• Title/Summary/Keyword: enzyme transport

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A study on the mechanism for reduction of lead-induced toxicity in nervous system by aloe vera (Aloe vera의 연 유도성 신경계 독성 저감 기전에 관한 연구)

  • 정명규;곽영규
    • Journal of Environmental Health Sciences
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    • v.22 no.3
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    • pp.8-16
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    • 1996
  • Effects of water extract of aloe vera on lead-induced neurotoxicity were investigated in sciatic nerve isolated from rat. The mechanism on toxicity reduction by measuring activities of axonal enzymes, metabolism of myo-inositol in nerve, lead concentration in several organs and so on were further examimed. In the lead-treated rats, the transport rate of axonal enzyme, such as acetyl cholinesterase and choline acetyltransferase, was reduced by from 50% to 30% respectively. Reduction in myo-inositol concentration and $Na^+/K^+$ ATPase activity were also observed in sciatic nerve from lead-treated rat. However, the aloe extract administration significantly eliminated the impairment and maintained myo-inositol concentration to about 85% of normal level. Also aloe extract promoted the excretion rate of lead which is accumulated in blood, sciatic nerve and kidney. These results suggest that lead-induced neurotoxicity was significantly reduced by administration of aloe extract and the mechanism might be partly increase in kidney excretion rate of lead and parity normalization of $Na^+/K^+$ ATPase activity which is critical factor in order to keep nerve maintaining normal myo-inositol level.

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Effects of Cadmium and Cadmium Binding Protein on the Respiration of Mitochondria of Rat Kidney (흰쥐 신장의 미토콘드리아 호흡에 미치는 Cadmium과 Cadmium Binding Protein의 영향)

  • Cho, Hae-Kyoung;Boo, Moon-Jong;Choe, Rim-Soon
    • Applied Microscopy
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    • v.19 no.2
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    • pp.59-73
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    • 1989
  • To investigate the effect of cadmium and cadmium binding protein on the electron transport system and conformational changes of rat kidney mitochondria, various cadmium concentration were treated in vitro and respiration rate, NADH-CoQ reductase activity were measured. Ultrastructural changes at state IV respiration were also observed. CdBP was isolated from the rat liver by Sephadex G-75 column fractionation and treated in vitro with cadmium. Also mitochondrial state IV respiration rate was measured. When cadmium was treated in vitro, state IV respiration and enzyme activity were decreased and ultrastructural transformation of mitochondria from a condensed to an orthodox conformation was inhibited under state IV respiration. In case cadmium and CdBP were treated together, oxygen consumption was more increased than cadmium only. Conformational changes of mitochondria from a condensed to orthodox conformation were also observed. This indicates that CdBP have a protective effect against cadmium toxicity.

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Studies on the Microbial Pigment (V) The effect of some detergent on pigment formation in Serratia marcescens strain P (微生物의 色素에 關한 硏究(第 5 報) -色素形成에 미치는 界面活性劑의 영향-)

  • Lee, Ho-Yong;Cho, Hong-Bum;Choi, Yong-Keel
    • Korean Journal of Microbiology
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    • v.22 no.3
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    • pp.191-195
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    • 1984
  • In order to study on the pigment formation of Serratia marcescens, the synthesis of prodigiosin was examined in the presence of a wide range of concentration of detergents. A high elevation of pigment formation was obtained in case of the treatment with SDC and SAP. And the population growth of the bacteria was increased by SDC and SAP, in the concentration of optimum concentration of pigment formation. The alkaline phosphatase activity was also increased in the treatment of SAP, SDC and SDS. The possible mechanism of the detergents on enhancement of pigment formation could be explained by an increase of enzyme activity and membrane transport.

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Synthesis of Lipophilic Benz[cd]indole Antifolates and Their Antitumor Activity In Vitro (지용성 Benz[cd]indole 엽산길항제의 합성 및 In Vitro 항암활성)

  • Baek Du-Jong
    • YAKHAK HOEJI
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    • v.49 no.1
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    • pp.60-67
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    • 2005
  • Nonclassical aminobenz[cd]indole antifolates 4, 5 and 6, in which the glutamic acid moiety of the classical antifolates is substituted by 2-phenylglycinamide or 3-aminobenzamide, were synthesized and their in vitro antitumor activity was evaluated. The purpose of this substitution is that the lipophilicity is enhanced due to the aromatic ring of the target compounds for the passive transport through lipid membrane of cells while the hydrogen bonding of the amide is retained in the active site of the enzyme, thymidylate synthase, where the glutamate is originally present. The target compounds were highly cytotoxic against tumor cell lines of murine and human origin with micromolar to nanomolar $IC_{50}$ values. Most effective was compound 4 ($N^6-methyl-N^6$-[4-[(${\alpha}$(S)-aminocarbonylbenzyl) aminocarbonyl]benzyl]-2,6-diaminobenz[cd]indole)with $IC_{50}$ of 2 nM against SW480, human colon adenocarcinoma cell line, which is 650-fold more potent than the reference compound 3.

Radioimmunoassay of High-Density Lipoprotein in Japanese Quail Serum (메추리혈청 리포단백질의 방사면역분석법)

  • 한병훈;한용남;유시용;양현옥;박명환
    • YAKHAK HOEJI
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    • v.29 no.2
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    • pp.90-95
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    • 1985
  • The development of radioimmunoassay (RIA) for high-density lipoprotein (HDL) in Japanese quail serum will contribute to the screening of drugs acting on cholesterol transport. We have developed a double antibody RIA method for J. quail HDL. The first antibody was raised in rabbit by immunization of HDL isolated by the dextrane sulfate-$Mn^{#}$ precipitation method. For the preparation of raclioiodinated antigen, HDL was further purified by combination of electrophoretic procedure. Using the second antibody raised in goat by rabbit IgG, we have furnished the RIA method for HDL. It showed high specificity and sensitivity of working assay range, 0.1-33.mu.g HDL/tube. There was no correlation between the radioimmunoassay of HDL and the enzyme assay of HDL-cholesterol.

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Microsomal Proton Transport Activity Measured by Quinacrine Fluorescence from Tomato Roots (Quinacrine 형광을 이용한 토마토 뿌리조직 마이크로솜의 수소이온이동 활성측정)

  • Shin, Dae-Seop;Cho, Kwang-Hyun;Kim, Young-Kee
    • Applied Biological Chemistry
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    • v.45 no.2
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    • pp.53-58
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    • 2002
  • Quinacrine, a pH-sensitive fluorescence probe, which exists either as an unprotonated fluorescence form or a protonated noufluorescence form, can be used to measure the proton transport activity of $H^+-ATPase$. Quinacrine was used to determine the optimal conditions for measuring the activity of microsomal $H^+-ATPase$ prepared from the roots of tomato plants. The amount of quinacrine fluorescence quenching obtained at $0.43{\mu}g/{\mu}l$ of microsomal protein concentration was 25-26%, which shows that the enzyme activity of 100 nmol/min decreases 10% of quinacrine fluorescence. Maximal fluorescence quenching was obtained at pH 7.0-7.2 and 2 mM $Mg^{2+}$ Because the activity of microsomal $H^+-ATPase$ is also maximal at these conditions, the quinacrine fluorescence well represents the activity of $H^+-ATPase$. Vanadate and $NO_3-$, specific inhibitors of plasma and vacuolar $H^+-ATPases$, respectively, were successfully applied to inhibit the quinacrine fluorescence quenching mediated by the corresponding $H^+-ATPases$. These results imply that quinacrine is a useful tool for measuring the proton transport activities of microsomes obtained from the root tissue of tomato plants.

Characteristics of p-Aminohippurate (PAH) Transport in Basolateral Membrane Vesicles of Rabbit Kidney Cortex -Effect of Anion- (신장 피질 기저막 소포에서 p-Aminohippurate 이동의 특성에 대한 연구 -음이온의 효과-)

  • Lee, Sang-Ho;Chung, Jin-Sup;Kim, Yong-Keun
    • The Korean Journal of Physiology
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    • v.20 no.2
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    • pp.225-235
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    • 1986
  • Effects of anions on p-Aminohippurate (PAH) transport across the basolateral membrane (BLM) were studied. Basolateral membrane vesicles were isolated from rabbit renal cortex by using a self-orienting Percoll-gradient centrifugation and $Mg^{2+}-precipitation$. The characteristics of the membrane vesicles was examined by marker enzyme activity, membrane orientation and transport studies. The Na-K-ATPase activity in the fraction containing BLM vesicles was enriched 9·fold, and the alkaline phosphatase activity in the fraction containing BBM vesicles was increased 9-fold, compared with those of the homogenate. The transport properties of the two membrane preparations were studied by a rapid filtration technique. The uptake of PAH by BLM was sensitive to changes in medium osmolarity and inhibited by probenecid. When the uptake of $50{\mu}M$ PAH in voltage-clamped BLM vesicles was determined in the presence of various anions in the incubation medium, cis inhibitions by $SO_4\;and\;SSO_3$ were observed in the presence of sodium gradient (out>in). Sodium-dependent PAH uptake was inhibited competitively by external $SO_4$ PAH uptake in BLM vesicles loaded with 20 mM acetate and $SO_4\;or\;200\;{\mu}M$ PAH was significantly stimulated as compared with unloaded vesicles. The extent of trans-stmulation of PAH uptake by $SO_4$. was increased with the inside concentration of $SO_4$. This trans-stimulatory effect by $SO_4$, was observed to be additive in the presence of Na gradient and completely inhibited by 2 mM probenecid and 1 mM SITS. These results demonstrate that PAH/anion exchange is present in BLM of renal cortex and in this exchange mechanism inorganic as well as organic anions are involved as substrates.

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Changes of Organic Solutes and Antioxidative Enzyme Activity in Rice Seedling under Salt Stress

  • Park So-Hyeon;Sung Jwa-Kyung;Lee Su-Yeon;Lee Ju-Young;Jang Byoung-Choon;Song Beom-Heom;Kim Tae-Wan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.5
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    • pp.325-331
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    • 2005
  • Seedlings of two rice genotyopes, cvs. Ilpumbyeo and Gancheokbyeo, were exposed to 0, 50 and 100 mM NaCl in nutrient solution for nine days. Plants were collected at the interval of 3 days and organic and inorganic solutes in leaves and roots and antioxidative enzyme activity in leaves were determined. Under salinity, the accumulation of soluble sugars occurred considerably in the older leaves of stressed seedlings compared to younger leaves and roots. The endogenous Na+ contents markedly increased at higher NaCl concentration in leaves and roots of seedlings, though it was higher accumulated in roots. Salinity resulted in an excessive proline accumulation in the stressed plants. A more pronounced increase was observed in Gancheokbyeo leaves. SOD activity in Impumbyeo cannot found any remarkable change, whereas, in Gancheokbyeo, its activity was rapidly decreased. CAT and POD activities increased with an increase in NaCl concentration in both genotypes. In sum­mary, the high capacity of rice seedlings to overcome an unfavorable growth condition such salt stress appears to be related to an adequate partition of organic solutes between shoots and roots and to changes in absorption, transport and re-translocation of salts.

Prenatal Diagnosis of Mucolipidosis Type II: Comparison of Biochemical and Molecular Analyses

  • Kosuga, Motomichi;Okada, Michiyo;Migita, Osuke;Tanaka, Toju;Sago, Haruhiko;Okuyama, Torayuki
    • Journal of mucopolysaccharidosis and rare diseases
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    • v.2 no.1
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    • pp.19-22
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    • 2016
  • Purpose: Mucolipidosis type II (ML II), also known as I-cell disease is an autosomal recessive inherited disorder of lysosomal enzyme transport caused by a deficiency of the uridine diphosphate (UDP)-N-acetylglucosamine:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase (GlcNAc-phosphotransferase). Clinical manifestations are skeletal abnormalities, mental retardation, cardiac disease, and respiratory complications. A severely and rapidity progressive clinical course leads to death before 10 years of age. Methods/Results: In this study we diagnosed three cases of prenatal ML II in two different at-risk families. We compared two procedures -biochemical analysis and molecular analysis - for the prenatal diagnosis of ML II. Both methods require an invasive procedure to obtain specimens for the diagnosis. Biochemical analysis requires obtaining cell cultures from amniotic fluid for more than two weeks, and would result in a late diagnosis at 19 to 22 weeks of gestation. Molecular genetic testing by direct sequence analysis is usually possible when mutations are confirmed in the proband. Molecular analysis has an advantage in that it can be performed during the first-trimester. Conclusion: Molecular diagnosis is a preferable method when a prompt decision is necessary.

Phenol Removal Using Horseradish Peroxidase(HRP)-Mediated Polymerization Reaction in Saturated Porous Media (다공성 포화 매질에서 효소 중합반응을 이용한 페놀 제거)

  • Kim, Won-Gee;Lee, Seung-Mok
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.10
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    • pp.984-991
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    • 2008
  • This paper reports experimental results, demonstrating the feasibility of horseradish peroxidase(HRP) and H$_2$O$_2$ to reduce phenol transport in saturated porous media. A laboratory-scale packed column reactor(ID: 4.1 cm, sand-bed height 12 cm) column was utilized to simulate injection of HRP and H$_2$O$_2$ into an aquifer contaminated with phenol. Effluent concentrations of phenol and polymerization products were monitored before and after enzyme addition under various experimental conditions(enzyme dose: 0$\sim$2 AU/mL, [ionic strength]: 5$\sim$100 mM, pH: 5$\sim$9). The concentration of phenol in the column effluent was found to decrease by nearly 90% in the presence of HRP(2 AU/mL) and H$_2$O$_2$ in the continuous flow system at pH 7 and ionic strength 20 mM. The influent phenol was converted in the system to insoluble precipitate, which deposited in pore spaces. The remains were discharged as soluble oligomers. About 8% of total pore volume in column system was decreased by deposition of polymer produced.