• Title/Summary/Keyword: cytosol

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NOS(Nitric Oxide Synthase) 작용 검색법

  • 이종화
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.151-151
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    • 1993
  • A. 1. 중추신경계에서 일어나는 병변들, Alzheimer's disease, Parkinsonism 및 정상적인 노화현상들을 구명하는 연구의 일부분으로 뇌내의 신경세포들의 세포내물질의 변화를 관찰하고자, 이 실험을 시도하였다. 2. 중추신경계에 작용하는 각종 약물들의 작용기전, 그리고 부작용을 구명하여 그의 예방 및 치료방법을 모색하고자 한다. B. 1.Preparations: rat brain cytosol & culture cell-line(NIE-115) 2. Method: NO or NOS의 생성함량을 직접측정 할 수 없으므로 간접방법을 택한다. [3Hlarginine 및 [3Hlguanine을 Preparation에 incubation시켜 일정한 시간 후 생성되는 [3H]citrulline 또는 cyclic GMP을 scintillation counter로 측정하여 그 함량으로 NO or NOS의 생성을 측정한다.

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Antidiabetic activity of Cultivated Cordyceps pruinosa

  • Kim, Tae-Woong;Sung, Jae-Mo;Yang, Ki-Sook
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.209.3-209.3
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    • 2003
  • Cordyceps species has been used as antiinflammatory, antitoxic, diuretic in folk remedies. Recent research has been reported the effect of anticancer, antidiabetic, antimutagenic, antilipid peroxidation. We examined the antidiabetic activity of cultivated Cordyceps pruinosa on streptozotocin-induced diabetic rats. The blood glucose level was recovered by treatment with Cordyceps pruinosa ethanol extract. The contents of GPT, total cholesterol and xanthine oxidase, glutathione peroxidase, catalase activities of the cytosol were significantly decreased as compared to the diabetic group. (omitted)

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Antimutagenic Activities of Cell Wall and Cytosol Fractions of Lactic Acid Bacteria Isolated from Kimchi

  • Park, Kun-Young;Kim, So-Hee;Son, Tae-Jin
    • Preventive Nutrition and Food Science
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    • v.3 no.4
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    • pp.329-333
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    • 1998
  • Cell wall (lactic acid bacteria-sonicated precipitate ; LAB-SP) and cytosoll(lactic acid bacteria-sonicated supernatant ; LAB-SS) fractions were prepared from kimchi fermenting lactic acid bacteria such as Leuconostoc mesenteroides, Lactobacillus brevis, Lactobacillus fermentum , Lactobacillus plantarum and Pediococcus acidilactici, with Lactobacillus acidophillus isolated from yogurt. Using the Ames mutagenicity test and SOS chormotest system, the antimutagenic acitivity of those cell fractions was studied . One hundered eighty $\mu$l of LAB-SP from lactic acid bacteria isolated from kimchi, excepting Pediococcus acidilactici, supressed the mutagenicity of 4-nitroquinoline-1-oxide(4-NQO) in Ames mutagenicity test and SOS chromotes system , by above 90% and 60% , respectively. LAB-SP from lactic acid bacteria also inhibited the mutagenicity mediated by 3-amino-1-methyl-5H-pyrido [4,3-b]indole (Trp-P-2). Lactobacillus fermentum, Lactobacillus plantarum, and Lactobacillus acidphillus had higher antimutagenicity against Trp-P-2). Lactobacillus fermentum , Lactobacillus plantarum , and Lactobacillus acidphillus had higher antimutagenicity against Trp-P-2 than the other lactic acid bacteria. However, LAB-SS of lactic acid bacteria did not show any mutagenic activity against 4-NQO in Ames mutagenicity test and SOS chromotest systems. On the mutagenicity of MEIQ and Trp-P-2 , LAB-SS of lactic acid bacteria from kimchi or dairy products exhibited a weaker inhibitory effect than LAB-SP of those bacteria. These results represent that, whether the lactic acid bacteria from kimchi are viable or nonviable, antimutagenic acitivity was still effective. We suggest that the strong, antimutaganic activity of lactic acid bacteria might be found in the cell wall fraction , rather than in the cytosol fraction.

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Delivery of Chicken Egg Ovalbumin to Dendritic Cells by Listeriolysin O-Secreting Vegetative Bacillus subtilis

  • Roeske, Katarzyna;Stachowiak, Radoslaw;Jagielski, Tomasz;Kaminski, Michal;Bielecki, Jacek
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.122-135
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    • 2018
  • Listeriolysin O (LLO), one of the most immunogenic proteins of Listeria monocytogenes and its main virulence factor, mediates bacterial escape from the phagosome of the infected cell. Thus, its expression in a nonpathogenic bacterial host may enable effective delivery of heterologous antigens to the host cell cytosol and lead to their processing predominantly through the cytosolic MHC class I presentation pathway. The aim of this project was to characterize the delivery of a model antigen, chicken egg ovalbumin (OVA), to the cytosol of dendritic cells by recombinant Bacillus subtilis vegetative cells expressing LLO. Our work indicated that LLO produced by non-sporulating vegetative bacteria was able to support OVA epitope presentation by MHC I molecules on the surface of antigen presenting cells and consequently influence OVA-specific cytotoxic T cell activation. Additionally, it was proven that the genetic context of the epitope sequence is of great importance, as only the native full-sequence OVA fused to the N-terminal fragment of LLO was sufficient for effective epitope delivery and activation of $CD8^+$ lymphocytes. These results demonstrate the necessity for further verification of the fusion antigen potency of enhancing the MHC I presentation, and they prove that LLO-producing B. subtilis may represent a novel and attractive candidate for a vaccine vector.

Effect of Hovenia dulcis THUNBER var. koreana Nakai Fruits Extracts on Glucose, Lipid Metabolism and Antioxidant Activities in Streptozotocin Induced Diabetic Rat

  • Lee Yoon-Ah;Chae Hee-Jun;Moon Hae-Yeon
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.533-538
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    • 2005
  • This investigation was performed to study the antioxidant activities of Hovenia dulcis THUNBER var. koreana Nakai fruits extracts and the effect of Hovenia dulcis fruits extracts on glucose, lipid metabolism in diabetic rats. DPPH free radical scavanging activitiy and superoxide anion radical Scavenging of Hovenia dulcis fruits $80\%$ methanol extracts were $0.06\pm0.002mg$ polyphrnol/ml and $0.l2\pm0.00lmg$ polyphmol/ml, respectively. Hovenia dulcis fruits $80\%$ methanol extracts were partitioned into hexan, dichloro methane, ethyl acetate and butanol, successively. Ethyl acetate fraction were good antioxidant activity. Streptozotocin (45 mg/kg body weight, i.p.) induced diabetic rats showed a significant increases of plasma glucose, triglyceride and total cholesterol. Concomitantly significant decrease of plasma high density lipoprotein. Glutathione level were decrease in cytosol of liver. Lipid peroxide were increase in microsome of liver. Group 1 and 2 were treated with Hovenia dulcis fruits ethyl acetate extracts 50 mg/kg body weight and 20mg/kg body weight for 24 days, individually. Group land 2 rats showed decreased plasma glucose, triglyceride, total cholesterol and lipide peroxide in microsome of liver tissue of rats, and increased plasma high density lipoprotein and glutathione in cytosol of liver tissue rats. The result suggest that Hovenia dulcis THUNBER var. koreana Nakai fruits extracts may effectively normalize the impaired antioxiants status in streptozotocin induced diabetic rats. Hovenia dulcis fruits ethyl acetate extracts were used to improve the imbalance between free radicals and antioxidant system due to the diabetes.

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Studies on the Membrane Topology of the (Na, K) ATPase

  • Lee, Kyunglim-Yoon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.181-181
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    • 1996
  • The (Na, K) ATPase is a membrane ion transporting ATPase composed of an ${\alpha}$ catalytic subunit and a ${\beta}$ glycoprotein subunit. The topology of the rat ${\alpha}$1 and ${\beta}$1 subunits has been studied by insertion of epitope(s) : at the NH2-terminus and COOH-terminus and between Glu117 and Glul18, Lys828 and Arg829, Gln900 and Trp901, and Va1939 and Phe940 of the ${\alpha}$ subunit; and at the NH2-terminus and COOH-terminus and between Glu228 and Tyr229 of the ${\beta}$ subunit. The epitope-tagged ${\alpha}$l, constructs were expressed in HeLa cells to select for stable cell lines expressing a functional (Na, K)ATPase. All constructs, except for the one tagged between Gln900 and Trp901, resulted in ouabain-resistant colonies indicating that modified proteins retained functional integrity. The epitope-tagged ${\beta}$ constructs were transiently expressed in Cos-7 cells. The orientation of the epitopes with respect to the cell membrane was revealed by indirect immunofluorescence performed on permeabilized and non-permeabilized cells expressing the (Na, K)ATPase chains. The results indicate that the ${\alpha}$ subunit has 4 transmembrane segments in the COOH terminal membrane bound domain between residues 760 and 938, and that both the NH2-terminus and the COOH-terminus are in the cytosol; it was not determined whether there are more transmembrane segments between residue 938 and the COOH-terminus. The ${\beta}$ subunit has only one transmembrane spanning region with the NH2-terminus in the cytosol and the COOH-terminus on the extracytoplasmic surface of the plasma membrane.

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Induction of the Intrinsic Apoptotic Pathway by 3-Deazaadenosine Is Mediated by BAX Activation in HL-60 Cells

  • Lee, Sun-Young;Ko, Kyoung-Won;Kang, Won-Kyung;Choe, Yun-Jeong;Kim, Yoon-Hyoung;Kim, In-Kyung;Kim, Jin;Kim, Ho-Shik
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.6
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    • pp.407-412
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    • 2010
  • 3-Deazaadenosine (DZA), a potent inhibitor of S-adenosylhomocysteine hydrolase, was previously proposed to induce intrinsic apoptosis in human leukemic cells. In the present study, we analyzed the mechanism underlying the DZA-induced intrinsic apoptotic pathway. DZA activated typical caspase-dependent apoptosis in HL-60 cells, as demonstrated by an accumulation of hypo-diploidic cells, the processing of multiple procaspases and an inhibitory effect of z-VAD-Fmk on this cell death. During DZA-induced apoptosis, cytochrome c (cyt c) was released into the cytosol. This was neither prevented by z-VAD-Fmk and nor was it associated with the dissipation of mitochondrial membrane potential (${\Delta}{\Psi}_m$). Prior to the release of cyt c, BAX was translocated from the cytosol to mitochondria and underwent oligomerization. Finally, the overexpression of BCL-XL protected HL-60 cells from apoptosis by blocking both the cyt c release and BAX oligomerization. Collectively, these findings suggest that DZA may activate intrinsic apoptosis by stimulating BAX activation and thereby the release of cyt c.

Reduction of Muscarinic $K^+$ Channel Activity by Transferrin in Ischemic Rat Atrial Myocytes

  • Park, Kyeong-Tae;Kang, Da-Won;Han, Jae-Hee;Hur, Chang-Gi;Hong, Seong-Geun
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.6
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    • pp.333-339
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    • 2003
  • It has been demonstrated that an unidentified cytosolic factor(s) reduces $K_{ACh}$ channel function. Therefore, this study attempted to elucidate the cytosolic factor. Fresh cytosol isolated from normal heart (FC) depressed the $K_{ACh}$ channel activity, but cytosol isolated from the ischemic hearts (IC) did not modulate the channel function. Electrophorectic analysis revealed that a protein of ${\sim}80 kDa was markedly reduced or even lost in IC. By using peptide sequencing analysis and Western blot, this 80 kDa protein was identified as transferrin (receptor-mediated $Fe^{3+}$ transporter, 76 kDa). Direct application of transferrin (100 nM) to the cytoplasmic side of inside-out patches decreased the open probability ($P_o$, 12.7${\pm}6.4%, n=4) without change in mean open time (${\tau}_o$, $98.5{\pm}1.3$%, n=4). However, the equimolar apotransferrin, which is free of $Fe^{3+}$, had no effect on the channel activity (N*$P_o$, $129.1{\pm}13.5$%, n=3). Directly applied $Fe^{3+}$ (100 nM) showed results similar to those of transferrin (N*$P_o$: $21.1{\pm}3.9$%, n=5). However $Fe^{2+}$ failed to reduce the channel function (N*$P_o$, $106.3{\pm}26.8$%, n=5). Interestingly, trivalent cation La3+ inhibited N*$P_o$ of the channel ($6.1{\pm}3.0$%, n=3). Taken together, these results suggest that $Fe^{3+}$ bound to transferrin can modulate the $K_{ACh}$ channel function by its electrical property as a polyvalent cation.

Evolutionary Study on the Dark Chub (Zacco temmincki) 1. Geographic Distribution and Seasonal Variation of Two Allelomorphs of MDH (갈겨니(Zacco temmincki)의 진화에 관한 연구 1. MDH 동위효소 2형의 지리적 분포 및 계절적 변이)

  • 양서영;민미숙
    • The Korean Journal of Zoology
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    • v.32 no.3
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    • pp.232-241
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    • 1989
  • Two cytosol malate dehydrogenase allelomorphs are found in the Dark Chub, Zacco temmincki, a freshwater fish inhabiting Korean waters. Mdh-1MM(hereafter referred to as A-type) is distributed along the south and southeastern parts of Korea, whereas the B-type(Mdh-1MS) is found everywhere except the southeast. Several syrnpatric populations can be found in the southern areas with no hybridization between them. The pattern of geographic distribution would indicate that A-type is superior adaptation to colder water temperature; this is collaborated by evidence collected from a sympatric length of the Tongchon stream, Namhae. The relative warmth of the water along this portion of the stream's length was determined its altitude and the time of year. Frequency of B-types found was directly proportional to the relative warmth of the water and inversely proportional to the nurnber of A-types found in the sarne station. Current data tends to suggest that the B-type of Zacco temrnincki first differentiated from Zacco platypus approximately one million years ago (Yang et al., 1984) while A-type subsequently branched from B-type 300,000 years later(Yang and Mm, 1987) through a progressive adaptation toward colder temperature.

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Effect of the Toluene Administration on the Liver Damage in Rats Fed Low or Standard Protein Diet (저 및 표준단백식이로 성장시킨 흰쥐에 Toluene 투여가 간 손상에 미치는 영향)

  • 윤종국;김경순;임영숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.6
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    • pp.981-985
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    • 1996
  • To study the effect of toluene administration on the liver damage, rats were previously fed a low (casein 7%, LP) or standard(casein 20%, SP) protein diet and for four days toluene(50% in olive oil) was given at 0.2ml/100g body weight/day to the male rats, and then the degree of liver damage in toluenetreated animals fed LP were compared with those fed SP. The increasing rate of liver weight/body weight and the serum levels of xanthine oxidase to the control group were higher in rats fed SP than those fed LP. The decreasing rate of protein contents in cytosol, mitochondria and glycogen, glutathione contents of liver to the control group were higher in rats fed SP than those fed LP. In histopathological findings, the swelling of hepatic cell around the central vein was demonstrated in all the two groups toluene-treated rats. But the degree of swelling severity in hepatocytes was somewhat higher in rats fed SP than those fed LP. Therefore it is assumed that the degree of liver damage severity in toluenetreated animals was higher in rats fed SP than those fed LP.

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