• Title/Summary/Keyword: loxP

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Modification of Animal Genotypes for the Regulation of Transgene Expression (이식유전자 발현조절을 위한 동물유전자의 조작)

  • 진동일
    • Korean Journal of Animal Reproduction
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    • v.19 no.4
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    • pp.283-291
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    • 1996
  • Transgenic animal을 응용할 수 있는 분야에서는 이식유전자의 기능을 정확하게 규명하고 이를 바탕으로 실질적인 유전적인 개량을 이루기 위해서 이식유전자의 발현을 조절할 수 있는 정교한 system이 필요하다. 유전자의 미세주입법에 의해 transgenic animal을 생산할 수 있는데 이용되고 있는 tissue-specific promoter에 의한 이식유전자의 발현조절은 필요로 하는 시기나 양 등을 인위적으로 조절하고자 하는데 한계점을 갖고 있다. 이러한 이식유전자 발현의 문제점을 극복하기 위해 효모의 recombinase나 미생물의 repressor 단백질과 이들의 binding site인 operator sequence를 이용하여 인위적으로 이식유전자의 발현을 조절할 수 있는 system이 개발되고 있다. Cre/loxP system은 site-specific recombination에 의해 DNA sequence를 제거함으로서 이식유전자의 발현을 조절할 수 있다. 이식유전자 발현의 장소와 양을 조절하기 위해서는 미생물이 이용하고 있는 repressor와 이들의 operator sequence를 적용하여 ligand binary system이 개발되었다. Lac repressor system에서는 isopropyl-$\beta$-D-thiogalactoside (IPTG)가 이식유전자 발현을 조절할 수 있는 positive regulator로서 작용하고, tetracycline-VP16 system에서는 tetracycline이나 유사물질들이 negative regulator로서 이용할 수 있다. 이러한 binary system은 transgenic animal에서 이식유전자 발현의 장소와 시기 또한 양을 효과적으로 조절하는데 적용할 수 있는 것으로 나타났다. 따라서 기존의 binary system과 함께 새로운 regulatory system의 장점을 이용하여 보다 완벽한 이식유전자의 인위적인 조절 system을 이룩함으로서 transgenic animal technology의 실용화를 앞당길 것으로 기대된다.

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Study on the Temperature Characteristic of Pressurization System Using Cryogenic Helium Gas (극저온 헬륨가스 가압시스템에 대한 온도특성 연구(I))

  • Chung Yonggahp;Kim Yoo
    • Journal of the Korean Society of Propulsion Engineers
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    • v.9 no.3
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    • pp.66-73
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    • 2005
  • The pressurization system in a liquid rocket propulsion system provides a controlled gas pressure in the ullage space of the vehicle propellant tanks. It is advantage to employ a hot gas heat exchanger in the pressurization system to increase the specific volume of the pressurant and thereby reduce over-all system weight. A significant improvement in pressurization-system performance can be achieved, particularly in a cryogenic system, where the gas supply is stored inside the cryogenic propellant tank. In this study liquid nitrogen was used instead of liquid oxygen as a simulant. The temperature characteristic of cryogenic pressurant is very important to develop some components in pressurization system. Numerical modeling and test data were studied using SINDA/FLUINT Program and PTF(Propellant-feeding Test facility).

Olig2 Transcription Factor in the Developing and Injured Forebrain; Cell Lineage and Glial Development

  • Ono, Katsuhiko;Takebayashi, Hirohide;Ikenaka, Kazuhiro
    • Molecules and Cells
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    • v.27 no.4
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    • pp.397-401
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    • 2009
  • Olig2 transcription factor is widely expressed throughout the central nervous system; therefore, it is considered to have multiple functions in the developing, mature and injured brain. In this mini-review, we focus on Olig2 in the forebrain (telencephalon and diencephalon) and discuss the functional significance of Olig2 and the differentiation properties of Olig2-expressing progenitors in the development and injured states. Short- and long-term lineage analysis in the developing forebrain elucidated that not all late Olig2+ cells are direct cohorts of early cells and that Olig2 lineage cells differentiate into neurons or glial cells in a region- and stage-dependent manner. Olig2-deficient mice revealed large elimination of oligodendrocyte precursor cells and a decreased number of astrocyte progenitors in the dorsal cortex, whereas no reduction in the number of GABAergic neurons. In addition to Olig2 function in the developing cortex, Olig2 is also reported to be important for glial scar formation after injury. Thus, Olig2 can be essential for glial differentiation during development and after injury.

Antioxidative Effects of Water-Soluble Chitinous Compounds on Oxidation of Low Density Lipoprotein in Macrophages (대식세포에서 지단백 산화에 대한 수용성 Chitinous Compounds의 항산화 효과에 대한 연구)

  • 이세희;박성희;이용진;윤정한;최연정;최정숙;강영희
    • Journal of Nutrition and Health
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    • v.36 no.9
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    • pp.908-917
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    • 2003
  • It has been proposed that oxidative modification of LDL (oxLDL) plays a significant role in the pathogenicity of atherogenesis. We tested the hypothesis that chitin and chitosan may function as antioxidants with respect to 0.1 mg cholesterol/ml LDL incubated with 5 $\mu$ M Cu$^2$$^{+}$alone or in the P338Dl mouse macrophage system using L-ascorbic acid as a standard classical antioxidant. The degree of oxLDL formation was ascertained by the relative electrophoretic mobility (rEM) in the combination of thiobarbituric acid reactive substances (TBARS) levels, and the cytotoxicity of oxLDL was detected by macrophage viability. The oxLDL uptake and foam cell formation of macrophages were measured by Oil Red O staining. Incubation with Cu$^2$$^{+}$and macrophages increased rEM of LDL and stimulated TBARS formation. Culture of macrophages with LDL in the presence 5 $\mu$ M Cu$^2$$^{+}$induced macrophage death. In cell-free system 200 $\mu$g/ml water-soluble chitosan and chitosan-oligosaccharide blocked oxLDL formation. Water-soluble chitosan and chitosan-oligosaccharide blocked oxLDL formation near-completely relative to L-ascorbic acid, whereas water-soluble chitin and chitin-oligosaccharide had no measurable antioxidant effect. In macrophage system water-soluble chitosan and chitosan-oligosaccharide blocked oxidation of LDL with a significant increase in cell viability, and decreased TBARS in medium. As for the inhibitory effect on macrophage foam cell formation, chitosan and its oligosaccharide, but not watersoluble chitin, revealed the effectiveness. The endothelial expression of lectin-like oxLDL receptor-1 (LOX-1) was tested by Western blot analysis, and chitosan, chitosan-oligosaccharide and chitin-oligosaccharide blocked LOX-1 expression. These results indicate that water-soluble chitosan and its oligosaccharide showed the inhibitory effect on Cu$^2$$^{+}$-induced LDL oxidation of macrophages, and chitosan, chitosan-oligosaccharide and chitin-oligosaccharide had blocking effect on oxLDL receptor expression in the human umbilical vein endothelial system. Thus, water-soluble chitosan and its oligosaccharides possess anti-atherogenic potentials possibly through the inhibition of macrophage LDL oxidation or endothelial oxLDL receptor expression depending on chemical types.l types.

Current Progress in Generation of Genetically Modified Mice (유전자 조작 마우스 개발의 최신 연구 동향)

  • Song, Ki-Duk;Cho, Byung-Wook
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.587-592
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    • 2007
  • Manipulation of the mouse genome by activating or inactivating the gene has contributed to the understanding of the function of the gene in the subset of cells during embryonic development or postnatal period of life. Most of all, gene targeting, which largely depends on the availability of mouse embryonic stem (ES) cells, is the milestone of development of animal models for human disease. Recombinase-mediated genome modification (Cre-LoxP and Flp-Frt etc) and the ligand-dependent regulation system, more accurate and elaborate manipulation tools, have been successfully developed and applied to dissect the mechanisms governing complex biological processes and to understand the role of protein in temporal-and spatial aspects of development. As technologies concerning refined manipulation of mouse genome are developed, they are expected to open new opportunities to better understand the diverse in vivo functions of genes.

Arachidonic Acid Activates $K^+$-$Cl^-$-cotransport in HepG2 Human Hepatoblastoma Cells

  • Lee, Yong-Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.5
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    • pp.401-408
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    • 2009
  • $K^+$-$Cl^-$-cotransport (KCC) has been reported to have various cellular functions, including proliferation and apoptosis of human cancer cells. However, the signal transduction pathways that control the activity of KCC are currently not well understood. In this study we investigated the possible role of phospholipase $A_2$ ($PLA_2$)-arachidonic acid (AA) signal in the regulatory mechanism of KCC activity. Exogenous application of AA significantly induced $K^+$ efflux in a dose-dependent manner, which was completely blocked by R-(+)-[2-n-butyl-6,7 -dichloro-2-cyclopentyl-2,3-dihydro-1-oxo-1Hinden-5-yl]oxy]acetic acid (DIOA), a specific KCC inhibitor. N-Ethylmaleimide (NEM), a KCC activatorinduced $K^+$ efflux was significantly suppressed by bromoenol lactone (BEL), an inhibitor of the calciumindependent $PLA_2$ ($iPLA_2$), whereas it was not significantly altered by arachidonyl trifluoromethylketone ($AACOCF_3$) and p-bromophenacyl bromide (BPB), inhibitors of the calcium-dependent cytosolic $PLA_2$ ($cPLA_2$) and the secretory $PLA_2$ ($sPLA_2$), respectively. NEM increased AA liberation in a doseand time-dependent manner, which was markedly prevented only by BEL. In addition, the NEM-induced ROS generation was significantly reduced by DPI and BEL, whereas $AACOCF_3$ and BPB did not have an influence. The NEM-induced KCC activation and ROS production was not significantly affected by treatment with indomethacin (Indo) and nordihydroguaiaretic acid (NDGA), selective inhibitors of cyclooxygenase (COX) and lipoxygenase (LOX), respectively. Treatment with 5,8,11,14-eicosatetraynoic acid (ETYA), a non-metabolizable analogue of AA, markedly produced ROS and activated the KCC. Collectively, these results suggest that $iPLA_2$-AA signal may be essentially involved in the mechanism of ROS-mediated KCC activation in HepG2 cells.

Comparative evaluation of pain perception following topical application of clove oil, betel leaf extract, lignocaine gel, and ice prior to intraoral injection in children aged 6-10 years: a randomized control study

  • Havale, Raghavendra;Rao, Dhanu G;Shrutha, SP;Tuppadmath, Kanchan M;Tharay, Namratha;Mathew, Irin;Taj, Kausar E
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.21 no.4
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    • pp.329-336
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    • 2021
  • Background: In the field of dentistry, topical anesthetics play an important role in reducing pain during needle pricks. The anesthetic property of betel leaves remain unexplored, even though they have been widely used for the treatment of various ailments. The purpose of this study was to compare and evaluate pain perception following topical application of lignocaine gel, clove gel, ice, and newly developed betel leaf extract gel during intraoral injection in children. Methods: Sixty children aged 6-10 years who met the inclusion criteria were divided into four groups. Topical anesthetic agents, 2% lignocaine (Lox-2% Jelly, Neon, Mumbai, India), 4.7% clove gel (Pain Out Dental Gel, Colgate Palmolive India Ltd, Solan, India), 10% betel leaf extract gel, and ice were applied to each group for one min, followed by administration of infiltration anesthesia. Pain perception was analyzed during needle insertion. The Wong Bakers FACES pain rating scale (WBFPRS) was used for subjective assessment and the Sound, Eye, Motor (SEM) scale for objective assessment. Recorded values were tabulated and subjected to appropriate statistical analysis using SPSS software with a P value set at 0.05. Results: The clove oil and betel leaf groups demonstrated the highest WBFPRS scores, followed by the ice and lignocaine groups. The clove, betel leaf extract, and ice groups showed equal and highest SEM scores, followed by the lignocaine group. The mean WBFPRS and mean SEM scores were statistically significant. Conclusion: Betel leaf extract gel is effective in reducing pain and can act as an alternative topical anesthetic agent.

Anti-inflammatory Effects of Goihwa-san Water Extract via NF-κB Inhibition (괴화산(槐花散)의 NF-κB 기전을 통한 항염증 효과 연구)

  • Hyun Hee Cho;Ji Young Choi;Min Hwangbo;Seon Young Jee
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.36 no.1
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    • pp.21-39
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    • 2023
  • Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Goihwa-san water extract(GHS) in vitro & in vivo. Methods : In vitro, we evaluated the anti-inflammatory effect of GHS by comparing the Raw 264.7 cells with 10, 30, 100, 300㎍/㎖ of GHS for 1 hour before Lipopolysaccharide(LPS) to the single LPS treated group. We examined the relative cell viability by MTT assay and the relative level of LPS, Loxoribine(LOX), Peptidoglycan(PGN), Flagellin(FLA)-induced NO production by using Griess reagent and measured relative iNOS protein level and COX-2 protein level by using western blot and Image analyzing system. We measured the production of TNF-α, IL-1β, and IL-6 by each ELISA kits and then measured the relative levels of IκBα, p-IκBα in whole-cell lysate fraction and NF-κB in nuclear fraction by using western blot and Image analyzing system. In vivo, we induced the paw edema by subcutaneous injection of 100㎕/rat CA and measured the swelling volume of paw by using a plethysmometer and then measured the relative iNOS protein level by using western blot. Results : As a result, in vitro, LPS, PGN-induced NO production was significantly inhibited by pretreatment with GHS. GHS reduced LPS, PGN-induced iNOS expression, PGN-induced COX-2 expression and LPS-induced production of cytokine(TNF-α, IL-1β, IL-6). Expression of IκBα was increased by pretreatment with GHS 100㎍/㎖. And the expression of p-IκBα and NF-κB were decreased by pretreatment with GHS 100㎍/㎖. In vivo, CA-induced inflammation rat model was used for the evaluation of the anti-inflammatory effect of GHS. 0.3 or 1.0g/kg of GHS significantly reduced the increases of paw swelling and iNOS expression in paw tissues. Conclusions : These results show that GHS can decrease inflammatory response via inhibition of the NF-κB pathway in vitro. And in vivo, the anti-inflammatory effect suggest the clinical basis of GHS for the treatment of inflammatory diseases.

Inhibitory Effects of Apple Peel Extract on Inflammatory Enzymes (사과 과피 추출물의 염증 관련 효소 억제 효과)

  • Kim, Ilrang
    • Korean Journal of Food Science and Technology
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    • v.47 no.4
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    • pp.534-538
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    • 2015
  • The purpose of this study was to investigate the biological benefits of apple peel. The antioxidant and anti-inflammatory activities of a 70% ethanol extract of apple peel were examined. The total phenolic compound and flavonoid contents of apple peel were $6.8{\pm}0.5mg$ gallic acid equivalent/g of fresh weight and $3.3{\pm}0.3mg$ catechin equivalent/g of fresh weight, respectively. Antioxidant activity was evaluated by measuring 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. The DPPH radical scavenging activity of apple peel was $18.9{\pm}1.6$, $46.3{\pm}2.3$ and $58.1{\pm}3.9%$ at concentrations of 0.1, 0.5 and 1.0 mg/mL, respectively (p<0.05). The anti-inflammatory effect was investigated by measuring the inhibition of inflammatory enzymes. Apple peel significantly inhibited secretory phospholipase, cyclooxygenase-1, cyclooxygenase-2, and lipoxygenase activity by up to $53.5{\pm}2.3$, $13.4{\pm}1.8$, $64.8{\pm}5.4$ and $44.4{\pm}4.5%$, respectively (p<0.05). Taken together, these findings suggest that apple peel may act as an antioxidant by radical scavenging and may possess potential anti-inflammatory properties for suppressing the activity of inflammatory enzymes. These results also suggest that apple peel can be utilized as a health functional food ingredient possessing antioxidant and anti-inflammatory activities.

Hepatoprotective Effects of Oyster Hydrolysate on Lipopolysaccharide/D-Galactosamine-Induced Acute Liver Injury in Mice (Lipopolysaccharide/D-Galactosamine에 의해 유도된 급성 간 손상 생쥐모델에서 굴가수분해물의 간 보호 효과)

  • Ryu, Ji Hyeon;Kim, Eun-Jin;Xie, Chengliang;Nyiramana, Marie Merci;Siregar, Adrian S.;Park, Si-Hyang;Cho, Soo Buem;Song, Dae Hyun;Kim, Nam-Gil;Choi, Yeung Joon;Kang, Sang Soo;Kang, Dawon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.6
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    • pp.659-670
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    • 2017
  • Oxidative stress and inflammation are key factors responsible for progression of liver injury. A variety of functions of oyster hydrolysate (OH) are affected by their antioxidant and anti-inflammatory activities. However, little is known regarding the effects of OH on a liver injury model. This study was performed to evaluate the effects of OH on acute liver injury induced by lipopolysaccharide/D-galactosamine (LPS/D-GalN) in mice. Experimental groups were divided into six groups as follows (each group, n=10): control (saline), LPS/D-GalN, LPS/D-GalN+OH (100 mg/kg), LPS/D-GalN+OH (200 mg/kg), LPS/D-GalN+OH (400 mg/kg), and LPS/D-GalN+silymarin (25 mg/kg, positive control). The experimental acute liver injury model was induced with LPS ($1{\mu}g/kg$) and D-GalN (400 mg/kg). We first analyzed antioxidant and anti-inflammatory activities in OH. OH showed high DPPH and ABTS radical scavenging activities and reduced ROS generation in Chang cells in a dose-dependent manner. In addition, OH showed anti-inflammatory activities, such as inhibition of cyclooxygenase-2 and 5-lipooxygenase. Treatment with OH down-regulated tumor necrosis factor $(TNF)-{\alpha}$, interleukin (IL)-6, and $IL-1{\alpha}$ expression levels in LPS-stimulated RAW264.7 cells. OH significantly reduced LPS/D-GalN-induced increases in the concentrations of alanine transaminase and aspartate aminotransferase in serum. In the LPS/D-GalN group, liver tissues exhibited apoptosis of hepatocytes with hemorrhages. These pathological alterations were ameliorated by OH treatment. Consistently, hepatic catalase activity was low in the LPS/D-GalN group compared to the control group, and catalase activity was significantly restored by OH treatment (P<0.05). Furthermore, OH markedly reduced the LPS/D-GalN-induced increase in $TNF-{\alpha}$, $IL-1{\beta}$, and IL-6 levels in liver tissue. Taken together, these results show that OH has hepatoprotective effects on LPS/D-GalN-induced acute liver injury via inhibition of oxidative stress and inflammation, suggesting that OH could be used as a health functional food and potential therapeutic agent for acute liver injury.