• Title/Summary/Keyword: $P388D_1\

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Apoptosis Induced by Adenosine 5'-triphosphate in Mouse Leukemic Cells (Mouse Leukemia 세포에서 Adenosine 5'-triphosphate에 의한 Apoptosis)

  • Joo, Nan-Young;Park, Kyu-Sang;Chung, Hae-Sook;Kong, In-Deok;Lee, Joong-Woo
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.6
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    • pp.817-824
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    • 1997
  • Extracellular ATP elicits various biological responses and plays a significant role in physiological regulation. Recently, ATP-induced growth inhibitions were reported in some tumor cell lines, but these effects and mechanisms are not well hewn. This study was conducted to investigate ATP-induced growth inhibition in mouse $leukemic(P388D_1)$ cells. ATP inhibited cell growth in a dose-dependent manner as analyzed by MTS assay$(IC_{50}: 33.1\;{\mu}M)$. Nucleotides other than ATP, such as ADP$(37.5;{\mu}M)$ and AMP$(33.2;{\mu}M)$ had the same effects as ATP but adenosine$(57.8;{\mu}M)$ showed less effect than ATP. ATP attenuated the cells in $G_0/G_l\;and\;G_2/M$ phases but increased those in S phase in flow cytometric analysis. Hypodiploid cells$(A_0)$, the presumptive findings of apoptosis, were found among the ATP-treated cells. ATP induced DNA fragmentation into $180{\mu}200\;bps $as measured by electrophoresis. some apoptotic cells were stained by TUNEL method. ATP increased the intracellular free $Ca^{++}$concentration$([Ca^{++}]_i)$ and the increment of $([Ca^{++}]_i)$ was caused by influx from the extracellular space. These results suggest that extracellular ATP induces growth inhibition through apoptosis.

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Studies on the cytotoxicity and Antimicrobial Effects of the Extract of Houttuynia cordata (IV) (어성초 추출물의 세포독성과 향균효과 (IV))

  • Lee Jeong-Ho;Park Nang-Kyu;Yang Eun-Yeong;Lee Hyun-Ok;Han Dong-Min;Baek Seung-Hwa
    • Journal of Society of Preventive Korean Medicine
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    • v.4 no.1
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    • pp.144-151
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    • 2000
  • This study was carried out to evaluate cytotoxic effects of Houttuynia cordata Thunberg extracts on murine leukemia tumor cell lines. Disruptions in cell organelles were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazol iumbromide (MMT) assay. The comparison of $IC_{50}$ values of Houttuynia cordata Thunberg extracts on $L1210,\;P388D_1$ and Vero cell lines showed that the methanol extract of Houttuynia cordata Thunberg indicated the most antitumor activity in the MTT assay. In order to develop a antimicrobial agent, dried Houttuynia cordata Thunberg was extracted with several solvents, and then antimicrobial activity was investigated. The minimal inhibitory concentration (MIC) of the extracted substance against microorganisms were also examined. Antimicrobial activity of amocla and ketoconazole as references was compared to those of other solvent extracts such as $H_2O$, n-hexane, chloroform, ethyl acetate ethanol and methanol. The antimicrobial activity of all extracts from the sample had growth inhibition activity against gram-negative bacteria, yam-positive bacteria and fungi $(MIC,\;>\;200\;{\mu} g/ml)$. These results suggest that the methanol soluble extract of Houttuynia cordata Thunberg may be a valuable choice for the studies on the treaeent of murine leukemia tumor cell lines and antimicrobial agents.

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X-ray Crystal Structure and Luminescence Properties of Pd(II) and Pt(II) Complexes with Dithiopyrrole

  • Kang, Jun-Gill;Cho, Dong-Hee;Park, Changmoon;Kang, Sung Kwon;Kim, In Tae;Lee, Sang-Woo;Lee, Ha-Hyeong;Lee, Young-Nam;Lim, Dae-Won;Lee, Sung-Jae;Kim, Sung-Ho;Bae, Young-Ju
    • Bulletin of the Korean Chemical Society
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    • v.29 no.3
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    • pp.599-603
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    • 2008
  • The complexes Pd(nbmtp)Cl2 and Pt(nbmtp)Cl2 (nbmptp = 1-nonyl-3,4-bis(methylthio)pyrrole) were prepared and their x-ray structures were determined at room temperature. The four-coordinated metal unit and the pyrrole ring formed a nearly planar geometry. The free ligand dissolved in CH2Cl2 produced two luminescence bands associated with the lone-pair electron of S (l max = 525 nm) and the pyrrole p electron (l max = 388 nm). When the two complexes were dissolved in CH2Cl2, these two luminescence bands were also observed, although the low-energy band was blueshifted. For the crystalline Pt(II) complex, only the strong charge transfer band (l max = 618 nm) from the d* orbital of Pt resulted from excitation of the lone-pair electron of S.

Immunoadjuvant Activity of Korean Mistletoe Lectin B-chain (한국산 겨우살이 Lectin B-chain의 면역증강 효과)

  • Her, Sun-Mi;An, Hyo-Sun;Kim, Kyu-Dae;Kim, Young-Hoon;Kim, In-Bo;Yoon, Taek-Joon;Kim, Jong-Bae
    • Korean Journal of Pharmacognosy
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    • v.42 no.3
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    • pp.246-252
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    • 2011
  • Korean mistletoe Lectin (KML-C) is composed of A and B sub-chain. B chain binds to carbohydrates on cell surface and A chain hinders translation and induces an apoptosis as a RIP (ribosome inactivating protein). KML-C has very strong biological activities, it has seriously limits to use as a cancer therapy or adjuvant because of its toxicity to normal cells. This study is therefore conducted to see if B chain of KML-C might have immunological activity, especially adjuvant activities with less toxicity. We isolated B chain from KML-C using the lactose affinity chromatography, and examined their immunoadjuvant activity. The isolated B-chain did not show any cytotoxicity against tumor cell, RAW264.7, and P388D1 while KML-C had a very strong toxicity. This non-toxic effect was observed also by in-vivo study. Both humoral and cellular immunities were observed ; the antibody titer was increased when the mice were immunized with B-chain used as adjuvant like Freund's adjuvant, indicating that B chain of mistletoe lectin alone might be used for adjuvant; it also increased DTH in cellular immunity. These results suggest that B-chain of KML-C might be used for adjuvant used for the production of antibody or vaccine with less toxicity.

Growth Response of Strawberry Seedlings to Application of Solid Chlorella (고형 클로렐라 시비에 대한 딸기유묘의 생육 반응)

  • Young-Nam Kim;Jun Hyeok Choi;Hyeonji Choe;Keum-Ah Lee;Young-Eun Yoon;Vimalraj Kantharaj;Yong Bok Lee
    • Korean Journal of Environmental Agriculture
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    • v.42 no.4
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    • pp.382-388
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    • 2023
  • A liquid-type chlorella (LC) produced by self-cultivation is used for strawberry cultivation by farmers in Korea. This study aimed to investigate seedling growth in two strawberry cultivars 'Soraya' and 'Haruhi' after applying solid-type chlorella (SC) in the greenhouse for 30 d. The treatments were as follows: Control, LC (0.2% of 1.0 × 107 cells/mL), ¼ SC (12.5 g/m2), ½ SC (25 g/m2), and SC (50 g/m2). Compared to the control, in the ½ SC treatment, dry weight of the seedlings increased at 30 days after treatment (DAT). Phosphorus content in the leaves of 'Soraya' seedlings of the SC treatment increased compared to that of the control. At 10 and 30 DAT, photosynthetic pigments including chlorophylls and carotenoids in the seedlings of both cultivars increased with the ½ SC treatment. These results indicate that the application of SC improved the growth of strawberry seedlings and could replace LC.

Effects of Microbacterium laevaniformans Levans Molecular Weight on Cytotoxicity

  • Oh, Im-Kyung;Yoo, Sang-Ho;Bae, In-Young;Cha, Jae-Ho;Lee, Hyeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.985-990
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    • 2004
  • Levans produced from Microbacterium laevaniformans were isolated, characterized, and fractionated by molecular weight. TLC, HPLC, and GC-MS analyses of the exopolysaccharide showed that it was a fructan-type polymer and was composed of (2,6)- and (2,1)-glycosidic linkages. $^{13}C$-NMR analysis proved that the polysaccharide was mainly a $\beta$-(2,6)-linked levan-type polysaccharide. To investigate the cytotoxicity of the acetone-precipitated levan fractions such as M1, M2, and M3, HepG2, P388D1, U937, SNU-1, and SNUC2A cell lines were screened. Among the cell lines tested, the cytotoxicity of M1- M3 fractions were detected from only SNU-1 and HepG2 cells at the dosage level of $100-800\mu\textrm{g}ml$. The M2 fraction M_r$, 80,000) at 400 $mu{g/ml}$ had the greatest cell growth inhibition (84.6%) on SNU-1, while the M1 $(M_r$, 50,000) at $800\mu\textrm{g}ml$ showed the greatest (46.32%) on HepG2. To obtain more uniform M_r$ fractions of levan, the levan was further fractionated from S1 $(M_r$ 1,000,000) to S5 $(M_r$ 10,000) using gel permeation chromatography. Again, the S1-S5 fractions had strong cytotoxicity on SNU-1 and HepG2 cell lines. The greatest inhibition effects of S4 $(M_r$ 80,000) on SNU-1 and S5 $(M_r$ 10,000) on HepG2 were shown to be 49.5% and 73.0%, respectively. The cytotoxicity of the levan fractions was more effective on SNU-1 than on HepG2. Although the relationship between the Mw and the cytotoxicity was not clear, smaller $M_r$, fractions of levan showed greater growth inhibition effect on the cancer cell lines in general. Therefore, it was indicated that a specific Mw class of levan is responsible for the effective cytotoxicity.

Strain Development for the Over-production of Alkaline Protease from Vibrio metschnikovii by Molecular Evolution (분자진화 기술을 통한 Vibrio metschnikovii 유래 고활성 알칼리성 단백질 분해효소 생산균주 개발)

  • Shin, Yong-Uk;Lee, Gwa-Soo;Jo, Jae-Hyung;Lee, Hyune-Hwan
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.383-388
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    • 2010
  • Alkaline protease-overproducing strains of Vibrio metschnikovii were developed by using the molecular evolution from the classical mutants V. metschnikovii L12-23, N4-8, and KS1. Each vapK (Vibrio alkaline protease K) was obtained from the genomic DNAs of mutants by PCR to carry out the DNA shuffling. The modified vapK-1 obtained by DNA shuffling was used again as a template for the error-prone PCR to make the vapK-2. Both genes were cloned in the plasmid pKF3 to construct the recombinant plasmids which have one or two copies of the modified genes. The recombinant plasmids were back-transformed to V. metschnikovii KS1 to construct recombinant V. metschnikovii that expresses the alkaline protease. About 3.9-fold more protease activity was measured in the strain which has the plasmid containing two copies of vapK-2 when compared to strain KS1. When compared to wild type V. metschnikovii RH530, 43-fold more activity was achieved. Comparison of amino acids among vapK, vapK-1, and vapK-2 revealed that the active sites was highly conserved and not changed. However, many amino acids except the active sites were changed. These results suggested that the changes in amino acids might play an important role in the increase of protease activity by allowing the easy access of substrate to active sites of the protease. The fermentation of alkaline protease from the V. metschnikovii KS1 harboring the plasmid that contains two copies of vapK-1 showed the possibility of this strain to be used as industrial producer.

The Study of DNA Mutations of Phenylketonuria in Koreans (한국인에서의 페닐케톤뇨증의 유전자변이에 대한 고찰)

  • Yoo, Su-Jung;Hong, Yong-Hee;Lee, Yong-Wha;Jung, Sung-Chul;Ki, Chang-Seok;Lee, Dong-Hwan
    • Journal of Genetic Medicine
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    • v.5 no.1
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    • pp.26-33
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    • 2008
  • Purpose : Phenylketonuria(PKU) is an inborn error of metabolism and a genetic disorder resulting from a deficiency of phenylalanine hydroxylase(PAH) and decreased activity of tetrahydrobiopterin(BH4).In this study the correlation between the DNA mutation and clinical manifestations was investigated and PAH DNA mutations were compared bewteen Asian and Caucasian populations. Methods : DNA was isolated from peripheral leukocytes. The PAH gene was amplified by Polymerase Chain Reaction(PCR) and the sequence was analyzed with Multiplex Ligation-dependent Probe Amplification(MLPA). Results : We characterized the PAH gene of 102 independent Korean patients with PKU. PAH nucleotide sequence analysis revealed 44 different mutations, including 10 novel mutations comprising 9 missense mutations(N207D, K95del, A447P, G344D, P69S, S391I, A202T, G103S, and I306L) and 1 novel splice-site variant mutation(IVS10-3C>G). R243Q was the most prevalent mutation in this study. A259T has not previously been reported in Asian populations, but we found that this mutation had a frequency of 10.1% in our study. Furthermore, the genotypes of $BH_4$ responsive patients were analyzed and were divided into two groups: $BH_4$ medication-only group and $BH_4$ medication with diet therapy group. In the $BH_4$ medication-only group and $BH_4$ medication with diet therapy group, R241C was the most common mutation. Conclusion : Novel mutations in the PAH gene of PKU patients are still being discovered. Additional information as to the frequency of mutations in the tetrahydrobiopterine responsive gene is also accumulating. We anticipate that knowledge of these PKU gene mutations will assist the diagnosis, genetic counseling, and therapeutic treatment of PKU patients in future.

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Community Structure and Distribution of Natural Seaweed Beds on the Eastern Coast of Korea (동해안 천연 해조장의 군집구조와 분포 특성)

  • Park, Gyu Jin;Ju, Hyun;Choi, Ok In;Choi, Chang Geun
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.23 no.4
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    • pp.338-346
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    • 2017
  • Natural seaweed beds and habitat environments were investigated using quantitative and qualitative methods from May to December 2015 at 3 sites in Gangneung, Uljin, and Busan along the eastern coast of Korea. In total, 9 green, 23 brown, and 64 red algal taxa were identified. The biomass of the seaweed at Gangneung was 173.2 to $613.8wet\;wt.g/m^2$ of Dictyota divaricata, 360.8 to $520.4wet\;wt.g/m^2$ of Symphyocladia linearis, and 25.9 to $470.8wet\;wt.g/m^2$ of Undaria pinnatifida. At Uljin, these numbers were 5.5 to $256.2wet\;wt.g/m^2$ of Plocamium telfarirae and 46.8 to $241.5wet\;wt.g/m^2$ of Agarum clathratum. The biomass of Sargassum coreanum and Ecklonia cava were 388.1 to $6,972.4wet\;wt.g/m^2$ and 194.9 to $958.5wet\;wt.g/m^2$, respectively, at Busan. S. coreanum and E. cava showed higher biomass compared to other seaweed at Busan. The biomass rate represented an average of 19.2 percent of the total population, ranging from 0.0 to 55.5 percent in Gangneung. In Uljin, the average was calculated as 63.8 percent, and this figure was 48.5 percent in Busan. The percentage of barren ground averaged 46.7 percent in Gangneung and 91.1 percent in Uljin. Uljin showed the highest percentage of barren ground compared to other regions. Sea urchin density appeared to be $6.0ind./m^2$ in Gangneung, $7.0ind./m^2$ in Uljin, and $2.0ind./m^2$ in Busan, with the lowest sea urchin density being that of Busan. In conclusion, the composition of species, appearance ratio, and abundance of vegetation found were similar to previous studies, but it is thought that continuous monitoring is needed due to concerns about physical and chemical pollution caused by global warming, climate change, and coastal development.

Antioxidant Activities of Rubus coreanus Miquel and Morus alba L. Fruits (복분자와 오디의 항산화 특성)

  • Jun, Hyun-Il;Kim, Young-Ae;Kim, Young-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.381-388
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    • 2014
  • The antioxidant activities of extracts from Rubus coreanus Miquel (black raspberry) and Morus alba L. (mulberry) fruits were investigated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, 2,2-azino-bis-(3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS) assay, and reducing power assay. Aqueous mixtures of ethanol, methanol, and acetone were analyzed in order to determine the most effective extraction solvent for the two fruits. Black raspberry and mulberry extracts with the 60:40 acetone-water mixtures (v/v) showed the highest DPPH radical scavenging activities (56.2 and 85.2%, respectively) compared to the other extraction solvents. The 60% acetone extract was finally selected as an extraction solvent and then sequentially fractionated according to solvent polarity. Among the fractions of the two fruits, the ethyl acetate fraction showed the highest antioxidant activity as well as total phenolic and total flavonoid contents. In addition, there were high correlation coefficients between antioxidant activities and their contents. The $EC_{50}$ value of the ethyl acetate fraction from mulberry fruit was 2.2 times lower than that of butylated hydroxytoluene (BHT) in DPPH assay. The major phenolic acid and anthocyanin of the two fruits were protocatechuic acid and cyanidin-3-glucoside, respectively.