• Title/Summary/Keyword: Marker enzyme

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Screening for Korean Vegetables with Anticarcinogenic Enzyme Inducing Activity Using Cell Culture System

  • Kim, Su-Mi;Ryu, Seung-Hee;Park, Hui-Don;Kim, Sung-Su;Kim, Jeong-Hwan;Kim, Jong-Sang
    • Preventive Nutrition and Food Science
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    • v.3 no.3
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    • pp.277-281
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    • 1998
  • There is extensive evidence suggesting the protective role of fruits and vegetables against chemically induced carcinogenesis. We have tested the ability of a representative range of Korean vegetables to act as blocking agents against neoplastic initiation by determining the induction level of quinone reductase , an anticarcinogenci marker enzyme, in hepalclc 7 cells exposed to vegetable extracts. Among thirty vegetables tested, Arcitum lappa(Burdock), Brassica juncea (Mustard leaf), Pteridium aguilinum (Bracken) and Chrysanthemum cornoratium(Crown daisy) caused a significant induction of quinone teductase activity with a limited increase in arylhdrocarbon hydroxylase activity. Combination of crown daisy with burdock had synergistic effect on quinone reductase induction. Quinone reductase-inducing activity was found mostly in hesane and ehtylactate fractions of MeOH extract of crown daisy while it ws not quinone reductase activity in liver, kideny, lung, and small intestine, confirming the presence of potent QR inducer (s) in crown daisy. These sata suggest that some vegetables including crown daisy induced QR merits further investigation as a potential cancer preventive agent in human.

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Total Cholesterol and Alkaline Phosphatase are Increased in D/D Type of Angiotensin Converting Enzyme

  • Kim, Sung-Su
    • Biomedical Science Letters
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    • v.18 no.4
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    • pp.406-412
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    • 2012
  • The polymorphism (insertion, I or deletion, D) of angiotensin converting enzyme (ACE) gene is designated as the presence of a 287 bp Alu repeat. The D/D homozygote carrier is associated with high ACE activity, and this high activity has been implicated with hypertension, coronary artery disease, or diabetic nephropathy. We studied the clinical candidate marker in ACE gene polymorphism using chemical and hematological analysis. The subjects are divided into normotensive and hypertensive groups and ACE genotype in the group was confirmed by PCR method. Chemical analysis was preceded with Hitachi7060, and hematological analysis was performed using Mythic 22. In 116 targeted people, 17 (38.64%) of 44 I/I genotype group are hypertension, 15 (34.09%) in 44 with D/I, but, D/D type in the 28 cases is 15 patients (53.57%) in hypertension. In hypertension group, biochemical analysis (triglyceride, and alkaline phosphatase) and hematological analysis (white blood cell, platelet) are showed high value in D/D genotype of ACE gene. The relationship between hypertension and ACE genotype is the same results as previously reported and we thought that the high laboratory value of white blood cell, platelet, triglycerides, and alkaline phosphatase are also indicator of hypertension in D/D type of ACE.

Application of Polyaniline to an Enzyme-Amplified Electrochemical Immunosensor as an Electroactive Report Molecule

  • Kwon, Seong-Jung;Seo, Myung-Eun;Yang, Hae-Sik;Kim, Sang-Youl;Kwak, Ju-Hyoun
    • Bulletin of the Korean Chemical Society
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    • v.31 no.11
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    • pp.3103-3108
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    • 2010
  • Conducting polymers (CPs) are widely used as matrixes for the entrapment of enzymes in analytical chemistry and biosensing devices. However, enzyme-catalyzed polymerization of CPs is rarely used for immunosensing due to the difficulties involved in the quantitative analysis of colloidal CPs in solution phase. In this study, an enzyme-amplified electrocatalytic immunosensor employing a CP as a redox marker has been developed. A polyanionic polymer matrix, $\alpha$-amino-$\omega$-thiol terminated poly(acrylic acid), was employed for precipitation of CP. The acrylic acid group acts as a polyanionic template. The thiol terminus of the polymer was used to produce self-assembled monolayers (SAMs) on Au electrodes and the amine terminus was employed for immobilization of biomolecules. In an enzymeamplified sandwich type immunosensor, the polyaniline (PANI) produced enzymatically is attracted by the electrostatic force of the matrix polymer. The precipitated PANI was characterized by electrochemical methods.

A Gene-based dCAPS Marker for Selecting old-gold-crimson (ogc) Fruit Color Mutation in Tomato (토마토 과색 돌연변이 유전자(old-gold-crimson) 선발을 위한 dCAPS 분자표지 개발)

  • Park, Young-Hoon;Lee, Yong-Jae;Kang, Jum-Soon;Choi, Young-Whan;Son, Beung-Gu
    • Journal of Life Science
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    • v.19 no.1
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    • pp.152-155
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    • 2009
  • The old-gold-crimson ($og^c$) fruit color mutation produces deep red tomato fruit with high lycopene content. age is a null mutation allele of lycopene-${\beta}$-cyclase (Crt-b) gene (B locus) that converts lycopene to ${\beta}$-carotene in the cartenoid biosynthesis pathway in tomato. Breeding of high lycopene tomato cultivars can be accelerated by marker-assisted selection (MAS) for introgression of $og^c$ allele by using a gene-based DNA marker. In order to develop a marker, single nucleotide deletion of adenine(A) with. in a poly-A repeat that has been known to be responsible for frame-shift mutation of $og^c$ was confirmed by resequencing mutant allele and wild-type allele at B locus of several tomato lines. For allele discrimination and detection of $og^c$, derived CAPS (dCAPS) approach was used by designing a primer that artificially introduced restriction enzyme recognition site of Hin fI in PCR products from $og^c$ allele. This dCAPS marker is co-dominant gene-based PCR marker that can be efficiently used for MAS breeding program aiming the development of high lycopene tomato.

Identification of a Causal Pathogen of Watermelon Powdery Mildew in Korea and Development of a Genetic Linkage Marker for Resistance in Watermelon (Citrullus lanatus)

  • Han, Bal-Kum;Rhee, Sun-Ju;Jang, Yoon Jeong;Sim, Tae Yong;Kim, Yong-Jae;Park, Tae-Sung;Lee, Gung Pyo
    • Horticultural Science & Technology
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    • v.34 no.6
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    • pp.912-923
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    • 2016
  • Watermelon production is often limited by powdery mildew in areas with a large daily temperature range. Development of resistant watermelon cultivars can protect against powdery mildew; however, little is known about the characteristics of its causal agents. Here, we identified the genus and race of a causal pathogen of powdery mildew in Ansung province of South Korea, and developed molecular markers for the generation of resistant watermelon cultivars. The causal pathogen was determined to be Podosphaera xanthii based on multiple sequence alignments of internal transcribed spacers (ITS) of rDNA. The physiological race was identified as 1W, and the Ansung isolate was named P. xanthii 1W-AN. Following inoculation with the identified P. xanthii 1W-AN, we found inheritance of the resistant gene fitting a single dominant Mendelian model in a segregated population ('SBA' ${\times}$ PI 254744). To develop molecular markers linked to fungus-resistant loci, random amplified polymorphic DNA (RAPD) was accomplished between DNA pooled from eight near-isogenic lines (NILs; $BC_4F_6$), originated from PI 254744 and susceptible 'SBB' watermelon. After sequencing bands from RAPD were identified in all eight NILs and PI254744, 42 sequence-characterized amplifiedregion (SCAR) markers were developed. Overall, 107 $F_2$ plants derived from $BC_4F_6$ NIL-1 ${\times}$ 'SBB' were tested, and one SCAR marker was selected. Sequence comparison between the SCAR marker and the reference watermelon genome identified three Nco I restriction enzyme sites harboring a single nucleotide polymorphism, and codominant cleavage-amplified polymorphic site markers were subsequently developed. A CAPS marker was converted to a high-resolution melt (HRM) marker, which can discriminate C/T SNP (254PMR-HRM3). The 254PMR-HRM3 marker was evaluated in 138 $F_{2:3}$ plants of a segregating population ('SBA' ${\times}$ PI254744) and was presumed to be 4.3 cM from the resistance locus. These results could ensure P. xanthii 1W-AN resistance in watermelon germplasm and aid watermelon cultivar development in marker-assist breeding programs.

Effects of Genistein Supplementation on Fatty Liver and Lipid Metabolism in Rats Fed High Fat Diet (고지방식이를 섭취하는 흰 쥐에서 제니스테인 보충이 지방간 및 지질대사에 미치는 영향)

  • Lee, Seon-Hye;Kim, Mi-Hyun;Park, Mi-Na;Lee, Yeon-Sook
    • Journal of Nutrition and Health
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    • v.40 no.8
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    • pp.693-700
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    • 2007
  • This study was performed to investigate the effects of genistein, a kind of soy isoflavones, on fatty liver and lipid metabolism in rats fed high fat diet. Twenty four male Sprague-Dawley rats were divided into four groups by dietary fat and genistein contents then raised for six weeks. The rats(n=6/group) were fed normal fat diet(NOR), high fat diet (HF), high fat with 0.1% genistein(HF+0.1%G) or high fat with 0.2% genistein(HF+0.2%G). Hepatic total lipid, triglyceride, total cholesterol and Serum GPT, as a marker for fatty liver, were significantly increased by high fat diet. Also, serum total lipid, triglyceride, total cholesterol, glucose and insulin concentration, hepatic lipogenic enzyme (fatty acid synthase and malic enzyme) activities were significantly increased by high fat diet. However, hepatic total lipid, triglyceride, total cholesterol and Serum GPT were significantly decreased by genistein intake. Also, genistein supplementation decreased serum total lipid, triglyceride, glucose and insulin concentration, hepatic lipogenic enzyme (fatty acid synthase and malic enzyme) activities. There were no differences by genistein level except for serum insulin. These results suggest that fatty liver induced by high fat diet was caused by increased serum lipid profiles and hepatic lipogenesis, whereas, genistein may be useful in inhibiting of fatty liver by reducing serum lipid profiles and hepatic lipogenesis.

Visible and Fast Assay System for Tobacco Transformant Introduced with Adenosine Deaminase Marker Gene (Adenosine Deaminase 표지유전자로 형질전환된 연초의 신속한 Assay 방법)

  • 양덕춘;김용환;임학태;방극수;배창휴
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.165-171
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    • 2001
  • New visible and fast assay system have been developed for tobacco transformant introduced with adenosine deaminase (ADA) marker gene, which converts cytotoxic adenosine analogues to non-toxic inosine analogues and ammonia. Ammonia was changed to blue color in the solution of phenol-nitoprusside and alkaline-hypochlorite. It was possible to detect activity of ADA visibly on the holes of 96 well plate using tiny explant of transgenic tobacco leaves within 1 hour incubation time. As substrates of ADA enzyme from transgenic plant on the plate, a number of adenosine analogues such as 9-D-arabinofuranosyl adenine, cordycepin, 2'-deoxyadenosine, adenosine and xylofuranosyl adenine were possible for detection of ADA activity. Optimal condition of substrate for ADA enzyme was each 10 mM and pH 7.5 in adenosine solution. Especially, transgenic plant did not convert adenosine to inosine and ammonia in the presence of ADA inhibitor deoxycoformycin, which means that ammonia produced from transgenic plant is due to expression of ADA gene. Now, we show that this detection system can be easily, sensitively, fast and cheaply as well as visibly assayed in vitro as GUS gene system with very small size of transformant explant.

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Induction of Quinone Reductase, an Anticarcinogenic Marker Enzyme, by Extract from Chrysanthemum zawadskii var. latilobum K.

  • Kim, Ju-Ryoung;Kim, Jung-Hyun;Lim, Hyun-Ae;Jang, Chan-Ho;Kim, Jang-Hoon;Kwon, Chong-Suk;Kim, Young-kyun;Kim, Jong-Sang
    • Preventive Nutrition and Food Science
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    • v.10 no.4
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    • pp.340-343
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    • 2005
  • Induction of NAD(P)H:(quinone-acceptor) oxidoreductase (QR) which promotes obligatory two electron reduction of quinones and prevents their participation in oxidative cycling and thereby the depletion of intracellular glutathione, has been used as a marker for chemopreventive agents. Induction of phase II enzyme is considered to be an important mechanism of cancer prevention. In our previous study, we assessed the quinone reductase QR-inducing activities of 216 kinds of medicinal herb extracts in cultured murine hepatoma cells, BPRc1 and hepalc1c7 cells. Among the 216 herbal extracts tested in that study, extracts from Chrysanthemum zawadskii showed significant induction of QR. In this study, we examined QR-inducing activity of solvent fractions of the herbal extract. The dichloromethane fraction of the herb showed the highest QR induction among the samples fractionated with four kinds of solvents with different polarity. The fraction also significantly induced the activity of glutathione S-transferase (GST), one of the major detoxifying enzymes, at $4{\mu}g/mL\;and\;2{\mu}g/mL$ in hepalc1c7 and BPRc1 cells, respectively. In conclusion, dichloromethane-soluble fraction of Chrysanthemum zawadskii which showed relatively strong induction of detoxifying enzymes merits further study to identify active components and evaluate their potential as cancer preventive agents.

Effects of Commelina communis L. on the Blood Glucose Level in Alloxan Induced Diabetic Rat and the Biochemical Properties of Glucose-6-Phosphate Dehydrogenase from the Rat Livers (당뇨유발쥐에서 닭의장풀의 혈당감소효과와 간조직내의 Glucose-6-Phosphate Dehydrogenase의 효소활성에 미치는 효과)

  • Park, Soo-Young;Cho, Kyung-Hea
    • Korean Journal of Pharmacognosy
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    • v.25 no.3
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    • pp.238-248
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    • 1994
  • The hypoglycemic and metabolic effects of Commelina communis L. extract were investigated in alloxan induced diabetic rats. The increased blood glucose level in the diabetic rats was significantly reduced and the loss of body weight was recovered with the treatment of the plant protein fractions($30{\sim}70%$ ammonium sulfate precipitates). Administration of the plant protein fractions elicited the significant increase of glucose-6-phosphate dehydrogenase (G-6-P DH) activity and liver weight which were decreased in the diabetic rat liver. G-6-P DH was partially purified from extract- or insulin-treated diabetics, diabetic control, and normal rat liver and studied for the biochemical properties. The $K_m$ value(9.002 mM) of diabetic rat liver enzyme was greatly higher than that (0.033 mM) of normal enzyme indicating the affinity of enzyme for the substrate was significantly reduced in the diabetic rat liver. This reduced affinity of enzyme for the substrate in the diabetic rat was recovered in the extract- or insulin-treated rat liver enzyme having 0.164 or 0.208 mM of their $K_m$ values, respectively. Although there was no significant difference in the optimum pH(6.0) and optimum temperature($37^{\circ}C$) of enzyme among the experimental groups, the dependence of their activities on pH appeared to be slightly resistant in the extract- or insulin-treated group compared to the diabetic group. In order to investigate the antigenicity of rat liver enzyme among experimental groups, enzyme-linked immunosorbent assay was carried out by using anti-G-6-P DH anti-serum. Absorbance(0.102) shown in the normal rat liver was reduced even below zero in the alloxan-diabetic rat liver, but increased again in the extract- or insulin-treated rat liver(0.096 or 0.118, respectively). The result of this study suggested that G-6-P DH may be used as a marker enzyme to diagnose and to indicate the progress of the diabetics, and the hypoglycemic effect of the extracts of Commelina communis L. was certainly associated with action or mode of G-6-P DH on the rat liver.

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Cardioprotective effect of Argyreia speciosa (Burm. f) Boj. extracts against Isoproterenol- induced myocardial infarction in rats

  • Thakker, Shalin;Biradar, S.M.;Habbu, P.V.;Mahadevan, K.M.;Thippeswamy, B.S.;Veerapur, V.P.
    • Advances in Traditional Medicine
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    • v.10 no.4
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    • pp.278-287
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    • 2010
  • The present study was undertaken to evaluate the protective effect of ethanol (EtAS), ethyl acetate (EAAS) and aqueous (AQAS) extracts of Argyreia speciosa (AS) roots against Isoproterenol (ISO)-induced myocardial infarction in rats. The animals were exposed to isoproterenol (200 mg/kg. s.c) twice at an interval of 24 hrs. Cardioprotective effect was assessed by observing ECG parameters, serum marker enzymes and histopathology of the heart. Pretreatment of EAAS, and EtAS (200 mg/kg) resulted in a significant (P < 0.001) increase in P wave, QRS complex and R-R interval, whereas heart rate, QT interval and cardiac cycle were maintained near to normal values. EtAS and EAAS showed significant (P < 0.05; P < 0.001) reduction in all the tested diagnostic markers compared to ISO treated group. Histological studies on the structural changes of heart tissue supported the protective activity of AS. The result suggest that treatment of AS prior to ISO has a significant role in protecting the animals from ISO induced myocardial infarction.