• 제목/요약/키워드: NADPH-d

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인삼약침이 소음스트레스로 인한 태아쥐의 NOS 신경세포 발현에 미치는 영향 (Effect of Ginseng radix herb-acupuncture on noise stress-induced NOS expression in the offspring rats)

  • 이익재;임강현;서일복;구성태;최선미;김이화
    • Korean Journal of Acupuncture
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    • 제23권4호
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    • pp.157-167
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    • 2006
  • 목적 : 인삼은 전통적으로 학습과 기억능력 증진 및 각종 질병치료에 우수한 효과가 있는 것으로 알려져 있다. 본 연구에서는 이러한 인삼약침이 소음스트레스가 유발된 태아쥐의 해마 및 시상하부에서 nitric oxide synthase에 미치는 영향을 관찰하였다. 방법 : 인삼약침이 소음스트레스에 미치는 영향을 연구하기 위하여 태아기에 소음스트레스를 유발하고 생후 4주 후 7일간 인삼약침을 투여한 후 NADPH-d 조직화학법을 시행하였다. 결과 : 소음스트레스는 태아쥐의 해마 및 시상하부에서 NOS 발현이 유의하게 증가하였다. 2. 인삼약침은 해마 CA1에서만 소음스트레스에 의해서 증가된 NOS 발현을 유의하게 억제하였다 3. 인삼약침은 시상 하부 PVN, DMH 및 LHA에서 소음스트레스에 의해서 증가된 NOS 발현을 유의하게 억제하였다. 결론 : 본 연구를 통하여 인삼약침이 소음스트레스가 유발된 태아쥐의 해마 및 시상하부에서 증가된 NOS 발현을 유의하게 억제시켜 태아의 스트레스 자극에 인삼약침치료가 유의한 효과가 있음을 확인하였다.

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Acinetobacter sp. T5-7에 의한 Phenol과 Trichloroethylene 분해특성 (Characterization of Trichloroethylene and Phenol Degradation by Acinetobaeter sp. T5-7)

  • 홍성용;이숙희;이정해;하지홍
    • 한국미생물·생명공학회지
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    • 제23권3호
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    • pp.255-262
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    • 1995
  • Intact cells of Acinetobacter sp. T5-7 completely degraded trichloroethylene (TCE) following growth with phenol. This strain could grow on at least eleven aromatic compounds, e.g., benzaldehyde, benzene, benzoate, benzylalochol, catechol, caffeic acid, 2.4-D, p-hydroxybenzoate, phenol, protocatechuate and salicylate, and did grow on alkane, such as octane. But except phenol, other aromatic compounds did not induced TCE degradation. Phenol biotransformation products, catechol was identified in the culture media. However, catechol-induced cells did not degrade TCE. So we assumed that phenol hydroxylase was responsible for the degradation of TCE. The isolate T5-7 showed growth in MM2 medium containing sodium lactate and catechol rather than phenol, but did not display phenol hydroxyalse activity, suggesting induction of enzyme synthesis by phenol. Phenol hydroxylase activity was independent of added NADH and flavin adenine dinucleotide but was dependent on NADPH addition. Degradation of phenol produced catechols which are then cleaved by meta-fission. We identified catechol-2.3-dioxygenase by active staining of polyacrylamide gel.

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알데히드 옥시다제의 활성에 미치는 리도카인 및 프로카인아미드의 영향 (The Effect of Lidocaine and Procainamide on the Hepatic Aldehyde Oxidase Activity)

  • 허근;김진숙;김대경;하은필;이상일;용철순
    • 약학회지
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    • 제43권6호
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    • pp.756-761
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    • 1999
  • Lipid peroxidation mediated by hydroxyl radicals which are generated during myocardial ischermia has suggested as a possible mechanism of ischemic myocardial damage. Recently, it has been reported that anti-arrhythmic action of lidocaine, a local anesthetic, is attributed to its "membrane-stabilizing" properties through scavenging free radicals, thus, inhibiting lipid peroxidation. Aldehyde oxidase and xanthine oxidase which catalyze the oxidation of many purine, pyrimidine and pteridine derivatives are known as free radical generating systems. In this experiment, we studied the effect of lidocaine and procainamide on the hepatic aldehyde and xanthine oxidase activity and antioxidative activities. It was found that lidocaine and procainamide inhibited both NADPH-dependent and independent lipid peroxidation. Both of tested compounds were found to be ineffective in inhibiting xanthine oxidase. Lidocaine and procainamide, however, inhibited aldehyde oxidase activity in vitro as well as in vivo. Based on the above results, lidocaine and procainamide could be employed as a therapeutic agent for aldehyde oxidaserelated disease.d disease.

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Hydrogen Evolution from Biological Protein Photosystem I and Semiconductor BiVO4 Driven by Z-Schematic Electron Transfer

  • Shin, Seonae;Kim, Younghye;Nam, Ki Tae
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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    • pp.251.2-251.2
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    • 2013
  • Natural photosynthesis utilizes two proteins, photosystem I and photosystem II, to efficiently oxidize water and reduce NADP+ to NADPH. Artificial photosynthesis which mimics this process achieve water splitting through a two-step Z-schematic water splitting process using man-made synthetic materials for hydrogen fuel production. In this study, Z-scheme system was achieved from the hybrid materials which composed of hydrogen production part as photosystem I protein and water oxidizing part as semiconductor BiVO4. Utilizing photosystem I as the hydrogen evolving part overcomes the problems of existing hydrogen evolving p-type semiconductors such as water instability, expensive cost, few available choices and poor red light (>600 nm) absorbance. Some problems of photosystem II, oxygen evolving part of natural photosynthesis, such as demanding isolation process and D1 photo-damage can also be solved by utilizing BiVO4 as the oxygen evolving part. Preceding research has not suggested any protein-inorganic-hybrid Z-scheme composed of both materials from natural photosynthesis and artificial photosynthesis. In this study, to realize this Z-schematic electron transfer, diffusion step of electron carrier, which usually degrades natural photosynthesis efficiency, was eliminated. Instead, BiVO4 and Pt-photosystem I were all linked together by the mediator gold. Synthesized all-solid-state hybrid materials show enhanced hydrogen evolution ability directly from water when illuminated with visible light.

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Evaluation of Brain Damage Induced by Transient Occlusion of Middle Cerebral Artery in Rats

  • Insook Jung;Lee, Juseon;Melissa D. Oh;Kim, Myungsoo;Changbae Jin
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1997년도 춘계학술대회
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    • pp.103-103
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    • 1997
  • 일시적인 국소 뇌허혈 rat model에서 중대뇌동맥의 폐쇄시간에 따른 뇌조직의 손상정도를 비교 조사하였다. 웅성 Wistar rats를 isoflurane 홉입마취하에서 우측 내경동맥내로 17 mm의 silicone-coated 4-0 nylon monofilament를 삽입하여, 중대뇌동맥의 기저부를 30분, 60분 또는 90분동안 폐쇄한 후 이 monofilament를 다시 밖으로 뽑아내므로써 23 시간동안 recirculation 시키는 일시적 국소 뇌허혈 rat model을 사용한 결과, 수술 후 거의 모든 rats에서 left hemiparesis 또는 좌측으로의 circling과 같은 신경학적 결손이 나타나므로써 높은 성공률을 가지고서 비교적 용이하게 뇌허혈을 유발시킬 수 있었으며, 2 mm 두께의 fresh brain coronal slices에 대하여 2,3,5-triphenyltetrazolium chloride (TTC) 염색법을 시행하여 slice surface의 사진을 찍고 computerized 영상분석법을 이용하여 필요한 면적을 측정하므로써, coronal slice의 infarction size (%)는 총 면적에 대한 infarction 면적의 %로서, 부종율 (%)은 대조측 대뇌반구 면적의 2배에 대한 동측 대뇌반구와 대조측 대뇌반구 면적 차이의 %로서 산정되어졌다. 30분 허혈군에서는 본 염색법으로는 infarction이 거의 확인되지 않았으나 60분 허혈군 (n=13)에서는 우측 somatosensory frontoparietal cortex와 striatum 양자 모두 또는 일부 rats에서는 striatum에만 국한된 ulfarction이 확인되어졌으며 90분 허혈군 (n=12)에서는 거의 대부분의 rats에서 위 두 지역 모두에서 infarction이 확인되어졌다. Infarction size (%)는 60분과 90분 허혈군 각각에서, frontal pole로부터 5 mm되는 slice에서는 11.9$\pm$1.2 (평균치$\pm$표준오차), 13.7$\pm$1.9이었으며 7 mm되는 slice에서는 19.1$\pm$1.8, 21.9$\pm$2.1이었으며 9 mm되는 slice에서는 14.7$\pm$2.4, 19.7$\pm$2.2이었다. 또한 부종율 (%)은 60분과 90분 허혈군 각각에서, frontal pole로부터 5 mm되는 slice에서는 3.1$\pm$0.6, 3.8$\pm$0.7이었으며 7 mm되는 slice에서는 3.5$\pm$0.3, 5.7$\pm$1.0이었으며 9 $\pm$되는 slice에서는 3.4$\pm$0.5, 5.7$\pm$0.9이었다. 한편 90분 동안의 중대뇌동맥 폐쇄에 따른 뇌조직 손상을 cresyl violet 염색법, NADPH-diaphorase histochemistry, GFAP immunohistochemistry를 사용하여 일부 측정한 결과, 위의 손상지역에서는 뇌신경세포체들의 shrinkage 내지는 소실됨을 확인할 수 있었으며, NADPH-diaphorase-positive neuron들도 대부분 dendrite와 axon같은 cell process들의 fragmentation, shrinkage 내지는 소실되므로써 intensity의 감소현상이 나타났으며, reactive gliosis로 말미암아 GFAP immunoreactive intensity의 증가현상이 나타났다.

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꽃뱀과 흰쥐의 간 마이크로좀에 존재하는 Cytochrome P45O 의존성 Monooxygenases의 특성 비교 (Comparison of Characteristics of Hepatic Microsomal Cytochrome P45O-dependent Monooxygenases from Snake and Rat)

  • Ja Young Moon;Dong Wook Lee;Ki Hyun Park
    • 생명과학회지
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    • 제8권6호
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    • pp.695-701
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    • 1998
  • 한국산 꽃뱀(Natrix tigrina Lateralis)의 간 마이크로좀에 존재하는 mixed function oxidase system 구성 성분들의 함량과 P45O 의존성 monooxygenase의 활성도를 조사하고 이들을 흰쥐(Sp. D)의 것과 상호 비교하였다. 꽃뱀에서의 P45O, b5 함량 및 NADPH-cytochrome c reductase 활성도는 흰쥐에서 보다 낮았으며, 7-ethoxycoumarin O-deethylase와 benzphetamine N-demethylase 활성도 역시 흰쥐에서 보다 상당히 낮았다. 그러나 aryl hydrocarbon hydroxylase와 testosterone hydroxylase 활성도는 흰쥐와 비교할 때 거의 비슷하거나 오히려 높았다. Testosterone의 수산화 반응에 대한 선택특이성을 조사한 결과, 꽃 뱀은 7$\alpha$ 위치에서, 흰쥐는 6$\beta$ 위치에서 가장 높은 수산화 반응물을 생성했다. 그러나 testosterone의 C2와 C3 위치에서의 수산화 반응에 대한 선택특이성은 꽃뱀과 흰쥐에서 비슷하였다. Radioimmunoassay (RlA)를 실시하여 5종 (CYP2B, CYP1A1, CYP1A2, CYP3A 및 CYP2El)의 P45O 동위효소의 구성비를 비교한 결과, 꽃뱀에서는 CYP1A1/1A2가, 흰쥐에서는 CYP2El이 각각 비교적 많이 존재하였다. 부분정제한 P45O을 SDS-PAGE와 RIA로 분석한 결과, 꽃뱀의 간 마이크로좀에 존재하는 P45O중에는 흰쥐와는 다른 종류의 P45O 동위효소가 존재함을 시사하였다.

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The Protective Effect of Quercetin-3-O-${\beta}$-D-Glucuronopyranoside on Ethanol-induced Damage in Cultured Feline Esophageal Epithelial Cells

  • Cho, Jung-Hyun;Park, Sun-Young;Lee, Ho-Sung;Whang, Wan-Kyunn;Sohn, Uy-Dong
    • The Korean Journal of Physiology and Pharmacology
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    • 제15권6호
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    • pp.319-326
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    • 2011
  • Quercetin-3-O-${\beta}$-D-glucuronopyranoside (QGC) is a flavonoid glucoside extracted from Rumex Aquaticus Herba. We aimed to explore its protective effect against ethanol-induced cell damage and the mechanism involved in the effect in feline esophageal epithelial cells (EEC). Cell viability was tested and 2',7'-dichlorofluorescin diacetate assay was used to detect intracellular $H_2O_2$ production. Western blotting analysis was performed to investigate MAPK activation and interleukin 6 (IL-6) expression. Exposure of cells to 10% ethanol time-dependently decreased cell viability. Notably, exposure to ethanol for 30 min decreased cell viability to 43.4%. When cells were incubated with $50{\mu}M$ QGC for 12 h prior to and during ethanol treatment, cell viability was increased to 65%. QGC also inhibited the $H_2O_2$ production and activation of ERK 1/2 induced by ethanol. Pretreatment of cells with the NADPH oxidase inhibitor, diphenylene iodonium, also inhibited the ethanol-induced ERK 1/2 activation. Treatment of cells with ethanol for 30 or 60 min in the absence or presence of QGC exhibited no changes in the IL-6 expression or release compared to control. Taken together, the data indicate that the cytoprotective effect of QGC against ethanol-induced cell damage may involve inhibition of ROS generation and downstream activation of the ERK 1/2 in feline EEC.

Histamine Release by Hydrochloric Acid is Mediated via Reactive Oxygen Species Generation and Phospholipase D in RBL-2H3 Mast Cells

  • Kim, Chang-Jong;Lee, Seung-Jun;Seo, Moo-Hyun;Cho, Nam-Young;Sohn, Uy-Dong;Lee, Moo-Yeol;Shin, Yong-Kyoo;Sim, Sang-Soo
    • Archives of Pharmacal Research
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    • 제25권5호
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    • pp.675-680
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    • 2002
  • In order to investigate the underlying mechanism of HCI in oesophagitis, the inflammatory response to HCI was observed in RBL-2H3 mast cells. Rat basophilic leukemia (RBL-2H3) cells were used to measure histamine release, arachidonic acid (AA) release, reactive oxygen species (ROS) and peroxynitrite generation induced by HCI. Exogenous HCl increased the level of histamine release and ROS generation in a dose dependent manner, whereas it decreased the spontaneous release of [$^3$H] M and the spontaneous production of peroxynitrite. Mepacrine (10 $\mu$M), oleyloxyethyl phosphorylcholine (10 $\mu$M) and bromoenol lactone (10 $\mu$M) did not affect both the level of histamine release and ROS generation induced by HCI. U73122 (1 $\mu$M), a specific phospholipase C (PLC) inhibitor did not have any influence on level of histamine release and ROS generation. Propranolol (200 $\mu$M), a phospholipase D (PLD) inhibitor, and neomycin (1 mM), a nonspecific PLC and PLD inhibitor, significantly inhibited both histamine release and ROS generation. Diphenyleneiodonium (10 $\mu$M), a NADPH oxidase inhibitor, and tiron (5 mM), an intracellular ROS scavenger significantly inhibited the HCI-induced histamine release and ROS generation. These findings suggest that the inflammatory responses to HCI is related to histamine release and ROS generation, and that the ROS generation by HCI may be involved in histamine release via the PLD pathway in RBL-2H3 cells.

Pyruvate Protection against Endothelial Cytotoxicity Induced by Blockade of Glucose Uptake

  • Chung, Se-Jin;Lee, Se-Hee;Lee, Yong-Jin;Park, Hyoung-Sook;Bunger, Rolf;Kang, Young-Hee
    • BMB Reports
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    • 제37권2호
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    • pp.239-245
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    • 2004
  • We have previously demonstrated that the redox reactant pyruvate prevents apoptosis in the oxidant model of bovine pulmonary artery endothelial cells (BPAEC), and that the anti-apoptotic mechanism of pyruvate is mediated in part via the mitochondrial matrix compartment. However, cytosolic mechanisms for the cytoprotective feature of pyruvate remain to be elucidated. This study investigated the pyruvate protection against endothelial cytotoxicity when the glycolysis inhibitor 2-deoxy-D-glucose (2DG) was applied to BPAEC. Millimolar 2DG blocked the cellular glucose uptake in a concentration- and time-dependent manner with >85% inhibition at $\geq$5 mM within 24 h. The addition of 2DG evoked BPAEC cytotoxicity with a substantial increase in lipid peroxidation and a marked decrease in intracellular total glutathione. Exogenous pyruvate partially prevented the 2DG-induced cell damage with increasing viability of BPAEC by 25-30%, and the total glutathione was also modestly increased. In contrast, 10 mM L-lactate, as a cytosolic reductant, had no effect on the cytotoxicity and lipid peroxidation that are evoked by 2DG. These results suggest that 2DG toxicity may be a consequence of the diminished potential of glutathione antioxidant, which was partially restored by exogenous pyruvate but not L-lactate. Therefore, pyruvate qualifies as a cytoprotective agent for strategies that attenuate the metabolic dysfunction of the endothelium, and cellular glucose oxidation is required for the functioning of the cytosolic glutathione/NADPH redox system.

Salmonella typhimurium에서 유래한 Mannitol Dehydrogenase 유전자의 대장균 내 발현 및 효소특성 규명 (Enzymatic Characterization of Salmonella typhimurium Mannitol Dehydrogenase Expressed in Escherichia coli)

  • 장명운;박정미;김민정;강정현;이소원;김태집
    • 미생물학회지
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    • 제48권2호
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    • pp.156-162
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    • 2012
  • Salmonella typhimurium LT2 (KCTC 2421)로부터 mannitol dehydrogenase (StMDH)로 추정되는 유전자를 클로닝하고, 대장균에서 대량 발현하였다. 이 유전자는 488개의 아미노산 서열(약 54 kDa)을 암호화하는 1,467 bp의 염기로 구성되며, 이미 보고된 long-chain dehydrogenase/ reductase (LDR) 계열 효소들과 약 36%의 아미노산 서열 상동성을 나타내었다. 재조합 StMDH의 최적 반응온도는 $30^{\circ}C$이며, pH 5.0의 조건에서 최대의 D-fructose 환원활성, 그리고 pH 10.0에서 최대의 D-mannitol 산화활성을 보인다. 반면에 glucose, galactose, xylose, arabinose 등의 기질에 대해서는 활성을 보이지 않았다. 이 효소는 $NAD^+$/NADH 존재 하에서만 산화 환원 활성을 가지며, $NADP^+$/NADPH는 조효소로 이용하지 못하였다. 결론적으로 StMDH는 전형적인 $NAD^+$/NADH 의존형의 mannitol dehydrogenase (EC 1.1.1.67)임을 확인하였다.