• Title/Summary/Keyword: AP-l

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Detection of Ref-1 (Redox factor-1) Interacting Protein Using the Yeast Two-hybrid System (Yeast two-hybrid system을 이용한 Ref-1 (redox factor-1) 결합 단백질의 분리 및 동정)

  • 이수복;김규원;배문경;배명호;정주원;안미영;김영진
    • Journal of Life Science
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    • v.14 no.1
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    • pp.26-31
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    • 2004
  • Redox factor-1 (Ref-1), known as a redox regulator, controls the DNA binding of AP-1 and is activated in HT29 colon cancer cells by hypoxia in vitro. REF-1 also increases tile DNA binding affinity of Hypoxia-inducible Factor-lalpha$ (HIF-lalpha$), HIF-like Factor (HLF) and early growth response-1 (Egr-1) which induce expression of the genes involved in angiogenesis, so that we speculate that REF-1 may play a role in hypoxia-induced angiogenesis. In this research we tried to detect novel proteins interacting with REF-1 using Yeast two-hybrid system using full-length REF-1 cDNA as bait. As result of such screening we detected 3 positive clones. DNA sequencing and GeneBank search revealed that one of the clones contained the same sequences as M.musculus cDNA for tioredoxin.

Fermentation of Environmental Friend Total Mixed Ration and Alteration of Rumen Fermentation Characteristics (환경친화적 섬유질 배합사료의 발효와 반추위 발효특성 변화)

  • Ryu, Chae-Hwa;Park, Myung-Sun;Park, Chul;Choi, Nag-Jin;Cho, Sang-Buem
    • Korean Journal of Organic Agriculture
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    • v.25 no.2
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    • pp.461-473
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    • 2017
  • Total mixed ration (TMR) including concentrate diet and roughage together have been used for the ruminant animal. Relatively high concentrations of moisture and water soluble carbohydrate are representative feature of TMR. Those moisture and water can also provide a niche for bacterial growth. Therefore, a possible fermentation of TMR induced by micro-organism is generally accepted. The present study hypothesized that different lactic acid bacteria could alter fermentation of TMR and subsequently rumen fermentation. Three lactic acid bacteria, Lactobacillus paracasei (A), L. plantarum (B) and L. parabuchneri (C), were employed and 7 treatments under full factorial design were compared with control without inoculation. TMR for dairy cow was used. Significant alterations by treatments were detected at lactic acid and butyric acid contents in TMR (p<0.05). Treatment AC (mixture of A and C) and BC (mixture of B and C) showed great lactate production. Great butyrate production was found at treatment C. At in vitro rumen fermentation, treatments B, C and AB (mixture of A and B) showed significantly great total gas production (p<0.05). All treatments except treatments B and AB, showed less dry matter digestibility, significantly (p<0.05). Total volatile fatty acid production at treatment AC was significantly greater than others (p<0.05). In individual volatile fatty acid production, treatment AB and AC showed great acetate and propionate productions, significantly (p<0.05). This study investigated correlation between organic acid production in TMR and rumen volatile fatty acid production. And it was found that butyric acid in TMR had significant negative correlation with acetate, propionate, total volatile fatty acid, AP ratio and dry matter digestibility.

Preparation of an Anion Exchange Membrane Using the Blending Polymer of Poly(ether sulfone) (PES) and Poly(phenylene sulfide sulfone) (PPSS) (폴리에테르설폰-폴리페닐렌설파이드설폰 블렌딩 고분자를 이용한 음이온교환막의 제조)

  • Lee, Kyung-Han;Han, Joo-Young;Ryu, Cheol-Hwi;Hwang, Gab-Jin
    • Membrane Journal
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    • v.29 no.3
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    • pp.155-163
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    • 2019
  • The anion exchange membrane using the blending polymer of poly(ether sulfone) and poly(phenylene sulfide sulfone) was prepared. It was confirmed by EDXS and FT-IR analysis that the prepared anion exchange membrane had the -N- as an anion exchange group. The ionic conductivity in 1 mol/L $H_2SO_4$ aqueous solution was measured. The ionic conductivity of the prepared anion exchange membrane was 0.015~0.083 S/cm, and had a high value compared with AFN and APS as a commercial anion exchange membrane. Permeabilities of the vanadium ions through the prepared anion exchange membrane were tested to evaluate the possibility as a separator in vanadium redox flow battery. Vanadium ion permeation rate in the prepared anion exchange membrane had a low value compared with Nafion 117 as a commercial cation exchange and AFN as a commercial anion exchange membrane.

Characterization of Antimicrobial Substance Produced by Lactobacillus sp. HN 235 Isolated from Pig Intestine (돼지 장관으로부터 분리한 Lactobacillus sp. HN 235 균주가 생산하는 항균물질의 특성)

  • Shin, Myeong-Su;Han, Sun-Kyung;Choi, Ji-Hyun;Ji, Ae-Ran;Kim, Kyeong-Su;Lee, Wan-Kyu
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.125-132
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    • 2009
  • In order to develop probiotics which may be a viable alternative of antibiotic use in pig industry, five bacterial strains (Lactobacillus sp. HN 52, 92, 98, 235 and AP 116) possessing antimicrobial properties were selected from 500 strains isolates of pig intestines. The bacteriocin produced by Lactobacillus sp. HN 235 displayed a relative broad spectrum of inhibitory activity against all Enterococcus strains, Pseudomonas aeruginosa, Listeria monocytogenes and Clostridium perfringens using the spot-on-lawn method. The production of antimicrobial substance started in the middle of the exponential growth phase, reached maximum levels (6,400 AU/mL) in the stationary phase, and then declined. Bacteriocin activity remained unchanged after 30 min of heat treatment at $95^{\circ}C$ and stable from pH 2.0 to 10 for 1 h, or exposure to organic solvents; however, it diminished after treatment with proteolytic enzymes. The molecular weight of the bacteriocin was about 5 kDa according to a tricine SDS-PAGE analysis.

Anti-wrinkle Activity of Low Molecular Weight Peptides Derived from the Collagen Isolated from Asterias amurensis (불가사리(Asterias amurensis) 콜라겐 유래 저분자 펩타이드의 피부주름 억제활성)

  • Kwon, Min-Chul;Kim, Cheol-Hee;Kim, Hyo-Sung;Syed, Abdul Qadir;Hwang, Bo-Yong;Lee, Hyeon-Yong
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.625-629
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    • 2007
  • This study was carried out to investigate the anti-wrinkle effects of peptides derived from collagens isolated from Asterias amurensis, which was collected in the East Sea. The molecular weights of the peptides were between 10-50 kDa, as determined through sephadek G-75 gel. The cytotoxicities against CCD-986sk cells and HEL-299 cells were measured using the MTT assay. The cytotoxicity of all the fractions(F1: Fraction No. 4-13, 116 kDa; F2: Fraction No. 25-30, 100 kDa; F3: Fraction No. 45-55, 58 kDa; F4: Fraction No. 59-63, 43 kDa; F5: Fraction No. 79-90, 24 kDa) was less than 25%, by the addition of 1.0 mg/mL. These peptides did not show any adverse effects on human skin cells. In the presence of F1 at 1.0 mg/mL, matrix metalloproteinase-1 (MMP-1) expression of UVA-induced human normal fibroblasts was reduced to 34.8%. Overall, the results seem to suggest that peptides of approximately 20 kDa have superior anti-wrinkle effects.

Comparative Effects of Phytase Derived from Escherichia coli and Aspergillus niger in Sixty Eight-week-old Laying Hens Fed Corn-soy Diet

  • Yan, L.;Zhou, T.X.;Jang, H.D.;Hyun, Y.;Kim, H.S.;Kim, I.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.10
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    • pp.1391-1399
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    • 2009
  • Two hundred and sixteen 68-week-old Hy-Line brown laying hens were used in a 6-week feeding trial to compare the efficacy of phytases Optiphos (OPT) and Natuphos (NAT), which were isolated from Escherichia coli and Aspergillus niger, respectively. Hens were randomly allotted into six treatments with six replications (six layers in three adjacent cages) per treatment according to their initial BW. The hens were then subjected to one of the following dietary treatments: i) Positive Control (PC; available phosphorus (AP) 0.4%); ii) Negative control (NC; AP 0.2%); iii) NAT1 (NC+250 FTU/kg NAT); iv) NAT2 (NC+500 FTU/kg NAT); v) OPT1 (NC+250 FTU/kg OPT); vi) OPT2 (NC+500 FTU/kg OPT). Feed intake, egg production, egg quality, apparent nutrient digestibility and serum P and Ca concentration were evaluated to compare the effect of the two phytases. Feed intake and eggshell thickness were not affected by the treatments. Superior effects (p<0.05) of OPT were only observed in egg production and egg weight compared with NAT. Characteristics such as eggshell breaking strength, apparent digestibility of N, Ca and P and serum P concentration were equally increased with the supplementation of both phytases (p<0.05), where no significant difference was observed in those characteristics between PC and phytase supplementation at 500 FTU/kg. Equally effective improvements (p<0.05) were also observed in egg production and DM digestibility, where no improvements were observed (p<0.05) between the PC group and the groups with phytase supplementation at 500 FTU/kg. Equal increases in the serum Ca level were observed when the groups with phytase supplementation were compared to the PC group. Overall, the results of this study suggest that NAT and OPT are equally effective at liberating phytate-bound complexes when included in 0.2% available phosphorus diets for 68-week laying hens; either source of phytase can be fed to commercial 68-week laying hens at 500 FTU/kg to correct the negative effects associated with a 0.2% available phosphorus diet. In conclusion, either source of phytase can be fed to commercial first cycle laying hens at 500 FTU/kg to effectively replace inorganic phosphorus when economically justified.

Sequence and Expression Analysis of c-fos Proto-oncogene in Korean Cattle (HANWOO) (한우 c-fos 유전자의 염기서열 및 발현분석)

  • Yu, S.L.;Chung, H.J.;Jung, K.C.;Lee, J.H.;Cho, K.W.;Choi, J.G.;Na, K.J.;Sang, B.C.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.891-900
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    • 2003
  • Cellular FOS(c-fos) protein is a transcription factor that forms heterodimers mostly with c-jun family and stimulates the transcription of genes containing AP-1 regulatory elements. This c-fos expression can control growth and differentiation of various precursor cells including myoblasts. The controls by c-fos gene have been identified for affecting skeletal muscle fiber traits which are the key determinants of meat quality in pigs. As a first step for identifying the relationship between c-fos gene and meat quality traits in cattle, we fully sequenced 1,443 bp of Hanwoo c-fos mRNA and analyzed expression patterns from various organs and muscle tissues. The sequence identities of Hanwoo c-fos with that of human, pig and mouse showed 89.8%, 93.3% and 87%, respectively. Analyses of the northern blot showed high c-fos expressions were obtained in spleen and rib muscle from 7 organs and 9 different parts of muscles investigated. These results presented here can be used as a valuable marker for meat quality related traits in cattle with further verification.

Study on the Estrogen Receptor Mediated Toxicity of Cadmium and Protective Effects of Antioxidant (에스트로겐 수용체를 통한 카드뮴 독성 및 항산화제에 의한 독성경감에 관한 연구)

  • Kim Tae-Sung;Kang Tae-Seok;Kang Ho-Il;Moon Hyun-Ju;Kang Il-Hyun;Lee Young-Joo;Choi Eun-Hee;Hong Jin-Tae;Han Soon-Young;Hong Jin-Hwan
    • Environmental Mutagens and Carcinogens
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    • v.26 no.1
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    • pp.12-19
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    • 2006
  • Cadmium, a human carcinogen, can induce toxicity in various cell lines and organs. Despite extensive research, the mechanisms of cadmium-induced cell toxicity and estrogenic potential in human are not clear. This study was performed to investigate cadmium-induced toxicity on human breast cancer cells: MCF-7 cells, an estrogen receptor (ER) positive breast cancer cells, and MDA-MB-231 cells, an ER negative breast cancer cells. MCF-7 cells was proved to be more sensitive than the other cell lines (IC50 = $50\;{\mu}M$ at MCF-7 cells and $120{\mu}M$ at MDA-MB-231). The expression of JNK and AP-1 transcription factors such as c-Jun and c-Fos dependent transcription were increased by cadmium treatment. Inhibition of ER activation by ER antagonist (tamoxifen or ICI 182,780) significantly recovered the viablity and inhibited apoptotic cell death. This suggested that cadmium-induced cell death in ER (+) cells was mediated by JNK/AP-1 pathway and this pathway was more stimulated by ER activated by cadmium. Co-treatment of antioxidants such as selenium (Se), butylated hydroxyanisole (BHA), glutathione (GSH), or N-acetyl-L-cysteine (NAC) recovered the cadmium-induced cell death in MCF-7 cells. Cadmium-induced lipid peroxidation was decreased by GSH, NAC, or BHA in MCF-7 cells. The expression of SOD protein was decreased by cadmium ($100{\mu}M$) but recovered by GSH, NAC, BHA, or Se. Our data showed that the cadmium-induced cell toxicity in human breast cancer cells could be protected by the antioxidants (Se, BHA, NAC, GSH, or NAC) and ER antagonist (tamoxifen or ICI 182,780). Therefore, toxicity of cadmium in breast cancer were mediated by oxidative stress and $ER{\alpha}$.

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High $K^+$-Induced Relaxation by Nitric Oxide in Human Gastric Fundus

  • Kim, Dae-Hoon;Kim, Young-Chul;Choi, Woong;Yun, Hyo-Young;Sung, Ro-Hyun;Kim, Hun-Sik;Kim, Heon;Yoo, Ra-Young;Park, Seon-Mee;Yun, Sei-Jin;Song, Young-Jin;Xu, Wen-Xie;Lee, Sang-Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.5
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    • pp.297-303
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    • 2012
  • This study was designed to elucidate high $K^+$-induced relaxation in the human gastric fundus. Circular smooth muscle from the human gastric fundus greater curvature showed stretch-dependent high $K^+$ (50 mM)-induced contractions. However, longitudinal smooth muscle produced stretch-dependent high $K^+$-induced relaxation. We investigated several relaxation mechanisms to understand the reason for the discrepancy. Protein kinase inhibitors such as KT 5823 (1 ${\mu}M$) and KT 5720 (1 ${\mu}M$) which block protein kinases (PKG and PKA) had no effect on high $K^+$-induced relaxation. $K^+$ channel blockers except 4-aminopyridine (4-AP), a voltage-dependent $K^+$ channel ($K_V$) blocker, did not affect high $K^+$ -induced relaxation. However, N(G)-nitro-L-arginine and 1H-(1,2,4)oxadiazolo (4,3-A)quinoxalin-1-one, an inhibitors of soluble guanylate cyclase (sGC) and 4-AP inhibited relaxation and reversed relaxation to contraction. High $K^+$-induced relaxation of the human gastric fundus was observed only in the longitudinal muscles from the greater curvature. These data suggest that the longitudinal muscle of the human gastric fundus greater curvature produced high $K^+$-induced relaxation that was activated by the nitric oxide/sGC pathway through a $K_V$ channel-dependent mechanism.

Difference of Neuronal Recovery by Incubation Condition after Transient Hypoxia (배양조건에 의한 일과성 저산소상태 후 신경세포회복의 차이)

  • Moon, Soo-Hyeon;Oh, Jae-Inn;Park, Youn-Kwan;Chung, Heung-Sub;Lee, Hoon-Kap;Lee, Ki-Chan
    • Journal of Korean Neurosurgical Society
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    • v.29 no.9
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    • pp.1161-1170
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    • 2000
  • Objective : The transverse hippocampal slice is one of the most commonly studied in vitro models of mammalian brain physiology. However, despite its broad usage, there has been no standardization of slice preparation techniques or recording condition. It is well known that variations in recording conditions can result in profound different effects to neuronal responses. Evoked field potentials, recorded extracellularly, were used to investigate the effects of variations in hippocampal slice preparation protocol on hypoxia responses of CA1 neurones. Material & Methods : Before hypoxic injury, hippocampal slices were incubated for 4 hours. During incubation period, the slices were placed in a incubation chamber($21^{\circ}C$) for recovery from preparation injury and then transferred to recording chamber($34^{\circ}C$) for more recovery and baseline electric recording with current stimulation(0.1Hz). Various time periods in incubation chamber and recording chamber were applied to each experimental group(group 1=60min : 180min, group 2=90min : 150min, group 3=180min : 60min, time in incubation chamber : time in recording chamber) before 10 min hypoxia produced by replacing 95% $O_2$+5% $CO_2$ mixed gas to 95% $N_2$+5% $CO_2$ gas. Calcium, Magnesium ions and several drugs effecting on glutamate receptor also were studied. Recoveries from hypoxic injury of hippocampal slices were estimated by percent recovery of population spike(PS). Statistic analysis of study were performed using paired t-test. Results : The percent recovery of PS after 10min hypoxia was considerably enhanced by increasing the period of current stimulation during incubation period before hypoxic injury. Temperature effect on the result of this experiment was also studied(group 4) but the result from this showed no statistic significance. Low magnesium ion concentration of artificial CSF(Mg-free aCSF) during incubation period enhanced the recovery of PS but low calcium (calcium-free) and high magnesium ion concentration(2mM) reduced it after hypoxic injury. L-glutamate($100{\mu}M$) and AP-5($50{\mu}M$) had no effect on the recovery of PS but CNQX($10{\mu}M$) in artificial CSF during incubation period markedly enhanced the recovery of PS. Co-treatment of AP-5($50{\mu}M$), CNQX($10{\mu}M$) and high magnesium concentration(2mM) enhanced recovery of PS in immediate following period of hypoxic injury but the effect of cotreatment after then decayed rapidly and lost statistic significance. Conclusions : Judging from above results, the condition of baseline recording is important in observing the recovery of population spike after hypoxia, and the time and the condition should be controled more strictly to obtain reliable results.

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