• Title/Summary/Keyword: $\Ca^{2+}$-ATPase

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Effect of Refrigerated Sea Water on Keeping Freshness of Sardine (냉각해수저장법에 의한 정어리의 선도유지)

  • CHO Young-Je;KIM Chung-Gon;LEE Kang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.21 no.3
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    • pp.177-183
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    • 1988
  • The Present study has been carried out to investigate the effect of refrigerated seawater (RSW) at $-1^{\circ}C$ in order to delay spoilage and extend the shelf-life in Sardine(Sardinops melanosticta). The result obtained are summarized as follows : In case of freshness, K-value was shown above $30\%$ at 1 day of storage by icing, but in RSW system was shown $20\%$ until 2 days by storage. VBN and TMA were also shown the similar tendency when compared to K-value. The lipids extracted from sardine muscle held in ice was oxidized much more than those in RSW system by way of chemical test such as TBA and POV. The myofibrillar protein extracted from sardine stored in RSW system was denaturized more slowly compared with those by ice. $Ca^{2+}-ATPase$ activity indicated that myofibrillar protein held in RSW system was more stable than those held in ice. Total viable counts for sardine in RSW system showed an overall lower values. By TPA(Textural Porfile Analysis) , sardine meat pastes held in RSW system have a higher value than those held in ice.

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Effect of Quercus acutissima CARRUTHERS Extracts on the Lipid Metabolism (도토리추출물이 흰쥐의 체내 지질대사에 미치는 영향)

  • Sung, In-Suk;Kim, Myung-Joo;Cho, Soo-Yeul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.2
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    • pp.327-333
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    • 1997
  • This study was conducted to investigate the effect of Quercus acutissima CARRUTHERS extracts on lipid metabolism. Sprague-Dawley male rats$(110{\pm}10g)$ were fed on containing normal and high fat diets. They were orally administrated(0.02g/100g B.W.) of Quercus aculissima CARRUTHERS ethylacetate-extract and water-extract at the same time once a day respectively. The rats were sacrificed after 6 weeks of feeding periods. In high fat diet group, liver and heart weight were increased but kidney weight was decreased. Contents of total lipid, triglyceride and phospholipid were increased in high fat diet groups. But the degree of increment was reduced by administration of Quercus acutissima CARRUTHERS extracts and water extract was more effective. Significant decrease in serum total lipid content by administration Quercus acutissima CARRUTHERS extracts was not due to decrease of triglyceride content but total cholesterol content. Whereas HDL-cholesterol content was significantly decreased in high fat diet group and improved by administration of Quercus acutissima CARRUTHERS extracts. Total lipid, triglyceride and total cholesterol contents in liver were also increased in high fat diet group but phospholipid content was significantly decreased. The results indicate that Quercus acutissima CARRUTHERS extracts were effective in preventing hyperlipidemia and water extract was more effective.

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Identification of Calcium/Calmodulin-Dependent Phosphatase as the Dephosphorylating Enzyme of IgE-Dependent Histamine-Releasing Factor in RBL-2H3 (RBL-2H3 세포에서 IgE-depnedent Histamine-releasing Factor의 탈인산화 효소에 관한 연구)

  • Hwang Sun-Ok;Lee Kyunglim
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.189-193
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    • 2005
  • IgE-dependent histamine-releasing factor(HRF) was initially described as a secretagogue for secretion of histamine from IgE+ basophils from a subset of allergic donors. Previously, we identified that S98 residue of HRF was phosphorylated using anti-HRFpS98 antibody which specifically recognizes the phosphorylated serine residue of HRF and HRFS98A mutant construct. In vitro kinase assay, only wild type HRF was phosphorylated by PKC, and S98A HRF was not affected by PKC. In this study, we attempted to characterize the phosphatase which specifically dephosphorylates HRF by immunoprecipitation and pull-down assay. In RBL-2H3 cells, HRF interacted only with calcineurin (also called as PP2B, calcium/calmodulin-dependent phosphatase) but not with PP1 or PP2A. The results suggest that HRF is most likely dephosphory-lated by calcineurin.

Isolation and Characterization of Paramyosins of Marine Gastropods (해산복족류(海産腹足類)의 Paramyosin의 분리(分離) 및 그 특성(特性)에 관(關)한 연구(硏究))

  • Pyeun, Jae-Hyeung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.2 no.1
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    • pp.1-21
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    • 1973
  • The muscle of abalone, Notohaliotis discus (REEVE), and top-shell, Turbo cornutus Solander, were examined for protein composition. Then paramyosins which are known as one of the important structural protein of the muscle fibrils were isolated from the both muscle and their physico-chemical properties such as solubility, salting-out behaviour, intrinsic viscosity, ATPase activity, etc. involving amino acid composition and N-terminal amino acid residues were investigated to elucidate phylogenie characteristics more intensively from the viewpoint of comparative biochemistry. The analysis of protein composition resulted in the following estimations: abalone muscle; water-soluble protein of 22 %, salt-soluble protein, 34%, alkali-soluble protein, 20%, and stroma protein, 24%, and top-shell muscle; water-soluble protein of 16%, salt-soluble protein, 30%, alkali-soluble protein, 29%, and stroma protein, 25%, respectively. It is demonstrated in sedimentation analysis that paramyosin and myosin-actomyosin account for approximately 65% and 35% of the salt-soluble protein of abalone, and that the composition of both sediments in top-shell was approximately 70% and 30%, respectively. The ultracentrifugally homogenous paramyosins isolated essentially according to Bailey's ethanol-dried method from both of the muscle showed a $S^{\circ}_{20,w}$ of 3. 14s for abalone and a $S^{\circ}_{20,w}$ of 3.50s for top-shell. The both paramyosins were commonly rich in arginine, aspartic acid, and glutamic acid, while scarcely contained proline and tryptophan, in rough accord with the other paramyosins thus far reported. It is clear that these gastropod paramyosins showed of having the characteristic N-terminal amino acid residues such as N-aspartic acid, N-valine, N-serine, and N-threonine in common. The abalone paramyosin completely salted in with KCl beyond $0.35{\mu}$ and the top-shell paramyosin beyond $0.30{\mu}$. The abalone paramyosin was salted-out between 18% and 30% saturation of ammonium sulphate and the top-shell paramyosin between 22% and 29% saturation. The intrinsic viscosities at abalone and top-shell paramyosins at $25^{\circ}C$ were estimated respectively to be 3.1 dl/g and 2.6 dl/g showing somewhat higher than the values for some other paramyosins from lamellibranchs. In regard with the ATPase activity, the para myosin specimens did not exhibit any significant activity over through the pH conditions of 5 to 9.5. irrespective of the presence of $Ca^{++}$ or $Mg^{++}$. So was the case with the abalone paramyosin prepared by a slightly modified Bailey's wet-extraction method.

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[ $P2X_2$ ] Receptor Activation Potentiates PC12 Cell Differentiation Induced by ACAP in Acidic Environments

  • Lee, Myung-Hoon;Nam, Jin-Sik;Ryu, Hye-Myung;Yoo, Min;Lee, Moon-Hee
    • Biomedical Science Letters
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    • v.13 no.3
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    • pp.197-206
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    • 2007
  • P2X receptors are membrane-bound ion channels that conduct $Na^+,\;K^+$, and $Ca^{2+}$ in response to ATP and its analogs. There are seven subunits identified so far ($P2X_1-P2X_7$). $P2X_2$ receptors are known to be expressed in a wide range of organs including brains and adrenal grands. PC12 cells are originated from adrenal grand and differentiated by nerve growth factor or pituitary adenylate cyclase activating poly peptide (PACAP). Previous studies indicate that $P2X_2$ receptor activation in PC12 cells couples to $Ca^{2+}-dependent$ release of catecholamine and ATP. It is known that acidic pH potentiates ATP currents at $P2X_2$ receptors. This leads to a hypothesis that $P2X_2$ receptors may play an important role in PC12 cell differentiation, one of the characteristics of which is neurite outgrowth, induced by the hormones under lower pH. In the present study, we isolated several clones which potentiate neurite outgrowth by PACAP in acidic pH (6.8), but not in alkaline pH (7.6). RT-PCR and electrophysiology data indicate that these clones express only functional $P2X_2$ receptors in the absence or presence of PACAP for 3 days. Potentiation of neurite outgrowth resulted from PACAP (100 nM) in acidic pH is inhibited by the two P2X receptor antagonists, suramin and PPADS ($100\;{\mu}M)$ each), and exogenous exprerssion of ATP-binding mutant $P2X_2$ receptor subunit ($P2X_2[K69A]$). However, acid sensing ion channels (ASICs) are not involved in PACAP-induced neurite outgrowth potentiation in lower pH since treatments of an inhibitor of ASICs, amyloride ($10\;{\mu}M$), did not give any effects to neurite extension. The vesicular proton pump ($H^+-ATPase$) inhibitor, bafilomycin (100 nM), reduced neurite extension indicating that ATP release resulted from $P2X_2$ receptor activation in PC12 cells is needed for neurite outgrowth. These were confirmed by activation of mitogen activated protein kinases, such as ERKs and p38. These results suggest roles of ATP and $P2X_2$ receptors in hormone-induced cell differentiation or neuronal synaptogenesis in local acidic environments.

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Comparative Effects of Prostaglandin $F_2$ alpha and Ouabain on the Isolated Rat Atria (Rat적출심방 운동성에 대한 Prostaglandin $F_2$ alpha와 Ouabain작용의 비교)

  • Lee Kwang-Youn
    • The Korean Journal of Pharmacology
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    • v.20 no.1 s.34
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    • pp.55-65
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    • 1984
  • Comparative effects of $PGF_{2{\alpha}}$ and ouabain on the isolated rat(Sprague-Dowley) atria were studied. The isolated rat atria were prepared for isometric myography in the isolated organ bath containing Feigen's solution perfused with 95% $O_2$ and 5% $CO_2$, and the pH of the medium was maintained at 7.4. The cumulative concentration-response relationship revealed the positive inotropic effects of both drugs with the higher potency of $PGF_{2{\alpha}}$ and the higher efficacy of ouabain. $PGF_{2{\alpha}}$ showed a positive chronotropic effect, but ouabain showed a tendency of increasing the contraction rate. In low-Ca(1.4 mM) medium, the positive inotropic and chronotropic effect of $PGF_{2{\alpha}}$(by $3{\times}10^{-8}M$) were preponderant $(p<0.05{\sim}p<0.005)$ over those of ouabain(by $3{\times}10^{-3}M$). $Ca^{++}$-addition(cumulative, to 2.8, 4.2, 5.6, and 7.0 mM) into the medium evoked the more sensitive response in the $PGF_{2{\alpha}}$ group than in the ouabain group. In low-K(2.8 mM) medium, the $PGF_{2{\alpha}}a(3{\times}10^{-8}M)$ group and the ouabain$(3{\times}10^{-3}M)$ group showed similar tensions(DT and RT) and contraction rates. And both group showed significantly(p<0.05p<0.01) higher tensions and contraction rates than those of the control group. By the cumulative addition of the $K^+$(to 4.2, 5.6, 7.0 and 8.4 mM), only the DT of the $PGF_{2{\alpha}}$ group was sustained at signifcantly$(p<0.05{\sim}p<0.01)$ higher level than the DT of the control group. The $K^+$-addition inhibited the positive inotropic effect of ouabain significantly (p<0.05). The cumulative addition of lidocaine in high concentrations $(1{\times}10^{-5}\;to\;1{\times}10^{-3}M)$ evoked no significant influence on the intropic activities of $PGF_{2{\alpha}}$ and ouabain, but significant ${\beta}$-blockade with propranolol could not inhibit the positive intropic and chronotropic effect of $PGF_{2{\alpha}}$. In conclusion, it is presumed that $PGF_{2{\alpha}}$ may have some more active mechanism of accelerating the influx of $Ca^{++}$ across the cell membrane of the isolated rat atria as compared with ouabain, and the action site may be located at the cell membrane. As a supposition which needs further investigations, it is presumed that $PGF_{2{\alpha}}$ may have its specific membrane receptors on the atrial muscle or sinus node cells.

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Peptidoglycan Induces the Production of Interleukin-8 via Calcium Signaling in Human Gingival Epithelium

  • Son, Aran;Shin, Dong Min;Hong, Jeong Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.1
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    • pp.51-57
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    • 2015
  • The etiology of periodontal disease is multifactorial. Exogenous stimuli such as bacterial pathogens can interact with toll-like receptors to activate intracellular calcium signaling in gingival epithelium and other tissues. The triggering of calcium signaling induces the secretion of pro-inflammatory cytokines such as interleukin-8 as part of the inflammatory response; however, the exact mechanism of calcium signaling induced by bacterial toxins when gingival epithelial cells are exposed to pathogens is unclear. Here, we investigate calcium signaling induced by bacteria and expression of inflammatory cytokines in human gingival epithelial cells. We found that peptidoglycan, a constituent of grampositive bacteria and an agonist of toll-like receptor 2, increases intracellular calcium in a concentration-dependent manner. Peptidoglycan-induced calcium signaling was abolished by treatment with blockers of phospholipase C (U73122), inositol 1,4,5-trisphosphate receptors, indicating the release of calcium from intracellular calcium stores. Peptidoglycan-mediated interleukin-8 expression was blocked by U73122 and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethyl ester). Moreover, interleukin-8 expression was induced by thapsigargin, a selective inhibitor of the sarco/endoplasmic reticulum calcium ATPase, when thapsigargin was treated alone or co-treated with peptidoglycan. These results suggest that the gram-positive bacterial toxin peptidoglycan induces calcium signaling via the phospholipase C/inositol 1,4,5-trisphosphate pathway, and that increased interleukin-8 expression is mediated by intracellular calcium levels in human gingival epithelial cells.

Study on Meat Tenderizer -Part II. Tenderizing ability of Enzyme from Asp. oryzae- (Meat Tenderizer 제조에 관한 연구 -제2보 Asp. oryzae 생산 protease의 연육효과-)

  • Lee, Jung-Hee;Kim, Kun-Wha;Yu, Ju-Hyun;Yang, Ryung
    • Korean Journal of Food Science and Technology
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    • v.7 no.4
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    • pp.229-237
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    • 1975
  • An attempt was made to utilize the enzyme produced by Asp. oryzae as meat tenderizer. The production, purification, and various properties of proteinase produced by Asp. oryzae were investigated. Results obtained are as follow; 1. A strain which had the highest proteolytic activity was selected among 9 Aspergillus species. 2. Culture medium consisted of wheat bran 10g, 2% glucose, 0.03% urea and 0.1% $MgSO_4$ (pH 6.5). Mold was incubated at $30^{\circ}C$ for 3 days. 3. Enzyme extract from culture medium were fractionated with ammonium sulfate and purified by Sephadex G-75 column chromatography. 4. When pH of reaction mixture was controlled, maximal activity of proteinase by Asp. oryzae was obtained at pH 3, pH 6.6, $8.4{\sim}8.5$ and pH 10.0 to 10.5. Those results were interpreted to show that enzyme consists of acid proteinase, neutral proteinase and alkaline proteinase. Enzyme was stable at pH 6 to 10. 5. Opt. temperature for proteinase activity was $50^{\circ}C$, but enzyme was stable up to $40^{\circ}C$. 6. The proteinase was inhibited by $Ag^+$. It was also inhibited by EDTA. 7. When myofibrillar proteins were treated by proteinase from Asp. oryzae, ATPase activities of myofibrillar proteins changed remarkably. Accordingly, it was concluded that proteinase produced by Asp. oryzae were able to be used as meat tenderizer.

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${\alpha}$-Mangostin Reduced ER Stress-mediated Tumor Growth through Autophagy Activation

  • Kim, Sung-Jin;Hong, Eun-Hye;Lee, Bo-Ra;Park, Moon-Ho;Kim, Ji-Won;Pyun, A-Rim;Kim, Yeon-Jeong;Chang, Sun-Young;Chin, Young-Won;Ko, Hyun-Jeong
    • IMMUNE NETWORK
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    • v.12 no.6
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    • pp.253-260
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    • 2012
  • ${\alpha}$-Mangostin is a xanthon derivative contained in the fruit hull of mangosteen (Garcinia mangostana L.), and the administration of ${\alpha}$-Mangostin inhibited the growth of transplanted colon cancer, Her/CT26 cells which expressed Her-2/neu as tumor antigen. Although ${\alpha}$-Mangostin was reported to have inhibitory activity against sarco/endoplasmic reticulum $Ca^{2+}$ ATPase like thapsigargin, it showed different activity for autophagy regulation. In the current study, we found that ${\alpha}$-Mangostin induced autophagy activation in mouse intestinal epithelial cells, as GFP-LC3 transgenic mice were orally administered with 20 mg/kg of ${\alpha}$-Mangostin daily for three days. However, the activation of autophagy by ${\alpha}$-Mangostin did not significantly increase OVA-specific T cell proliferation. As we assessed ER stress by using XBP-1 reporter system and phosphorylation of $eIF2{\alpha}$, thapsigargin-induced ER stress was significantly reduced by ${\alpha}$-Mangostin. However, coadministration of thapsigargin with ${\alpha}$-Mangostin completely blocked the antitumor activity of ${\alpha}$-Mangostin, suggesting ER stress with autophagy blockade accelerated tumor growth in mouse colon cancer model. Thus the antitumor activity of ${\alpha}$-Mangostin can be ascribable to the autophagy activation rather than ER stress induction.

The Effects of Gamigunshimtang on the Ischemic Heart Disease & Heart cell in Rats (허혈성심장(虛血性心臟) 및 심장세포(心臟細胞)에 대(對)한 가미건심탕(加味健心湯)의 실험적(實驗的) 연구(硏究))

  • Park, Jung-Mi;Moon, Sang-Kwan;Go, Chang-Nam;Cho, Gi-Ho;Kim, Kyung-Suk;Bae, Hyung-Sup;Lee, Kyung-Sup
    • The Journal of Korean Medicine
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    • v.19 no.1
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    • pp.251-270
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    • 1998
  • The effects of Gamigunshimtang on the isolated perfused ischemic heart in rats, heart rates, left ventricular pressure, cardiac blood flow and cardiotoxicity were stu.died in H9C2 myoblast cell, myocardial slice culture The results were as follows: 1. The administration of Gamigunshimtang to the rat recovered effectively heart rate, left ventricular pressure and flow rate from the experimental ischemia in perfused rat heart. The release of lactic dehydrogenase after the ischemia also decreased compared to the control group. 2. The administration of Gamigunshimtang to H9C2 myoblast culture enhanced the cell proliferation and protected against doxorubicin and allylamine induced release of the lactic dehydrogenase into the culture medium. It also protected effectively against doxorubicin and allylamine induced decrease of Ca ATPase activity and the increase of NADPH-cytochrome C reductase activity in the microsome. 3. The administration of Gamigunshimtang to the rat myocardial slice culture protected effectively against doxorubicin and allylamine induced decreases of protein synthesis and ATP content, and increases of cvtosolic enzyme, creatin kinase into the medium and lipid peroxidation.

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