\circ}C$ and $\pm$2 percent. And farmer's response was fair.