• 제목/요약/키워드: IIE

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Purification and Partial Amino Acid Sequence of a Bacteriocin Produced by Lactococcus, sp. HY449 (Lactococcus sp. HY449가 생산한 Bacterisocin의 정제)

  • 오세종;이상준;김경태;김상교;박연희;백영진
    • Microbiology and Biotechnology Letters
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    • v.29 no.3
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    • pp.155-161
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    • 2001
  • A bactriocin produced by Lactrococcus sp. HY449 was purified by sequential purfication steps such as n-propanol-acetone precipitation ion -exchange chromatography using CM-Sequential CL6B. gel filtration chromatography using Sephacry HR100 and reverse-phase chromatography using pro RPC HR 5/10. Reverse-phase chromatography the final step of the purfication yielded a single symmetrical peak of bacteriocin activity The purification resulted in final yield of 3.25% and 413.35 fold increase of the specific activity of bacteriocin. The active fraction from reverse-phase chromatography was used for N-terminal amino acid analysis . The purified bacteriocin contained isoleucine, leucine, methionine, and glycine at but N-terminal end no aromatic amino acids. Calculation of the number of amino acid residues in the bacteriocin revealed that it is consisted of 32 residues assuming the molecular weight of bacteriocin to be about 3.6kDa. Edman degrandation elucidated amino acid residues of the first four of the N-terminus to be $NH_2$-Ile-Leu-Pro-GIn.

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Role of PI3-kinase and MAP Kinases in the ARE-mediated Glutathione S-Transferase Induction by Phytochemicals: Comparison with the Oxidative Stress Caused by Decreased Glutathione

  • Kim, Sang-Geon;Kang, Keon-Wook
    • Toxicological Research
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    • v.17
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    • pp.251-256
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    • 2001
  • The expression of phase II detoxifying enzymes is affected by a variety of compounds and the induction of the enzymes plays an essential role in chemoprevention. A variety of phytochemicals such as sulfur-containing chemoprotective agents (SCC) may trigger cellular signals and activate phase II gene expression through ARE activation. see induces glutathione S-transferases. Studies were conducted to investigate the role of mitogen-activated protein (MAP) kinase and phosphatidylinositol 3-kinase (PI3-kinase) in the induction of GST (e.g. rGSTA2) by sec. We also studied the MAP kinase pathway responsible for the GST expression by see and compared that with the pathway activated by oxidative stress as a result of sulfur amino acids deprivation (SAAD). see inhibited phosphorylation of ERK1/2 although the effect of see on JNK and p38 MAP kinase was minimal. Wortmannin and LY294002. PI3-kinase inhibitors. abolished the increases in rGSTA2 mRNA and protein levels by SCC. Deprivation of cystine and methionine caused oxidative stress in H4IIE cells. as evidenced by a decrease in the reduced glutathione and an increase in prooxidant production. Electrophoretic mobility shift assay revealed that the ARE complex consisting of Nrf-1/2 and Maf proteins was activated 12~48 h. The rGSTA2 mRNA and protein levels were increased by SAAD. Activation of ARE and induction of rGSTA2 were both completely inhibited by PI3-kinase inhibitors. Inhibition of p38 MAP kinase by SB203580 prevented the ARE-mediated rGSTA2 induction. The results of this study showed that PI3-kinase might play an essential role in the ARE-mediated rGSTA2 induction by see or SAAD and that the dual MAP kinase pathways were responsible for the enzyme induction.

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Genotoxic Effect of Air-borne Particulate Matter in Residential Area of Seoul City (서울시내 주거지역 미세먼지의 유전독성 영향)

  • Oh Seung Min;Sung Hye Kyoung;Kim Eun Sil;Kim Jong Geuk;Ryu Byung Taek;Chung Kyu Hyuck
    • Environmental Analysis Health and Toxicology
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    • v.20 no.4 s.51
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    • pp.365-374
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    • 2005
  • Ambient air particulate matters are classified into two distinct modes in sire distribution, namely the coarse and fine particles. Correlation between high particulate concentration and adverse effect on human populations has long been recognized. However, the toxicology of these adverse efforts has not been clarified. We investigated the genotoxic effect of PM 2.5 collected from urban area in Seoul by comet assay (A549 cells), CBMN assay (CHO-K1 cells) and EROD-microbioassay (H4IIE cells). Results from in vitro micronucleus assay and comet assay showed that PM 2.5 samples collected from traffic area, residential area and indoor air induced chromosomal damage and DNA breakage in a non-cytotoxic dose. The complex mixture effect of these PM 2.5 extracts was quantified by EROD-microbioassay in terms of its bio-TEQ (biologiral -TCDD equivalent concentration) which was 70.87$\pm$28.07, 93.55$\pm$21.80 and 14.31 $\pm$ 1.10 ng/g-PM 2.5 in traffic area, residental area and indoor air samples, respectively. Conclusively, we suggested that PM 2.5 collected from traffic area and residential area contains CYPIA inducer and genotoxic materials.

Evaluation of Antioxidant Activity of Platycodon grandiflorum (장생도라지 (Platycodon grandiflorum)의 항산화 활성 평가)

  • Kim, Chung-Hyeon;Jung, Bong-Yong;Jung, Sun-Ki;Lee, Chul-Ho;Lee, Hyun-Sun;Kim, Bong-Hee;Kim, Sang-Kyum
    • Environmental Analysis Health and Toxicology
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    • v.25 no.1
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    • pp.85-94
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    • 2010
  • Platycodon grandiflorum, Doraji as Korean name, is one of the most widely used traditional oriental medicine for bronchial diseases and also used as a folk remedy for geriatric diseases and inflammatory diseases. In recent studies, it has been reported that some effect of P. grandiflorum is derived from its antioxidant activity, although there is still a lack of evidence to establish its oxy-radical scavenging activity. In this study, total oxy-radical scavenging capacity (TOSC) assay was used to evaluate antioxidant activity of total extracts (T-PG), polysaccharide fraction (Po-PG), and saponin fraction (Sa-PG) isolated from P. grandiflorum against peroxyl radicals and peroxynitrites. And MTT assay was taken to assess cyto-protective effects of T-PG, Po-PG and Sa-PG in H4IIE cells treated with hydrogen peroxide and tert-butylhydroperoxide. In the TOSC assay, Sa-PG showed strong oxy-radical scavenging capacity compared with T-PG and Po-PG. In cell-based assay, T-PG and Po-PG protected cells from oxidative stress, but Sa-PG did not protect cells because of cytotoxicity of Sa-PG. These results suggest that the saponin components of P. grandiflorum have relatively strong antioxidant capacity and cytotoxicity in rat hepatoma cells.

Glutathione S-transferase T1, M1 and P1 Genetic Polymorphisms and Susceptibility to Colorectal Cancer in Turkey

  • Gorukmez, Ozlem;Yakut, Tahsin;Gorukmez, Orhan;Sag, Sebnem Ozemri;Topak, Ali;Sahinturk, Serdar;Kanat, Ozkan
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.3855-3859
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    • 2016
  • Colorectal cancer (CRC) is reproted to be the third most common cancer worldwide and the fourth most common cause of cancer related deaths. CRC is considered to be a multifactorial disease whose risk varies due to the complex interaction between individual genetic basis and disposure to multiple endogenous factors. Glutathione S-transferases are pro-carcinogenic in CRC and are required for the conjugation between chemotherapeutics and broad spectrum xenobiotics. One hundred and eleven patients with CRC and 128 control subjects without any cancer history were enrolled in this study. Multiplex PCR was applied to determine polymorphisms for the GSTT1 and M1 genes, and PCR-RFLP was applied for the GSTP1 (Ile105Val) gene polymorphism. Values p<0.05 were defined as statistically significant. We detected a significant high correlation between predisposition for CRC and presence of the Ile/Ile genotype of the GSTP1 (IIe105Val) gene polymorphism, but we did not find a significant relationship between predisposition for CRC and GSTT1 and M1 deletion polymorphisms. In addition, we did not determine a relationship between GSTT1, M1 and P1 gene polymorphisms and any clinicopathological features of CRC. GSTT1 null/GSTM1 positive and GSTT1 null/GSTM1 positive/GSTP1 Ile/Ile genotypes were significantly higher in the patient group. Our results revealed that there is no relationship among CRC, its clinicopathologic features, and GSTT1 M1 gene polymorphisms. However, there was a significant correlation between CRC and the GSTP1 Ile/Ile genotype. Further studies with larger patient groups are required to delineate the relationships between GST gene polymorphisms and the clinicopathologic features of CRC in Turkey.

Synthesis of Silicon-Carbon by Polyaniline Coating and Electrochemical Properties of the Si-C|Li Cell

  • Doh, Chil-Hoon;Kim, Seong Il;Jeong, Ki-Young;Jin, Bong-Soo;An, Kay Hyeok;Min, Byung Chul;Moon, Seong-In;Yun, Mun-Soo
    • Bulletin of the Korean Chemical Society
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    • v.27 no.8
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    • pp.1175-1180
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    • 2006
  • Si-C composites were prepared by the carbonization of polyaniline (PAn) coated on silicone powder. The physical and electrochemical properties of the Si-C composites were characterized by particle-size analysis, X-ray diffraction, scanning electron microscopy, and battery electrochemical tests. The average particle size of Si was increased by the coating of Pan but somewhat reduced by the carbonization to give silicone-carbon composites. The co-existence of crystalline silicone and amorphous-like carbon was confirmed by XRD analyses. SEM photos showed that the silicone particles were well covered with carbonaceous materials, depending on the PAn content. Si-C$\mid$Li cells were fabricated using the Si-C composites and tested using galvanostatic charge-discharge. Si-C$\mid$Li cells gave better electrochemical properties than Si|Li cells. Si-C$\mid$Li cells using Si-C from HCl-undoped precursor PAn showed better electrochemical properties than precursor PAn doped in HCl. The addition of an electrolyte containing 4-fluoroethylene carbonate (FEC) increased the initial discharge capacity. Also, another electrochemical test, the galvanostatic charge-discharge test with GISOC (gradual increasing of the state of charge) was carried out. Si-C(Si:PAn = 50:50 wt. ratio)|Li cell showed 414 mAh/g of reversible specific capacity, 75.7% of IIE (initial intercalation efficiency), 35.4 mAh/g of IICs (surface irreversible specific capacity).

Production of monoclonal antibodies specific to the surface antigens of chicken peripheral blood mononuclear cells (닭의 혈액내 단핵세포 표면항원 특이 단클론성 항체 생산)

  • Choi, Jun-Gu;Sung, Haan-Woo;Kim, Sun-Joong
    • Korean Journal of Veterinary Research
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    • v.42 no.2
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    • pp.209-217
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    • 2002
  • This study was performed to produce monoclonal antibodies (mAb) specifically reacting with chicken leukocyte surface antigens. Popliteal lymph node cells of BALB/c mice previously immunized through foot-pad with peripheral blood mononuclear cells (PBMC) of chickens separated by Ficoll-Histopaque method. They were fused with P3X63Ag14 mouse myeloma cells. A total of 34 hybridomas secreted antibodies specifically binding to the PBMC. According to the reactivity patterns with PBMC, the mAbs were divided into 4 groups. Group 1 mAbs (IIB3, IIB10, IIE10) specifically reacted with non-adherent lymphocytes but not with adherent cells which were mainly composed of thrombocytes and monocytes in PBMC culture. These mAbs were reactive with 25-59% of thymus cells and 42-64% of spleen cells of chickens. They did not show any significant reactivity with cells in the bursa of Fabricius, T-cell (MDCC-MSB1) and B-cell (LSCC-1104B1) lines. These results indicate that Group I mAbs specifically reacted with T-lymphocyte subpopulation. Monoclonal antibodies in Group II (IC6, IG2-2 and IID9) showed specific reactivity with monocytes but not with thrombocytes or non-adherent cells in PBMC culture. These mAbs, though not reacted with the chicken macrophage cell line, HD11, also bound to macrophages of the spleen and lung in immunohistochemical staining. Five mAbs in Group III showed characteristics of binding to lymphocytes and monocytes, but not to thrombocytes. Twenty-three mAbs in Group IV showed specific reactivity to lymphocytes, monocytes, and thrombocytes. Two mAbs (IC3 and IE9) in Group IV reacted with most of PBMC.

Methods for Evaluation of Antioxidant Activity: Application to Taurine (항산화 활성의 평가를 위한 연구법: 타우린의 적용)

  • Kim, Bong-Hee;Oh, Jung-Min;Yun, Kang-Uk;Kim, Chung-Hyeon;Kim, Sang-Kyum
    • Toxicological Research
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    • v.23 no.3
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    • pp.263-269
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    • 2007
  • Although taurine (2-aminoethanesulfonic acid) can inhibit oxidative stress in both animal and epidemiological studies, it is obscure whether taurine directly scavenges oxy-radicals or indirectly regulates oxidant production and/or antioxidant defense system. The reason for this discrepancy remains unknown but may be due, in part, to the lack of a validated assay system for evaluating oxy-radical scavenging capacity. The antioxidant activities of taurine and hypotaurine (2-aminoethanesulfinic acid), a precursor of taurine, against peroxyl radicals, hydroxyl radicals and peroxynitrites were determined by the total oxy-radical scavenging capacity (TOSC) assay and cell-based assay using H4IIE cells. tert-Butylhydroperoxide or hydrogen peroxide-induced cell toxicity determined by MTT assay was markedly inhibited by 10mM taurine or hypotaurine. The tert-butylhydroperoxide- or hydrogen peroxide-induced changes in oxidative stress markers, such as cellular glutathione and malondialdehyde, were ameliorated by 10mM taurine or hypotaurine. However, specific TOSC values calculated from the slope of the linear regression for taurine against peroxyl radicals, hydroxyl radicals or peroxynitrites were all less than 1 TOSC/mM. On the other hand specific TOSC values for hypotaurine against peroxyl radicals, hydroxyl radicals or peroxynitrites were 48, 2096, or 69 TOSC/mM, respectively. These results suggest that taurine protects cells against oxidative insults, which is not ascribed to directly scavenging activity of taurine against oxy-radicals. These results support the idea that the oxidation state of sulfur in antioxidants may be a determinant of oxy-radical scavenging capacity.

The Effect of Salivary Gland of Streptozotocin Induced Diabetic Rats by Stress (스트레스가 Streptozotocin유도 당뇨백서의 타액선에 미치는 영향)

  • 김홍모;전양현;홍정표
    • Journal of Oral Medicine and Pain
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    • v.22 no.1
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    • pp.65-80
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    • 1997
  • Cytochrome P450 is an oxidase involved in oxidation of alcohol and is known to be an activator of carcinogen. The present study was perfomed to analyze the effect of diabetes and cold stress on the expression of Cytochrome P450 IIE1(CYPIIE1) in the liver and salivary glands in rats by an immunoblot analysis. Sixty three divided into 4 groups; 1) 20 rats belonging to group I were allowed diabetes (40mg/kg. I.V.) 2) 20 rats of group II were bathed in cold water for 30 seconds twice a day 3) 20 rats comprising group III were received diabetes and cold stress as described above 4) 3 rats of group IV were selected as a control. The rats were sacrificed at the end of the same day 1, 2, 3, 4 weeks experiment. The liver and parotid glands were removed and stored at $-70^{\circ}C$ until use. The stored organs were homogenized for 10 seconds and the supernatants were obtained by centrifugation. The proteins of the supernatants were separated by sodium dodecyl sulfate- polyacrylamide gel electrophoresis and subjected to Western blotting. The blotted membranes were incubated with polyclonal antibodies to CYPIIE1. And sepimens were observed with light microscope also under the Hematoxillin-Eosin staining. The obtained results were as follows : 1. In diabetes group, acini had changed to degeneration severely 1 week experiment, but repaired gradually in lapse of time. 2. In diabetes group, septal connective tissue had changed to degeneration little by little from 1 week after experiment, and progressed severely in lapse of time. 3. In stress group, acini had not changed remarkably, but slightly separated each other 3 weeks after experiment. 4. In diabetes and stress group, histological feature had changed remarkably campare with in the group of diabetes only. 5. In all experimental group, CTPIIE1 had expressed remarkably in the liver tissue, but not in the parotid gland tissues. 6. In diabetes and stress group, CYPIIE1 had expressed remarkably campare with in the group of diabetes only.

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Antioxidant capacity and hepatoprotective effect of ethyl acetate fraction from shoot of Aralia elata on alcohol-induced cytotoxicity (두릅 아세트산 에틸 분획물의 산화방지 효과 및 알코올에 대한 간세포 보호효과)

  • Kwon, Bong Seok;Park, Seon Kyeong;Kim, Jong Min;Kang, Jin Yong;Park, Sang Hyun;Kang, Jeong Eun;Lee, Chang Jun;Park, Su Bin;Yoo, Seul Ki;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.50 no.2
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    • pp.216-224
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    • 2018
  • To evaluate physiological effect of Aralia elata, in vitro antioxidant activity and hepatic protective effects were investigated. Ethyl acetate fraction from Aralia elata (EFAE) had higher total phenolic content than other fractions (n-hexane, chloroform, and distilled water layers). EFAE also showed significantly greater radical scavenging activity against 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH), than other fractions. Moreover, EFAE showed dose-dependent inhibitory effect of malondialdehyde (MDA). Hepatoprotective effects of EFAE against ethanol- and $H_2O_2$-induced oxidative stress and cytotoxicity in H4IIE and HepG2 hepatic cells were examined using 2',7'-dichlorofluorescein diacetate (DCF-DA) and 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. The results showed that EFAE reduced cellular oxidative stress, and increased hepatic cell viability. In addition, EFAE inhibited ethanol-induced lipid accumulation in HepG2 cells. Finally, physiological substances of EFAE were analyzed using high performance liquid chromatography (HPLC), and the major bioactive compounds identified were 3,5-dicaffeoylquinic acid and chlorogenic acid.