• Title/Summary/Keyword: isozymes

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Reduction of Bacterial Mutagenesis of 2-Amino-3-Methylimidazo[4,5-f]quinoline by S-9 Fraction from Mice Treated with Conjugated Linoleic Acid (CLA)

  • Park, Kyung-Ah;Kim, Seck-Jong;Park, Soo-Jahr;Park, Gu-Boo;Lim, Dong-Kil;Bahn, Kyeong-Nyeo;Cho, Yong-Un;Park, Jung H.Y.;Pariza, Michael W.;Ha, Yeongl-Lae
    • Preventive Nutrition and Food Science
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    • v.6 no.1
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    • pp.57-61
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    • 2001
  • Conjugated linoleic acid (CLA), when incorporated into mouse liver microsomal membranes, selectively inhibits the mutagenesis of 2-amino-3-methylimidazo[4,5-f] quinoline (IQ). Nine-week old female ICR mice were given (p.o.) 0.1 mL olive oil alone (control), 0.1 mL olive oil plus 0.1 mL linoleic acid, or 0.1 mL olive oil plus 0.1 mL CLA, twice weekly for four weeks. The animals were then sacrificed and liver S-9 fractions were prepared. Activation of IQ for mutagenesis by the liver S-9 from CLA-treated mice was significantly reduced in comparison wit liver S-9 from control or linolic acid-treated mice. By contrast, the activation of 7,12-dimethylbenz[a] anthracene (DMBA) and benzo[a] pyrene (BP) was unaffected. Hence, CLA incorporated into phospholipids may selectively affect cytochrome P450 isozymes responsible for activating IQ, but not those which activate BP or DMBA. The addition of free CLA or the methyl esters of CLA, linoleic acid, or oleic acid, to control S-9 inhibited the activation of all three mutagens (IQ, BP, and DMBA).

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Scavenging Activity of Reactive Oxygen Species and Inhibitory Effect of Cytochrome P450 from Circium japonicum Extract (대계 추출물의 할성산소 소거능 및 Cytochrome P450 효소 저해효과)

  • Kim, Hyuck;Yi, Hyo-Seung;Park, Won-Hwan;Moon, Jin-Young
    • The Korea Journal of Herbology
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    • v.22 no.1
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    • pp.53-61
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    • 2007
  • Objectives: Our previous studies have clearly demonstrated that the scavenging activity of reactive oxygen species (ROS), protective effect of lipid peroxidation (LPO), and inhibition of cytochrome P450 isozymes (CYPs) from the Circium japonicum aqua-acupuncture solution (CJAS). But, Circium japonicum water extracted solution (CJWS) was weakly reported in cardiovascular diseases such as oxidative stress-mediated atherosclerosis or its value evaluated. Methods: CJWS was assessed to determine the mechanism of its scavenging activity of ROS and inhibitory effect of CYP 2E1. Results: CJWS exhibited a concentration-dependent scavenger of DPPH and superoxide anions radicals using different assay systems. In addition, CJWS showed dose-dependent free radical scavenging activity, including hydroxyl radicals, peroxynitrite, and nitric oxide. The CJWS was also found to be effective in protecting rat liver homogenate against LPO. Futhermore, the CJWS showed significant inhibition of CYP 2E1 induced by pyrazol in a rat liver microsome. Conclusion : ROS and CYPs may play a role in several diseases, such as cardiovascular disease and heart failure. Our study demonstrated that the CJWS has excellent scavenging activity of ROS. Hence, it is worthwhile to investigate the potential effectiveness of CJWS in preventing oxidative stress-mediated cardiovascular diseases.

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Genetic Characterization of Two Types of Sea Bass, Lateolabrax japonicus in Korea by Isozyme Analysis (Isozyme 분석에 의한 한국산 농어, Lateolabrax japonicus 2형간의 유전학적 특징)

  • Park Jung-Youn;Kim Kyung-Kil;Kim Yoon
    • Journal of Aquaculture
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    • v.9 no.4
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    • pp.437-444
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    • 1996
  • Genetic characterization and identification of two types of sea bass Lateolabrax japonicus were performed by examining electrophoretic patterns of isozymes. Twenty five loci coding for thirteen enzymes were detected in two types. Among the twenty five loci, one completely divergent loci (Pt-1) was observed between two types. Nei's genetic distance between two types was 0.12036. The estimated divergence time of these two types may have about $6.2{\times}10^5$ years ago. On the other hands, the expected average heterozygosity was 0.084 in not spotted type on the body surface and 0.067 in spotted type on the body surface. These results mean that the existance of two types of sea bass was established in present study which may have had genetic divergence.

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Inhibitory Properties of Nerve-Specific Human Glutamate Dehydrogenase Isozyme by Chloroquine

  • Choi, Myung-Min;Kim, Eun-A;Choi, Soo-Young;Kim, Tae-Ue;Cho, Sung-Woo;Yang, Seung-Ju
    • BMB Reports
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    • v.40 no.6
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    • pp.1077-1082
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    • 2007
  • Human glutamate dehydrogenase exists in hGDH1 (housekeeping isozyme) and in hGDH2 (nerve-specific isozyme), which differ markedly in their allosteric regulation. In the nervous system, GDH is enriched in astrocytes and is important for recycling glutamate, a major excitatory neurotransmitter during neurotransmission. Chloroquine has been known to be a potent inhibitor of house-keeping GDH1 in permeabilized liver and kidneycortex of rabbit. However, the effects of chloroquine on nerve-specific GDH2 have not been reported yet. In the present study, we have investigated the effects of chloroquine on hGDH2 at various conditions and showed that chloroquine could inhibit the activity of hGDH2 at dose-dependent manner. Studies of the chloroquine inhibition on enzyme activity revealed that hGDH2 was relatively less sensitive to chloroquine inhibition than house-keeping hGDH1. Incubation of hGDH2 was uncompetitive with respect of NADH and non-competitive with respect of 2-oxoglutarate. The inhibitory effect of chloroquine on hGDH2 was abolished, although in part, by the presence of ADP and L-leucine, whereas GTP did not change the sensitivity to chloroquine inhibition. Our results show a possibility that chloroquine may be used in regulating GDH activity and subsequently glutamate concentration in the central nervous system.

EFFECTS OF NOVEL DITHIOL MALONATE DERIVATIVES ON LIVER LIPID PEROXIDATION AND ON MICROSOMAL ELECTRON TRANSPORT SYSTEM

  • Park, Keun-Hee;Lee, Jong-Wook
    • Toxicological Research
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    • v.3 no.2
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    • pp.97-110
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    • 1987
  • The effects of 5 novel hepatotrophic agents, dithiol malonate derivatives (DMDs; DMD1-DMD5), on the liver microsomal lipid peroxidation induced by carbon tetrachloride $(CCl_4)$ and the correlations with the changes of microsomal electron transport system were investigated. All DMDs were found to inhibit the lipid peroxidation induced by $CCl_4$ in mice and rats as well in vitro liver microsomal system. Therefore, each DMD seemed to have direct mode of action on liver microsomes to inhibit the lipid peroxidation. As an ex vivo study, the induced lipid peroxidation by $CCl_4$ and the changes in electron transport system were determined with liver microsomes obtained from rats chronically treated with DMDs for 7 days. The induced lipid peroxide contents in liver microsomal system were lower in DMD1, DMD2 and DMD3 treated group, but higher in DMD4 and DMD5 group when compared to the control group. Cyt. p.450 contents in the microsomes were decreased by the treatment with DMD1, DMD2 and DMD3, but increased significantly by DMD4 with great extent and by DMD5 with less extent. The cyt. p-450 isozymes induced by treatment of DMD4 and DMD5 were identified as 3-methylcholanthrene (MC) type. The NADPH cyt. -C reductase activities of the microsomes treated with DMD1, DMD2, DMD4 and DMD5 were increased in the range of around 20% to 50%, but decreased with DMD3, All DMDs increased dyt. $-b_5$ content and did not alter NAdH-cyt, $-b_5$ reductase activities in the microsomes. In summary, the 5 novel hepatotrophic agents (DMDs) markedly protected against lipid peroxidation induced by $CCl_4$ in vivo and in vitro possibly through the mechanism of direct action on the liver microsomes. The degree of inhibition produced by DMDs on lipid peroxidation induced by $CCl_4$ seemed to coincide rather with cyt. p-450 contents than with other components of liver microsomal electron transport system including NADPH-cyt, -C reductase.

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Suppression of Lipid Peroxidation and CYP Isozymes activities by Circium japonicum Herbal-acupuncture Solution ; Basic Study for Screening of Medicinal Herb on Reactive Oxygen Radical and CYP-Mediated Atherosclerosis (대계 약침액(藥鍼液)의 지질과산화 및 CYP 억제에 미치는 영향(影響) ; 활성산소자유기 및 CYP 매개의 동맥경화 치료를 위한 천연약물 개발의 기초 평가)

  • Lee Jeong-Joo;Kim Hyuck;Yi Hyo-Seung;Park Won-Hwan;Moon Jin-Young
    • Korean Journal of Acupuncture
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    • v.23 no.4
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    • pp.177-186
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    • 2006
  • 목적 : 약침액(藥鐵液)의 지질과산화 예방 및 cytocome P450과의 상호 작용에 있어서 대계의 역할은 과거 연구가 거의 없었다. 따라서 본 실험에서는 대계 약침액이 지질과산화를 예방하고, 심혈관계질환 유발에 밀접한 연관이 있는 cytochrome P450의 직접적인 저해 효과를 검토 하고자 한다. 방법 : 대계 약침액이 지질과산화를 억제하는 정도를 평가하기 위하여 세포막을 구성하는 불포화지방산의 일종인 linoleic acid를 대상으로 지질과산화 진행 시간과 대계 약침액의 농도에 의존적인 저해 효과를 실험하였다. 또한 실험쥐의 간조직을 이용하여, 강제적인 과산화를 유도한 후 이를 방어하는 효능을 검토하였다. 그리고 cytochrome P450을 구성하는 그룹의 1A1, 1A2 및 2E1의 활성을 각각 EROD, MROD, p-nitrophenol, aniline 방법으로 측정하였다. 결과 및 결론 : 대계 약침액은 세포막 구성의 불포화 지방산인 linoleic acid의 산화를 시간 및 처리 농도에 의존적으로 억제하였고, 실험쥐의 조직 과산화를 유의성 있게 저해하였다. 또한 aryl hydrocarbon receptor (AHR)을 활성화 시켜 polycyclic aromatic hydrocarbons (PAHs)에 의한 심혈관계 질환 유발 인자로 알려진 cytochrome P450 1A1 및 1A2의 발현을 일부 저해하였으며, 특히 체내에 흡수된 알콜 대사에 관여하는 P450 2E1을 강하게 억제 시켰다.

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No Role of Protected Region B of Human Cytochrome P4501A2 Gene (CYP1A2) As an AP-1 Response Element

  • Chung, In-Jae;Jung, Ki-Hwa
    • Archives of Pharmacal Research
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    • v.25 no.3
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    • pp.375-380
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    • 2002
  • Cytochrome P4501A2 (CYP1A2) is a member of the cytochrome P450 family of isozymes involved in the phase I drug metabolism of vertebrates. CYP1A2 is responsible for the activation of a number of aromatic amines to mutagenic and carcinogenic forms. Thus, the level of CYP1A2, which varies among different populations, may determine an individual's susceptibility to these chemicals. We have previously reported on the importance of a cis element named PRB (protected region B) in the regulation of human Cytochrome P4501A2 (CYP1A2) gene, which appeared to act as a positive regulatory element. Closer examination of the PRB sequence (-2218 to -2187 bp) revealed a putative AP-1 binding site, TGACTAA, at -2212 bp (Chung and Bresnick, 1997). To elucidate the role of AP-1 in CYP1A2 regulation, we transiently overexpressed c-Jun and c-Fos transcription factors in human hepatoma HepG2 cells, and examined their influence on the CYP1A2 promoter activity by reporter gene assays. Cotransfection of the c-Jun and the c-Fos expression vectors increased the induced transactivation by five to six fold from the CYP1A2 promoter constructs. However, deletion of the PRB element did not affect the degree of activation by the c-Jun and the c-Fos. Therefore, it is unlikely that the c-Jun and the c-Fos activate the CYP1A2 promoter through this AP-1 consensus-like sequence in the PRB region.

Isozymic Characteristics of Multiple-Ear and Tiller Maize Lines (다수다얼성 옥수수의 동위효소 특성)

  • ;Bong-Ho Chae
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.32 no.1
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    • pp.16-23
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    • 1987
  • This experiment was conducted to determine the isozymic differences between normal maize and maize inbreds of multiple ears and tillers (MET). Two maize inbreds Euisung, Iri and their hybrid having tillers and multiple ears were compared with normal maize. With usual electrophoresis using 6% polyacrylamide gel, peroxidase and esterase enzymes were studied. Matured leaf, culm, leaf sheath, root and young ear tissues showed different isozymic patterns between METs and normal maize in peroxidase. The Euisung inbred grown for 7 days under dark condition showed typical peroxidase. bands compared with checks in the tissues of coleoptile and stele. Better observation of isozymic bands was made during early part of maize growth. Parental inbreds showed more active and apparent band differences than their hybrids in esterase. Bands for esterase were also apparently different in the stele, coleoptile and young ear tissues of the METs and the checks. The maize lines infected with black streaked dwarf virus showed obvious differences in peroxidase and esterase isozymes.

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Different oxidative burst patterns occur during host and nonhost resistance responses triggered by Xanthomonas campestris in pepper

  • Kwak, Youn-Sig;Han, Ki-Soo;Lee, Jung-Han;Lee, Kyung-Hee;Chung, Woo-Sik;Mysore, Kirankumar S.;Kwon, Young-Sang;Kim, Hee-Kyu;Bae, Dong-Won
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.244-254
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    • 2009
  • The hypersensitive reaction (HR) is the most common plant defense reaction against pathogens. HR is produced during both host- and nonhost-incompatible interactions. Several reports suggest that similarities exist between host and nonhost resistances. We assayed the pattern of generation of reactive oxygen species (ROS) and scavenging enzyme activities during nonhost pathogen-plant interactions (Xanthomonas campestris pv. campestris/Capsicum annuum L.) and incompatible host pathogen-plant interactions (Xanthomonas campestris pv. vesicatoria race1/Capsicum annuum L.). Both ${O_2}^-\;and\;H_2O_2 $ accumulated much faster during nonhost resistance when compared to the host resistance. The scavenging enzyme activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX) were also different during the host- and nonhost-incompatible interactions. CAT activity was much higher during nonhost resistance, and several new isozymes of SOD and POX were detected during nonhost resistance when compared to the host resistance. Lipoxygenase (LOX) activity was higher in host resistance than nonhost resistance during the early stages of infection. Interestingly, the nitric oxide (NO) radical accumulated equal amounts during both host and nonhost resistance at early stages of infection. Further studies are needed to determine the specific pathways underlying these differences between host and nonhost resistance responses.

Toxicological Aspects of Carboxylesterases -A Sensitive Biomarker of Organophosphate Toxicity-

  • Satoh, Tetsuo;Suzuki, Satoshi;Hosokawa, Masakiyo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.108-113
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    • 1998
  • Egasyn is accessory protein of ${\beta}$-glucuronidase(${\beta}$-G) in the liver microsomes. Liver microsomal ${\beta}$-G is stabilized within the luminal site of the microsomal vesicles by complexation with egasyn which is one of carboxylesterase isozymes. We investigated the effects of organophosphorus compounds(OPs) such as insecticides on the dissociation of egasyn-${\beta}$-glucuronidase(EG) complex. The EG complex was easily dissociated by administration of OPs, i.e., Fenitrothion, EPN, Phenthionate, and bis-p-nitrophenyl phosphate(BNPP), and resulting ${\beta}$-G dissociated was released into blood, leading to the rapid and transient increase of plasma ${\beta}$-G level with a concomitant decrease of liver microsomal ${\beta}$-G level. In a case of phenthionate treatment, less increase in plasma ${\beta}$-G level was observed, as compared with those of other OPs. This may be explained by a fact that phenthionate was easily hydrolyzed by carboxylesterase. Similarly, carbamate insecticides such as Carbaryl caused rapid increase of plasma ${\beta}$-G level. In contrast, no significant increase of plasma ${\beta}$-G level was observed when pyrethroid insecticides were administered to rats. This is due to a fact that pyrethroids such as Phenthrin and Allethrin were easily hydrolyzed by A-esterase as well as carboxylesterase. On the other hand, addition of OPs to the incubation mixture containing liver microsomes caused the release of ${\beta}$-G from microsomes to the medium. From these in vivo and in vitro data, it is concluded that increase of the plasma ${\beta}$-G level after OPs administration is much more sensitive biomarker than cholinesterase inhibition to acute intoxication of OPs and carbamates.

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