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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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Optimization of \beta-mammanase Production from Bacillus subtilis JS-1. (\beta-Mannanase를 생산하는 Bacillus subtilis JS-1의 분리 및 효소 생산성)

  • 임지수;정진우;이종수;강대경;강하근
    • Microbiology and Biotechnology Letters
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    • v.31 no.1
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    • pp.57-62
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    • 2003
  • A bacteria strain producing extracellular $\beta$-mannanase was isolated from soil and was identified as Bacillus subtilis by 16S rRNA sequence comparison and biochemical determinations. The optimum pH and temperature for the $\beta$-mannanase activity were 5.0 and 5.5$^{\circ}C$, respectively. The zymogram technique revealed a single protein band exhibiting $\beta$-mannanase activity from the culture supernatant. The molecular mass of the enzyme was estimated at approximately 130 kDa. The addition of 0.5% lactose or 0.5% locust bean gum to the LB medium caused to Increase significantly the $\beta$-mannanase productivity from Bacillus subtilis JS-1. The cells grown on LB medium supplemented with lactose produced maximal enzyme activity at the stationary phase. In contrast to this, the $\beta$-mannanase was induced at the logarithmic phase from the cells grown on LB medium supplemented with locust bean gum. The discrepancy in induction times suggests that $\beta$-mannanase was induced by different induction mechanisms depending on the carbon sources in Bacillus subtilis JS-1 .

Antitumor Effect of $18{\beta}$-Glycyrrhetinic Acid against Human Tumor Xenografts Caused by A549 Cancer Cell (A549 암세포 기인성 종양에 대한 $18{\beta}$-Glycyrrhetinic Acid의 항종양효과)

  • Kim, Ha-Yan;Kim, Song-Yi;Lee, Jue-Hee;Han, Yong-Moon
    • YAKHAK HOEJI
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    • v.55 no.1
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    • pp.39-44
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    • 2011
  • Many reports indicate that $18{\beta}$-glycyrrhetinic acid ($18{\beta}$-GA) from Glycyrrhizae Radix has anti-inflammatory and immunoregulatory activities, whereas reports regarding anticancer activity of the compound are few. In present study, we investigated antitumor effect of $18{\beta}$-GA on tumor caused by A549 cancer cell in mice. Data resulting from the cytotoxicity assay showed that $18{\beta}$-GA caused killing of A549 cells. $LD_{50}$ values of $18{\beta}$-GA were app. 180 ${\mu}M$ and 80 ${\mu}M$, corresponding to 48 hr- and 72 hr-treatments, displaying that the killing activity was more effective as the $18{\beta}$-GA treatment was prolonged. Based on these data, antitumor effect of $18{\beta}$-GA was tested in nude mice. For induction of the tumor, A549 ($3{\times}10^6$ cells/mouse) was injected subcutaneously into the lateral abdomen of nude mice (Balb/c nu/nu). To determine the antitumor effect, nude mice with tumor were given $18{\beta}$-GA (1 mg/200 ${\mu}l$/mouse) intraperitoneally every three days for four times. Tumor-sizes were measured with a caliper for a period of 24 days. Results showed that the $18{\beta}$-GA treatment reduced the tumor-sizes (P<0.05) as compared with negative control nude mice that received diluent (DPBS). The reduction degree was greater than reduction degree by doxorubicin (60 ${\mu}g$/mouse), and the pattern of reduction was almost sustained during the entire period of the observation. In conclusion, our studies demonstrate that $18{\beta}$-GA has antitumor activity to the A549 cancer cell-caused tumor.

Protein Kinases Involved in the Regulation of Wnt/β-catenin Signaling (Wnt/β-catenin 신호를 조절하는 인산화 효소)

  • Shin, Eun-Young;Park, Edmond Changkyun;Hong, Yeonhee;Kim, Gun-Hwa
    • Journal of Life Science
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    • v.23 no.7
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    • pp.947-954
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    • 2013
  • The Wnt/${\beta}$-catenin signaling pathway is an evolutionarily conserved signaling network that is critical for embryonic development and adult tissue maintenance. In addition, aberrant activation of Wnt/${\beta}$-catenin signaling is implicated in the formation of various human diseases, including cancers. Thus, study of the underlying molecular mechanism of Wnt/${\beta}$-catenin signaling regulation is important to understand and treat diseases. Inhibition of aberrant Wnt pathway activity in cancer cell lines efficiently blocks their growth, highlighting the great potential of therapeutics designed to achieve this in cancer patients. Recently, protein kinases have emerged as key regulating components of Wnt/${\beta}$-catenin signaling. In this review, we provide the most recent information on Wnt/${\beta}$-catenin signaling, describe protein kinases involved in Wnt/${\beta}$-catenin signaling, and discuss their potential as drug targets.

A New Coloured Substrate for the Determination of $\beta$-Glucan Degrading Enzyme from Malt and Bacillus subtilis K-4-3 (맥아와 Bacillus subtilis B-4-3의 $\beta$-Glucan 분해 효소측정을 위한 새로운 색소기질)

  • 이성택
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.79-84
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    • 1988
  • Dye materials and cross linking agents were used for the determination of $\beta$-glucanase activities. The objective of this study was to prepare the blue coloured substrates which are sensitive, specific and simple for the determination of $\beta$-glucanase in malt and Bacillus subtilis K-4-3 enzymes. This method is based on the principle of measuring colorimetrically the split product of coloured and cross linked substrate. The best coupling of dye stuff of $\beta$-glucan was cibacron blue 3G-A and the colour released can suitably be measured at 623nm. Optimal concentration of dye and cross linking agents was 1.5g and 1.25$m\ell$ under 0.1N NaOH. The sensitivity comparison proved that the stained $\beta$-glucan method is much more sensitive than the DNS method to determine reducing sugar released by the enzyme.

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Effect of Lipoxygenase, ${\beta}-Carotene$, ${\alpha}-Tocopherol$ and Water Activity on the Oxidation of Linoleic Acid in Starch-Solid Model System (고형상의 모델시스템에 있어서 리놀레산의 산화에 미치는 리폭시게나아제, 카로틴, 토코페롤 및 수분활성의 영향)

  • Kim, Hae-Gyoung;Cheigh, Hong-Sik;Song, Yeong-Ok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.1
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    • pp.23-28
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    • 1992
  • Starch solid model system was employed to investigate the effect of lipoxygenase, ${\beta}-carotene$, ${\alpha}-tocopherol$ and water activity on the oxidation of linoleic acid. The rate of oxidation of linoleic acid by lipoxygenase was increased with the increase in water activity, Addition of ${\beta}-carotene$ and ${\alpha}-tocopherol$ to this system has been shown to inhibit the oxidation of linoleic acid and ${\alpha}-tocopherol$ was more effective antioxidant than ${\beta}-carotene$. However, an increase in the concentration of ${\beta}-carotene$ was found to have a strong antioxidant effect in the solid model system. And also the antioxidative action of ${\beta}-carotene$ was increased with increasing water activity in this system.

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The constituents of taraxacum hallaisanensis roots

  • Yang, Deuk-Suk;Whang, Wan-Kyunn;Kim, Il-Hyuk
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.507-513
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    • 1996
  • Three sesquiterpene lactone compounds, two novel(1.betha.,3.betha.-dihydroxy-6.betha.,11.betha.,4.alpha.,5.alpha.,7.alpha.H -eudesm-12, 6-olide-1-O-.betha.-D-glucopyranoside, 1.betha.,3.betha.-dihydroxy-6.betha.,11.betha.,4.alpha.,5.alpha.,7.alpha.H-eudes m-12,6-olide-1-O-.betha.-D-glucopyranoside) and 1.betha.,3.betha.-dihydroxy-6.betha.,11.betha.,4.alpha.,5.alpha., 7.alpha.H-eudesm-12,6-olide were isolated from the aqueous fraction of MeOH extract of the roots from Taraxacum hallaisanensis (Compositae) employing Amberlite XAD-2, ODS-gel, silica gel and Sephadex LH-20 column chromatographics. Another known compound, (-)-epicatechin, was isolated from the aqueous fraction of the MeOH extract. The total MeOH extract also contained phytosterol and a mixture of .betha.-amyrin acetate, .alpha.-amyrin acetate and lupeol acetate. Structures of isolated compounds were elucidated by spectroscopic parameters of IR, Mass, /sup 13/C-NMR, /sup 1/H-NMR, /sup 1/H-/sup 1/H COSY, /sup 13/C-/sup 1/H COSY and HMBC.

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Biochemical Analysis of Baculovirus-insect Cell Interaction: I. Improved Recombinant ${\beta}-Galactosidase$ Production Using Medium Additives at AcNPV Infection of Insect Cells (Baculovirus-곤충세포 상호반응에 대한 생화학적 연구 -I. AcNPV의 곤충세포 감염시 배지 첨가물을 이용한 재조합 ${\beta}-galactosidase$ 생산 향상-)

  • Lee, Ki-Woong;Kim, Tae-Yong;Chung, In-Sik
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.485-489
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    • 1995
  • The medium additives such as fatty acid, lipid, mannose, folic acid, $CaCl_2$ were examined to enhance recombinant ${\beta}-galactosidase\;({\beta}-gal)$ production in T-flask and air-lift bioreactor. The addition of each component. such as cholesterol, tocopherol, tricaprylin, mannose and folic acid at AcNPV infection of Tn5B1-4 cells enhanced ${\beta}-gal$ production, whereas the addition of $CaCl_2$ did not increase ${\beta}-gal$ production. The recombinant ${\beta}-gal$ production using the infection medium supplemented with a mixture of 0.34 mM cholesterol, 2.2 mM mannose and 0.045 M folic acid was enhanced 2 fold in an air-lift bioreactor, compared to the basal medium.

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Typing of Extended-Spectrum ${\beta}$-Lactamase (ESBL) Producing Enterobacteriaceae Isolated from Slaughterhouse in Pusan, Korea (부산 도축장에서 분리된 광범위 베타 락탐 분해효소(Extended-Spectrum ${\beta}$-Lactamase, ESBL)생성 장내세균의 형별분류)

  • Lee Hun-Ku
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.125-130
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    • 2006
  • The emergence of extended spectrum ${\beta}$-lactamase producing bacteria is causing very serious problems in Korea. Although there have been many reports about these bacteria Isolated from patients and clinical specimens, there is no report of extended spectrum ${\beta}$-lactamase producing organisms isolated from natural environment in Korea. This study was conducted to investigate the biological characteristics and extended spectrum ${\beta}$-lactamase types of eighteen strains of extended spectrum ${\beta}$-lactamase producing Klebsiella pneumoniae and Escherichia coli isolated from a slaughterhouse in Pusan in Korea during 2002 to 2004. Extended spectrum ${\beta}$-lactamases were identified by double-disk synergy test, conjugation, isoelectric focusing values and gene sequencing. Eight strains of Klebsiella pneumoniae and two strains of Eschericha coli were isolated kom pigs and transferred extended spectrum ${\beta}$-lactamase genes to recipient Escherichia coli J53 (sodium azide resistant and ceftazidime senstive) strain by conjugation. The conjugants of extended spectrum ${\beta}$-lactamase genes were alignments and translated to amino acids by BCM and NCBI blast. Eight conjugants of Klebsiella pneumonae were typed TEM-52, and two strains of Escherichia coli, SHV-12, but CMY-1 type were not detected in this study.

Levels and Factors Affecting Serum Carotenoids Concentrations of Adults Living in Rural Area of Korea (한국농촌 성인의 혈청 Carotenoids 농도와 이에 영향을 미치는 요인에 관한 연구)

  • 안윤진
    • Journal of Nutrition and Health
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    • v.30 no.5
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    • pp.489-498
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    • 1997
  • Recent researches suggest that carotenoids are important not only as provitamin A but also for prevention of chronic diseases. This study was conduction to determine levels and factors affecting serum levels of lutein + zeaxanthin, $\beta$-cryptoxanthin, and $\beta$-carotene in 93 adults living in rural area of Korea. Fasting blood samples were collected and serum carotenoid levels were measured by HPLC. Dietary intake was estimated by 24 hour recall method and frequency questionnare of major food groups. Mean serum concentration of lutein + zeaxanthin was 616.32 nmol/L, $\beta$-cryptoxanthin was 856.95nmol/L, and $\beta$-carotene was 242.90nmol/L. Serum $\beta$-carotene levels in study subjects were very low. Both $\beta$-cryptxanthin and $\beta$-carotene were negatively correlated with serum triglyceride and positively correlated with total-choesterol and LDL-cholesterol. Serum levels of female subjects were significantly higher than males in all carotenoids. For age groups, subjects in their 30's were shown to have the highest concentration of all carotenoids. Lutein + zeaxanthin were lowest in subjects in theri 40's while $\beta$-crytoxanthin and $\beta$-carotene levels were lowest in subjects in their 60's. The $\beta$-carotene levels in non-smokers were significantly higher than in drinkers. Lutein+zeaxanthin levels were significantly higher among subjects consuming more green and yellow vegetables by frequency questionnarie. In conclusion, serum carotenoids were affected by sex, age, serum lipids, smoking, and alcohol intake. Intake of vegetables and fruits could affect by sex, serum lipids, smoking, and alchol intake. Intake of vegetables and fruits could affect serum lutein+zeaxanthin level. This data indicated that compared to other studies, Korean adults in rural areas have high lutein+zeaxanthin concentratins and low $\beta$-carotene concentrations in serum. High lutein+zeaxanthin levels may be related to high consumption of vegetables in these subjects.

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