• Title/Summary/Keyword: Microsomal fraction

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The degradation o Diazinon by hepatic monooxygenase of Pig (돼지 간중의 Monooxygenase 가 Diazinon 의 분해에 미치는 영향)

  • Ryoo, Jong-Gook;Lee, Kyu-Seung
    • Korean Journal of Environmental Agriculture
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    • v.9 no.2
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    • pp.153-159
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    • 1990
  • Two fractions(microsomal and soluble) were prepared by ultracentrifugation(105,000G for 1hr at $4^{\circ}C$) from pig liver in order to find the major factor in Diazinon degradation. The two enzyme activities showed the same value, but Diazinon was degraded three times in microsomal fraction more than in soluble fraction. And with addition of EPN, Beam and PBO, degradation of diazinon was inhibited(29, 30 and 60%) as well as Monooxygenase activity (14, 15 and 35%) in microsomal fraction, respectively.

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The effect of Aconiti tuber butanol fraction on the rabbit heart microsomal $Na^+-K^+$-activated ATPase activity (부자 Butanol Fraction이 가토 심장근 Microsomal $Na^+-K^+$-activated ATPase 활성도에 미치는 영향)

  • Shin, S.G.;Lim, J.K.;Park, C.W.;Kim, M.S.
    • The Korean Journal of Pharmacology
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    • v.12 no.1
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    • pp.7-14
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    • 1976
  • Aconiti tuber butanol fraction shows positive inotropic effect on the isolated atrium of rabbit heart. To investigate the mechanism, the effect on microsomal ATPase activity of rabbit heart is observed. The microsomal fraction which contains the $Na^+$- and $K^+$-activated ATPase in the presence of $Mg^{++}$ is isolated from the left ventricle of rabbit heart. The microsomal ATPase activity is maximally stimulated at $Na^+$ and $K^+$ concentration of 100 mM and 10 mM respectively. Microsomal $Na^+-K^+$-activated ATPase is inhibited by ouabain and Aconiti tuber butanol fraction. Ouabain and Aconiti tuber butanol fraction depress $Na^+$-stimulation on microsomal ATPase activity, and the inhibitory effects are not completely reversed at $Na^+$ concentration of 300 mM. Also, $K^+$-stimulation on microsomal ATPase activity is inhibited by ouabin and Aconiti tuber butanol fraction and the inhibitions are not compeletely reversed at $K^+$ concentration of 30 mM. It is, therefore, suggested that the inhibitory effect of Aconiti tuber butanol fraction on the microsomal ATPase activity may contribute to leading to the positive inotropic effect.

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A Study on the Mechanism of Insulin Sensitivity to Glucose Transport System: Distribution of Subcellular Fractions and Cytochalasin B Binding Proteins (인슐린의 포도당 이동 촉진 기전에 관한 연구 -세포내부 미세구조와 Cytochalasin B 결합단백질의 분포-)

  • Hah, Jong-Sik
    • The Korean Journal of Physiology
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    • v.24 no.2
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    • pp.331-344
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    • 1990
  • What makes glucose transport function sensitive to insulin in one cell type such as adipocyte, and insensitive in another such as liver cells is unresolved question at this time. Recently it is known that insulin stimulates glucose transport in adipocytes largely by redistributing transporter from the storage pool that is included in a low density microsomal fraction to plasma membrane. Therefore, insulin sensitivity may depend upon the relative distribution of gluscose transporters between the plasma membrane and in an intracellular storage compartment. In hepatocytes, the subcellular distribution of glucose transporter is less well documented. It is thus possible that the apparent insensitivity of the hepatocyte system could be either due to lack of the constitutively maintained, intracellular storage pool of glucose transporter or lack of insulin-mediated transporter translocation mechanism in this cell. In this study, I examined if any intracellular glucose transporter pool exists in hepatocytes and this pool is affected by insulin. The results obtained summarized as followings: 1) Distribution of subcellular fractions of hepatocyte showed that there are $24.9{\pm}1.3%$ of plasma membrane, $36.9{\pm}1.7%$ of nucleus-mitochondria enriched fraction, $23.5{\pm}1.2%$ of lysosomal fraction, $9.6{\pm}1.0%$ of high density microsomal fraction and $4.9{\pm}0.5%$ of low density microsomal fraction. 2) In adipocyte, there were $29.9{\pm}2.6%$ of plasma membrane, $19.4{\pm}1.9%$ of nucleus-mitochondria enriched fraction, $26.7{\pm}1.8%$ of high density microsomal fraction and $23.9{\pm}2.1%$ of low density microsomal fraction. 3) Surface labelling of sodium borohydride revealed that plasma membrane contaminated to lysosomal fraction by $26.8{\pm}2.8%$, high density microsomal fraction by $8.3{\pm}1.3%$ and low density microsomal fraction by $1.7{\pm}0.4%$ respectively. 4) Cytochalasin B bound to all of subcellular fractions with a Kd of $1.0{\times}10^{-6}M$. 5) Photolabelling of cytochalasin B to subcellular fractions occurred on 45 K dalton protein band, a putative glucose transporter and D-glucose inhibited the photolabelling. 6) Insulin didn't affect on the distribution of subcellular fractions and translocation of intracellular glucose transporters of hepatocytes. 7) HEGT reconstituted into hepatocytes was largely associated with plasma membrane and very little was found in low density microsomal fraction which equals to the native glucose transporter distribution. Insulin didn't affect on the distribution of exogeneous glucose transporter in hepatocytes. From the above results it is concluded that insulin insensitivity of hepatocyte may due to lack of intracellular storage pool of glucose transporter and thus intracellular storage pool of glucose transporter is an essential feature of the insulin action.

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The antioxidative effects of α-tocopherol on the lipid peroxidation and protein oxidation by free radicals (Free radical에 의한 지질과산화와 단백질산화에 대한 α-tocopherol의 항산화효과)

  • Chung, Chung-won;Huh, Rhin-sou
    • Korean Journal of Veterinary Research
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    • v.34 no.2
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    • pp.249-258
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    • 1994
  • This study was carried out to investigate the inhibitory effects of vitamin E on the oxidative damage of cellular lipids and proteins in free radical reaction induced by $FeCl_3$, and ascorbic acid. In this experiment, a vitamin E treated rat group was administered with 100mg/kg body weight of $dl-{\alpha}-tocophery$ 1 acetate and an untreated rat group was administered with the same volume of corn oil. And then assays of malondialdehyde and carbonyl group in total homogenate, mitochondrial and microsomal fraction of rat liver were carried out at the scheduled time. The results obtained from this study were summarized as follows; 1. Lipid peroxidation levels in vitamin E administered rat liver cells were significantly (p<0.05) decreased at the intervals between 1 hour and 4 hours in liver homogenate, at all times except for 1 hour point in mitochondrial fraction, and also at the intervals between 0.5 hour and 3 hours in microsomal fraction compared with those of the control rat liver cell. 2. Protein oxidation levels in vitamin E administered rat liver cell were also significantly (p<0.05) decreased at the intervals between 1.5 hours and 4 hours in liver homogenate, at over 4 hours in liver mitochondrial fraction, and at the intervals between 0.5 hour and 3 hours in liver microsomal fraction compared with those of the control rat liver cells.

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Effect of combination treatment of vitamin E and insulin in streptozotocin-treated rats and BB rats I. Effect on the oxidative modification of lipid and protein, and enzyme activitis (BB 랫드 및 streptozotocin이 투여된 랫드에서 vitamin E와 insulin 병합 투여 영향 I. 지질과 단백질의 산화성 손상 및 혈청내 효소 활성도에 미치는 영향)

  • Kim, Soon-tae;Huh, Rhin-sou
    • Korean Journal of Veterinary Research
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    • v.35 no.4
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    • pp.699-712
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    • 1995
  • The present study, to evaluate the effect of vitamin E on the oxidative stress in STZ-treated rat and BB rat, was investigated the biochemical enzyme activity in the serum, and malondialdehyde and carbonyl group in the RBC membrane, liver and microsomal fraction after vitamin E and/ or insulin treatment. Results obtained through the experiments were summarized as follows; 1. Effect of vitamin E and/or insulin treatment in STZ-treated rat 1) Lipid peroxidation level in RBC membrane, liver and microsomal fraction was significantly decreased in vi. tamin E and/or insulin treatment group, and especially more significantly decreased in vitamin E with insulin treated group. 2) Protein oxidation level in RBC membrane, liver and microsomal fraction was significantly decreased in vitamin E and/or insulin treatment group. And it was especially more significantly decreased in RBC membrane and liver of vitamin E with insulin treated group. 3) In the enzyme activity in the serum, the activity of AST and ALT was not altered in all experimental group. The increased ALP activity in STZ-treated group was significantly decreased in insulin treated group and vitamin E with insulin treated group. 4) Decreased level of albumin and creatinine after STZ treatment was significantly increased in vitamin E and/or insulin treated group. 5) Level of glucose, cholesterol and triacylglycerol in serum: Glucose level was not significantly different in vitamin E treated group compared to STZ control group. But it was significantly different in the insulin treated group and vitamin E with insulin treated group compared to STZ control group. The cholesterol content in the serum was significantly increased in STZ control group compared to normal control group. And except low dose vitamin E treatment group, it was significantly decreased in vitamin E and/or insulin treated group compared to STZ control group. The triacylglycerol content in the serum was significantly decreased in STZ control group and increased in high dose vitamin E treated group and vitamin E with insulin treated group. But it was not significantly different in low dose vitamin E treated group and insulin treated group compared to STZ control group. 2. Effect of vitamin E and/or insulin treatment in BB rat 1) Lipid peroxidation level in liver was decreased by vitamin E with insulin treatment compared to insulin treatment. But it was not different in microsomal fractions. 2) Protein oxidation level in liver and microsomal fraction was decreased by vitamin E with insulin treatment compared to insulin treatment only in microsomal fractions. These results suggest that the combination treatment of vitamin E and insulin could prevent the oxidative change of lipid and protein of the RBC membrane, liver and microsomal fraction in STZ-treated rats and BB rats.

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$^3H$-ouabain Binding in Heart Following Infusion of Ouabain in Rabbit (Ouabain 점적투여후 토끼심장에 있어서 $^3H$-Ouabain 결합에 관한 연구)

  • 김상건;김낙두
    • YAKHAK HOEJI
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    • v.30 no.3
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    • pp.149-156
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    • 1986
  • Many experiments have showed that the sodium and potassium ion transporting system and the Na, $^+K^+$-ATPase activity of membrane fragments are inhibited by digitalis glycosides and that the pump may be associated with the pharmacological receptor for the drugs. The aim of our investigation is to elucidate the ouabain binding sites occupation in heart following infusion of ouabain to intact animals by the $^3H$-ouabain binding assay. Lethal dose and 26 percent of lethal dose of ouabain were infused to intact rabbit through ear vein. Microsomal fraction was fractionated from ouabain treated rabbit heart. $^3H$-ouabain binding to these fraction in vitro was studied by the Schwartz's method. $^3H$-ouabain binding to heart microsomal fraction was also studied following infusion of ginseng ethanol extract and caffeine to rabbits respectively. 1) The infusion of lethal dose ouabain (113$\mu\textrm{g}$/kg) inhibited the specific $^3H$-ouabain binding to rabbit heart microsomal fraction to the level of 60% (p<0.01) of control group and the infusion of 26% of lethal dose of ouabain led to the level of 79% (p<0.01) of the control group. 2) Time course of binding of 0.4$\mu{M}$ $^3H$-ouabain to microsomal fraction from rabbit heart following infusion of lethal and 26% of lethal dose of ouabain showed dose dependence at various incubation time. 3) Compared with control, only slight change of $K_d$ and $B_{max}$ was detected in in vitro $^3H$-ouabain binding after infusion of ginseng ethanol extract (300mg/kg) to rabbit. 4) In caffeine infusion group, $^3H$-ouabain binding yielded nearly the same results as control group.

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The Effect of $Mg^#$, $Ca^#$, $Na^+$, $K^+$ and Creatine Phosphate on the ATPase Activity of Microsomal Fraction from Rabbit Uterus (가토자궁근(家兎子宮筋)에서 분리(分離)한 Microsome 분획내(分劃內) Adenosinetriphosphatase 활성도(活性度)에 미치는 $Mg^#$, $Ca^#$, $Na^+$, $K^+$ 및 Creatine phosphate의 영향(影響))

  • Choi, Sin-Jyoung
    • The Korean Journal of Pharmacology
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    • v.2 no.2
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    • pp.35-42
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    • 1966
  • The author investigated the effect of $Mg^#$, $Ca^#$, $Na^+$, $K^+$ and creatine phosphate on the ATPase activity of microsomal fraction isolated from rabbit uterus and obtained the following results : 1) The uterine microsomal fraction contained the $Na^+-$ and $K^+-$ activated ATPase in the presence of $Mg^#$. The ATPase activity increased with protein content in the fraction. 2) The maximum ATPase activity was obtained at $Na^+$ and $K^+$ concentraction of 100 mM respectively. 3) In the absence of $Mg^#$, the ATPase was not activated by $Na^+$ and $K^+$, but inhibited. 4) Car stimulated the $Na^+-$ and $K^+-$ activated ATPase in the presence of $Mg^#$. However, in the absence of $Mg^#$, the ATPase was not activated by $Ca^#$. 5) The $K^+-$ activated ATPase activity was greater than the $Na^+-activated$ ATPase under all conditions. 6) The $Na^+-$ and $K^+$ activated ATPase activity was increased by addition of creatine phosphokinase and creatine phosphate to the reaction mixture.

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Effect of Capsicum Components on Liver Microsomal Cytochrome $P_{450}$ in Rat (고초(苦椒)가 백서(白鼠)의 간(肝) 마이크로좀 Cytochrome $P_{450}$에 미치는 영향(影響))

  • Kong, Young-Ok;Kim, Chang-Soo;Kim, Nak-Doo;Chough, Yun-Sung
    • Korean Journal of Pharmacognosy
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    • v.10 no.1
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    • pp.17-22
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    • 1979
  • The investigation is involved with the effect of Capsicum component on the drug metabolism. To investigate the effects of Capsicum component on, in vivo, drug metabolism in rat, Capsicum acetone extract was given intraperitoneally to mice or rats. The duration of loss of righting reflex was determined as hexobarbital sleeping time in mice. Plasma hexobarbital concentration was also measured by Brodie's method. The rats were pretreated with Capsicum extract acutely or chronically. As the results, hexobarbital sleeping time and plasma hexobarbital concentration were increased by 31.2% and 12.3% in acute study, whereas were decreased by 27.5% and 23.0% in chronic study. An attempt was made to determine if there were any influences on enzyme activities in rats pretreated with Capsicum extract chronically. Microsomal fraction was isolated from rat liver and quantity of cytochrome $P_{450}$ and $b_5$ in the microsomal fraction were determined by Omura's method. It was found that the quantity of cytochome $P_{450}$ was increased by 22.4%. The results suggest that microsomal drug metabolizing enzyme may be induced by chronic administration of Capsicum component, whereas it may be inhibited by acute administration of Capsicum component.

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Plant Inositol Signaling - Biochemical Study of Phospholipase C and D-myo-inositol -1,4,5-trisphosphate receptor

  • Martinec, Jan;Feltl, Tomas;Nokhrina, Katerina;Zazimalova, Eva;Machackova, Ivana
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.5
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    • pp.375-377
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    • 2000
  • It is now generally accepted that a phosphoinositide cycle is involved in the transduction of a variety of signals in plant cells. In animal cells, the hydrolysis of phosphatidyl-4,5-bisphosphate catalysed by phosphatidylinositol - specific phospholipase C yields to D-myo-inositol - 1,4,5-trisphosphate and diacylglycerol, which are well known second messengers. The binding of InsP$_3$to a receptor located on the endoplasmic reticulum triggers a calcium release from the endoplasmic reticulum. We have detected and partially characterised key components of phosphoinositide signaling. First, tobacco microsomal fraction and plasma membrane PI-PLC. Consecutively, using a radioligand binding assay we have identified a $Ca^{2+}$ -dependent high affinity InsP$_3$binding site in microsomal membrane fraction vesicle preparation and then we have measured inositol-1,4,5-trisphosphate induced calcium release from tobacco microsomal fraction. These findings suggest that phosphoinositide signaling system is present and operates in the tobacco suspension culture.e.

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The influence of Calcium, Magnesium, Sodium and Potassium Ion on the Adenosintriphosphatase activity of heart and skeletal microsomal fraction of rabbits. (가토심장(家兎心臟) 및 골격근(骨格筋)에서 분리(分離)한 Microsome 분획내(分劃內) ATPase 활성도(活性度)에 대(對)한 $Mg^{++}$, $Ca^{++}$, $Na^+$$K^+$의 영향(影響))

  • Choi, Sin-Jyoung;Hong, Ki-Whan;Kim, Gui-Tae
    • The Korean Journal of Pharmacology
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    • v.2 no.1 s.2
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    • pp.31-40
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    • 1966
  • The microsomal fraction is isolated from rabbit heart and skeletal muscle. The fraction is found to contain the $Na^+$-and $K^+$-activated ATPase. The maximal ATPase activity is obtained in $Na^+$ and $K^+$ concentration of 100 mM. Calcium itself stimulates the $Na^+$-and $K^+$-activated portion of ATPase in the presence of $Mg^{++}$. However, calcium does not stimulate ATPase in the absence of $Mg^{++}$.

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