• 대한약침학회지
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• 제16권1호
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• pp.43-49
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• 2013

Objective: Pyungwi-san (PWS) plays a role in a number of physiologic and pharmacologic functions in many organs. Interstitial cells of Cajal (ICCs) are pacemaker cells that generate slow waves in the gastrointestinal (GI) tract. We aimed to investigate the beneficial effects of PWS in mouse small-intestinal ICCs. Methods: Enzymatic digestion was used to dissociate ICCs from the small intestine of a mouse. The whole-cell patch-clamp configuration was used to record membrane potentials from the cultured ICCs. Results: ICCs generated pacemaker potentials in the GI tract. PWS produced membrane depolarization in the current clamp mode. Pretreatment with a $Ca^{2+}$-free solution and a thapsigargin, a $Ca^{2+}$-ATPase, inhibitor in the endoplasmic reticulum, eliminated the generation of pacemaker potentials. However, only when the thapsigargin was applied in a bath solution, the membrane depolarization was not produced by PWS. Furthermore, the membrane depolarizations due to PWS were inhibited not by U-73122, an active phospholipase C inhibitor, but by chelerythrine and calphostin C, protein kinase C inhibitors. Conclusions: These results suggest that PWS might affect GI motility by modulating the pacemaker activity in the ICCs.

• The Korean Journal of Physiology and Pharmacology
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• 제14권5호
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• pp.311-316
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• 2010

It is well-known that electrical field stimulation (EFS)-induced contraction is mediated by a cholinergic mechanism and other neurotransmitters. NO, ATP, calcitonin gene-related peptide (CGRP), and substance P are released by EFS. To investigate the purinergic mechanism involved in the EFS-induced contraction, purinegic receptors antagonists were used. Suramine, a non-selective P2 receptor antagonist, reduced the contraction induced by EFS. NF023 ($10^{-7}{\sim}10^{-4}M$), a selective P2X antagonist, inhibited the contraction evoked by EFS. Reactive blue ($10^{-6}{\sim}10^{-4}M$), selective P2Y antagonist, also blocked the contraction in a dose-dependent manner. In addition, P2X agonist ${\alpha}$,${\beta}$-methylene 5'-adenosine triphosphate (${\alpha}{\beta}MeATP$, $10^{-7}{\sim}10^{-5}M$) potentiated EFS-induced contraction in a dose-dependent manner. P2Y agonist adenosine 5'-[${\beta}$-thio]diphosphate trilithium salt ($ADP{\beta}S$, $10^{-7}{\sim}10^{-5}M$) also potentiated EFS-induced contractions in a dose-dependent manner. Ecto-ATPase activator apyrase (5 and 10 U/ml) reduced EFS-induced contractions. Inversely, 6-N,$N$-diethyl-D-${\beta}$,${\gamma}$- dibromomethylene 5'-triphosphate triammonium (ARL 67156, $10^{-4}M$) increased EFS-induced contraction. These data suggest that endogenous ATP plays a role in EFS-induced contractions which are mediated through both P2X-receptors and P2Y-receptors stimulation in cat esophageal smooth muscle.

• 한국식품과학회지
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• 제25권5호
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• pp.468-474
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• 1993

가공적성이 좋지 못한 적색육어류를 수산가공품의 효율적인 원료로 활용할 수 있는 새로운 가공법의 개발에 관한 연구로서 적색육어류의 하나인 고등어를 이용하여 고분자 수분흡수성 수지가 내재된 탈수시트로 저온에서 단시간 건조함으로써 건조공정 중 고도불포화지방산의 산화 및 단백질변성을 가능한 억제하여 안전성이 높은 반염건고등어의 제조를 시도하였고, 아울러 천일건조, 열풍건조 및 냉풍건조법으로 제조한 반염건고등어의 품질과 비교하였다. 반염건고등어의 제조를 위한 건조시간은 저온삼투압탈수법을 이용한 경우 $90{\sim}160$분으로 천일건조, 열풍건조 및 냉풍건조 등의 재래식 건조법을 이용하는 경우의 $180{\sim}510$분 보다 상당히 단축되었으며, 탈수시트의 종류에 따라서도 약간의 차이는 있어 sodium polyacrylate로 만든 탈수시트로 건조시킨 것이 가장 빨랐다. 반염건고등어의 수분함량은 $59.4{\sim}62.4%$의 범위이었고, 단백질함량 및 조지방함량은 각각 $26.3{\sim}28.1%$ 및 $7.2{\sim}9.7%$의 범위였다. 반염건고등어의 histamine함량은 $2.5{\sim}3.6mg/100g$의 범위로 건조방법에 관계없이 식중독은 문제되지 않았다. 반염건고등어의 과산화물값, 혼합지방산조성, 갈변도, 염용성질소의 용해도 및 근원섬유 Ca-ATPase 활성은 저온삼투압탈수법으로 제조한 것이 재래식 건조방법으로 제조한 것보다 변화가 적어 가공 중 지질산패 및 단백질변성이 억제되었다는 것을 알 수 있었다. 텍스튜어 및 관능검사 결과, 저온삼투압 탈수법으로 제조한 반염건고등어가 재래식 건조방법으로 제조한 반염건고등어보다 산패취가 적으면서 조직감이 우수하였다. 한편 탈수시트의 종류를 달리하여 저온삼투압탈수법으로 제조한 반염건고등어간에는 과산화물값, 혼합지방산조성, 염용성질소의 용해도 및 근원섬유 Ca-ATPase 활성에는 큰 차이가 없었다. 이상의 결과들로 미루어 볼 때 가공적성이 좋지 못하여 재래식 건조조건으로는 가공하기에 곤란하였던 적색육어류도 고분자 수분흡수제가 내재된 탈수시트로 탈수한다면 가공 중 지질산패, 단백질변성 및 바람직하지 못한 성분상호간의 반응을 최대한 억제할 수 있어 적색육어류의 식용화 뿐만 아니라 건조를 필요로 하는 수산식품가공에 광범위하게 이용 가능하리라는 결론을 얻었다.

• 한국양식학회지
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• 제6권2호
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• pp.107-123
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• 1993

조피볼락의 필수지방산인 n-3HUFA가 어체에 미치는 생화학적 변화를 알아보기 위하여, n-3HUFA 함량이 $0.0\~1.5\%$가 되도록 조정한 6종의 실험사료로 조피볼락 치어를 10주간 사육하여 혈액성상, 혈청성분 및 간세포 성상의 변화를 검토하였다. 혈액성상(RBC, Hb, Ht, MCV, MCHC 및 MCH)은 사료의 n-3HUFA 함량에 따라 유의적인 차이를 나타내지는 않았다. 혈청단백질 농도$(2.76\~3.63g/100ml)$ 및 혈당량$(36\~71mg/100ml)$은 n-3HUFA $0.9\%$ 사료까지는 증가하다가 그 이상에서는 일정한 값을 유지하였다. 혈중 total cholesterol 농도$(137\~86mg/100ml)$와 free cholesterol 농도$(84.2\~63.8mg/100ml)$는 n-3HUFA $1.2\%$까지 점차 감소하는 경향을 보였으며, 그 이상에서는 일정 하였다. 혈청 GPT $(159.7\~47.0 Karmen\; unit)$, GOT $(28.0\~11.1\;Karmen\;unit)$ 및 LDH $(3888\~1539\;W-unit)$도 모두 n-3HUFA $1.2\%$까지는 감소 경향을 보이다가 그 이후에는 일정한 값을 유지하였다. n-3HUFA 무첨가 사료를 먹은 실험어의 간조직은 n-3HUFA $1.5\%$ 첨가 사료를 먹은 실험어에 비해 인포색의 구조 붕괴와 부분적인 세포 융해가 관찰되었다. 간 cytosol의 LDH 활성과 microsomal membrane의 $Ca^{2+}-ATPase$ 활성도 변화되어 사료의 n-3HUFA가 증가할수록 증가한 경향을 보였다.

• 생명과학회지
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• 제24권8호
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• pp.915-926
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• 2014

리보솜은 mRNA상의 유전정보를 단백질로 번역하는 세포에 필수적인 거대복합체이다. 이러한 리보솜은 리보 핵산단백질 복합체로, rRNA와 리보솜 단백질로 이루어져있다. 리보솜 조립과정은 리보솜 단백질 이외에도 많은 조립인자들이 각 구성요소의 조립을 도움으로써 이루어진다. 세포 내 리보솜 조립과정에 참여하는 조립인자들로 GTPase, ATPase, 샤페론, RNA helicase, 수식효소 등 다양한 단백질들이 알려졌다. 리보솜 조립과정 중 이러한 조립인자들은 리보솜 단백질 또는 rRNA의 수식에 참여하거나, 리보솜 단백질들과 rRNA의 조립 등을 돕는다. 이러한 리보솜 조립인자들에 관한 유전학적, 구조적, 생화학적 실험결과들이 많이 존재하지만 정확한 리보솜 조립과정과 이러한 조립인자들의 역할에 대해서는 아직 밝혀지지 않았다. 현재까지의 연구결과를 바탕으로 E. coli의 리보솜 조립과정을 돕는 단백질들에 대하여 알아보고자 한다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5605-5611
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• 2012

Background: In the last two decades, pioneering research on anti-tumour activity of saffron has shed light on the role of crocetin, picrocrocin and safranal, as broad spectrum anti-neoplastic agents. However, the exact mechanisms have yet to be elucidated. Identification and characterization of the targets of bioactive constituents will play an imperative role in demystifying the complex anti-neoplastic machinery. Methods: In the quest of potential target identification, a dual virtual screening approach utilizing two inverse screening systems, one predicated on idTarget and the other on PharmMapper was here employed. A set of target proteins associated with multiple forms of cancer and ranked by Fit Score and Binding energy were obtained from the two independent inverse screening platforms. The validity of the results was checked by meticulously analyzing the post-docking binding pose of the picrocrocin with Hsp90 alpha in AutoDock. Results: The docking pose reveals that electrostatic and hydrogen bonds play the key role in inter-molecular interactions in ligand binding. Picrocrocin binds to the Hsp90 alpha with a definite orientation appropriate for nucleophilic attacks by several electrical residues inside the Hsp90-alpha ATPase catalytic site. Conclusion: This study reveals functional information about the anti-tumor mechanism of saffron bioactive constituents. Also, a tractable set of anti-neoplastic targets for saffron has been generated in this study which can be further authenticated by in vivo and in vitro experiments.

• The Korean Journal of Physiology and Pharmacology
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• 제17권2호
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• pp.149-156
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• 2013

Interstitial cells of Cajal (ICCs) are the pacemaker cells in the gastrointestinal tract, and histamine is known to regulate neuronal activity, control vascular tone, alter endothelial permeability, and modulate gastric acid secretion. However, the action mechanisms of histamine in mouse small intestinal ICCs have not been previously investigated, and thus, in the present study, we investigated the effects of histamine on mouse small intestinal ICCs, and sought to identify the receptors involved. Enzymatic digestions were used to dissociate ICCs from small intestines, and the whole-cell patch-clamp configuration was used to record potentials (in current clamp mode) from cultured ICCs. Histamine was found to depolarize resting membrane potentials concentration dependently, and whereas 2-PEA (a selective H1 receptor agonist) induced membrane depolarizations, Dimaprit (a selective H2-agonist), R-alpha-methylhistamine (R-alpha-MeHa; a selective H3-agonist), and 4-methylhistamine (4-MH; a selective H4-agonist) did not. Pretreatment with $Ca^{2+}$-free solution or thapsigargin (a $Ca^{2+}$-ATPase inhibitor in endoplasmic reticulum) abolished the generation of pacemaker potentials and suppressed histamine-induced membrane depolarization. Furthermore, treatments with U-73122 (a phospholipase C inhibitor) or 5-fluoro-2-indolyl des-chlorohalopemide (FIPI; a phospholipase D inhibitor) blocked histamine-induced membrane depolarizations in ICCs. On the other hand, KT5720 (a protein kinase A inhibitor) did not block histamine-induced membrane depolarization. These results suggest that histamine modulates pacemaker potentials through H1 receptor-mediated pathways via external $Ca^{2+}$ influx and $Ca^{2+}$ release from internal stores in a PLC and PLD dependent manner.

• Journal of Microbiology and Biotechnology
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• 제22권6호
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• pp.742-753
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• 2012

Copper-containing compounds are introduced into the environment through agricultural chemicals, mining, and metal industries and cause severe detrimental effects on ecosystems. Certain microorganisms exposed to these stressors exhibit molecular mechanisms to maintain intracellular copper homeostasis and avoid toxicity. We have previously reported that the soil bacterial isolate Achromobacter sp. AO22 is multi-heavy metal tolerant and exhibits a mer operon associated with a Tn21 type transposon. The present study reports that AO22 also hosts a unique cop locus encoding copper homeostasis determinants. The putative cop genes were amplified from the strain AO22 using degenerate primers based on reported cop and pco sequences, and a constructed 10,552 base pair contig (GenBank Accession No. GU929214). BLAST analyses of the sequence revealed a unique cop locus of 10 complete open reading frames, designated copSRABGOFCDK, with unusual separation of copCD from copAB. The promoter areas exhibit two putative cop boxes, and copRS appear to be transcribed divergently from other genes. The putative protein CopA may be a copper oxidase involved in export to the periplasm, CopB is likely extracytoplasmic, CopC may be periplasmic, CopD is cytoplasmic/inner membrane, CopF is a P-type ATPase, and CopG, CopO, and CopK are likely copper chaperones. CopA, B, C, and D exhibit several potential copper ligands and CopS and CopR exhibit features of two-component regulatory systems. Sequences flanking indicate the AO22 cop locus may be present within a genomic island. Achromobacter sp. strain AO22 is thus an ideal candidate for understanding copper homeostasis mechanisms and exploiting them for copper biosensor or biosorption systems.

• 대한한의학방제학회지
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• 제28권1호
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• pp.71-80
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• 2020

Obejectives : The purpose of this study was to investigate the effects of Yijin-tang on pacemaker potentials of small intestinal interstitial Cells of Cajal (ICC). Methods : To dissociate the ICC, we used enzymatic digestions from the small intestine in mice. The electrophysiological whole-cell patch-clamp configuration was used to record pacemaker potentials in the cultured ICC and the in vivo effects of Yijin-tang on GI motility were investigated by calculating percent intestinal transit rates (ITR). Results : 1. The ICC generated pacemaker potentials in the murine small intestine. Yijin-tang produced membrane depolarization with concentration-dependent manners in the current clamp mode. 2. Pretreatment with a Ca²⁺ free solution and thapsigargin, a Ca²⁺-ATPase inhibitor in the endoplasmic reticulum, stopped the pacemaker potentials. In the case of Ca²⁺-free solutions and thapsigargin, Yijin-tang did not induce membrane potential depolarizations. 3. U73122, a phospholipase C (PLC) inhibitors, blocked the Yijin-tang-induced membrane potential depolarizations. However, U73343, an inactive PLC inhibitors, did not block. 4. In the presence of protein kinase C (PKC) inhibitors, staurosporine or Rottlerin, Yijin-tang depolarized the pacemaker potentials. However, in the presence of Go6976, Yijin-tang did not depolarize the pacemaker potentials. 5. In mice, intestinal transit rate (ITR) values were significantly and dose-dependently increased by the intragastric administration of Yijin-tang. Conclusions : These results suggest that Yijin-tang can modulate the pacemaker activity of ICC through an internal/external Ca²⁺ and PLC/PKC-dependent pathway in ICC. In addition, Yijin-tang is a good candidate for the development of a prokinetic agent.

• 생명과학회지
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• 제18권9호
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• pp.1194-1201
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• 2008

본 연구는 유해성 Cochlodinium polykrikoides 적조생물을 대상으로 수온변화에 따른 세포 생화학적 및 생리 활성도를 측정했다. Genomic DNA 함량은 $12^{\circ}C$ 및 $15^{\circ}C$에서 거의 비슷한 0.6을 보였으나, $18^{\circ}C$부터 급격히 높아져서 $24^{\circ}C$ 최고 1.8를 나타내었다. RNA와 total protein도 $24^{\circ}C$에 가장 높은 1.7과 0.07 ${\mu}g$ $ml^{-1}$으로 나타났다. 광합성량도 수온에 따른 큰 변화를 보였다. 빛의 파장에 관계없이 $18^{\circ}C$ 이상에서 현저히 높은 값을 보였다. $24^{\circ}C$ $ETR_{max}$ Ch1-Ch4까지의 범위는 537.9에서 602.5 ${\mu}mol$ electrons $g^{-1}$ Chl ${\alpha}s^{-1}$ 나타났다. Nitrate reductase와 ATPase 효소 활성도는 $24^{\circ}C$에서 각각 0.11 ${\mu}mol$ $NO_{2}^{-}$ ${\mu}g^{-1}$ Chl ${\alpha}h^{-1}$ , 0.78 pmol 100 $mg^{-1}$ 나타났다. CHN 분석에서도 수온에 따라 C, H, N의 함량이 현저하게 상이했다. $27^{\circ}C$ 배양시 $24^{\circ}C$에 비하여 대부분의 세포생리물질이 낮게 보였다. 따라서 C. polykrikoides는 수온 변화에 대하여 세포대사물질의 함량이 많은 차이를 볼 수 있어서 초기 적조 발생 조건은 $18^{\circ}C$로 추측된다. 본 실험의 결과로 $24^{\circ}C$ 이상이 되면 C. polykrikoides 대번식은 세포 내 생리물질의 현저한 저하로 형성되기가 어려울 것으로 보인다.