• Title/Summary/Keyword: constitutive expression

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Effects of Semen jugrandis on the iNOS Expression and Superoxide Formation in the RAW264.7 Cells (호도(胡挑) 추출물이 마우스 대식세포주인 RAW264.7 세포주의 iNOS 발현 및 Superoxide 형성에 미치는 영향)

  • Moon, Goo;Ko, Su-Mi
    • The Journal of Korean Medicine
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    • v.20 no.1 s.37
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    • pp.151-160
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    • 1999
  • Nitric oxide(NO) is synthesized via the oxidation of L-arginine by a family of nitric oxide synthases(NOS), which are either constitutive(cNOS) or inducible(iNOS). The induction of iNOS in tissues can lead to the sustained production of high concentrations of NO which may exert pro-inflammatory effects including vasodilation. edema, cyototoxicity, and its activity can be mediated by various pro-inflammatory cytokine, including interferon ${\gamma}(INF-{\gamma})$. tumor necrosis factor, IL- 1 and IL-6. The enzyme, iNOS, became a new target for pharmacologcal research with the aim to find new substances for the treatment of chronic inflammatory disorders. Murine macrophages produce large amounts of NO when activated with $TFN-{\gamma}$ plus LPS. The murine macrophage-like cell line, RAW 264.7, is a suitable cell model on which to perform vitro studies regarding the iNOS system. Semen jugrandis is a fatty walnut seed found in Korea. The walnut have been used in foik medicine to improve virility, to relieved asthma, and to relieve constipation. Sesquiterpenelactones were isolated from this plant. In the course of screening for NO inhibitory activity from medicnial plants, the aqueous extract of this plant was found to have a significant activity. The result are summarized as followings. 1. The viability of cells incubated in the presence of semen jugrandis increased mare than non incubated cells. 2. Semen jugrandis suppressed the production of NO in tissues dependent on density. 3. Semen jugrandis suppressed the induction of iNOS in tissues dependent on density can lead to reduced production of NO. 4. Semen jugrandis suppressed the production of superoxide in tissue depend on density. According to the above mentioned results, semen jugrandis could be applied production of NO and superoxide can lead to reduction of chronic inflammatary. And as a depence matter come into a virus of microbe and tumor cells.

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Nonlinear large deflection buckling analysis of compression rod with different moduli

  • Yao, Wenjuan;Ma, Jianwei;Gao, Jinling;Qiu, Yuanzhong
    • Structural Engineering and Mechanics
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    • v.54 no.5
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    • pp.855-875
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    • 2015
  • Many novel materials exhibit a property of different elastic moduli in tension and compression. One such material is graphene, a wonder material, which has the highest strength yet measured. Investigations on buckling problems for structures with different moduli are scarce. To address this new problem, firstly, the nondimensional expression of the relation between offset of neutral axis and deflection curve is derived based on the phased integration method, and then using the energy method, load-deflection relation of the rod is determined; Secondly, based on the improved constitutive model for different moduli, large deformation finite element formulations are developed and combined with the arc-length method, finite element iterative program for rods with different moduli is established to obtain buckling critical loads; Thirdly, material mechanical properties tests of graphite, which is the raw material of graphene, are performed to measure the tensile and compressive elastic moduli, moreover, buckling tests are also conducted to investigate the buckling behavior of this kind of graphite rod. By comparing the calculation results of the energy method and finite element method with those of laboratory tests, the analytical model and finite element numerical model are demonstrated to be accurate and reliable. The results show that it may lead to unsafe results if the classic theory was still adopted to determine the buckling loads of those rods composed of a material having different moduli. The proposed models could provide a novel approach for further investigation of non-linear mechanical behavior for other structures with different moduli.

Transformation of Orchardgrass (Dactylis glomerata L.) with Heat Shock Protein Gene (Heat Shock Protein 유전자를 이용만 오차드그래스의 형질전환)

  • 이효신;이인애;김미혜;손대영;정민섭;조진기
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.75-79
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    • 2001
  • An experiment was carried out to introduce OsHSP17.9, a low molecular HSP gene isolated from rice plant to orchardgrass (Dactylis glomerata L.) using Agrobacterium. Mature seed-derived calli of orchardgrass were co-cultured with Agrobacterium tumefaciens EHA101 harboring the plasmid pIG-HSP17.9 for transformation. Calli selected by hygromycin were transferred to N$_{6}$ medium containing 1 mg/L NAA, 5 mg/L kinetin, 250 mg/L cefotaxime and 50 mg/L hygromycin and several hygromycin resistant plants were obtained. Stable incorporation of the introduced OsHSP17.9 to the genome of the hygromycin resistant plants was confirmed by PCR and Southern blot analysis. Transformation efficiency was variable between cultivars in which it was 16.5% in Potomac and 8.0% in Frontier. Constitutive expression of the transgene in the transformed orchardgrass tissues was identified by Northern blot analysis but transcript levels were different among individual plants.s.

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PKC Downstream of PI3-Kinase Regulates Peroxynitrite Formation for Nrf2-Mediated GSTA2 Induction

  • Kim, Sang-Geon;Kim, Sun-Ok
    • Archives of Pharmacal Research
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    • v.27 no.7
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    • pp.757-762
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    • 2004
  • The protective adaptive response to electrophiles and reactive oxygen species is mediated by the induction of phase II detoxifying genes including glutathione S-transferases (GSTs). NF-E2-related factor-2 (Nrf2) phosphorylation by protein kinase C (PKC) is a critical event for its nuclear translocation in response to oxidative stress. Previously, we have shown that peroxynitrite plays a role in activation of Nrf2 and Nrf2 binding to the antioxidant response element (ARE) via the pathway of phosphatidylinositol 3-kinase (PI3-kinase) and that nitric oxide synthase in hepatocytes is required for GSTA2 induction. In view of the importance of PKC and Pl3-kinase in Nrf2-mediated GST induction, we investigated the role of these kinases in peroxynitrite formation for GSTA2 induction by oxidative stress and determined the relationship between PKC and PI3-kinase. Although PKC activation by phorbol 12-myristate-13-acetate (PMA) did not increase the extents of constitutive and inducible GSTA2 expression, either PKC depletion by PMA or PKC inhibition by staurosporine significantly inhibited GSTA2 induction by tert-butylhydroquinone (t-SHa) a prooxidant chemical. Therefore, the basal PKC activity is req- uisite for GSTA2 induction. 3-Morpholinosydnonimine (SIN-1), which decomposes and yields peroxynitrite, induced GSTA2, which was not inhibited by PKC depletion, but slightly enhanced by PKC activation, suggesting that PKC promotes peroxynitrite formation for Nrf2-mediated GSTA2 induction. Treatment of cells with S-nitroso-N-acetyl-penicillamine (SNAP), an exogenous NO donor, in combination with t-BHQ may produce peroxynitrite. GSTA2 induction by SNAP + t-BHQ was not decreased by PKC depletion, but rather enhanced by PKC activation, showing that the activity of PKC might be required for peroxynitrite formation. LY294002 a P13-kinase inhibitor blocked GSTA2 induction by t-BHQ, which was reversed by PMA-induced PKC activation. These results provide evidence that PKC may playa role in formation of peroxynitrite that activates Nrf2 for GSTA2 induction and that PKC may serve an activator for GSTA2 induction downstream of PI3-kinase.

The role of calmodulin in regulating calcium-permeable PKD2L1 channel activity

  • Park, Eunice Yon June;Baik, Julia Young;Kwak, Misun;So, Insuk
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.3
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    • pp.219-227
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    • 2019
  • Polycystic kidney disease 2-like-1 (PKD2L1), polycystin-L or transient receptor potential polycystin 3 (TRPP3) is a TRP superfamily member. It is a calcium-permeable non-selective cation channel that regulates intracellular calcium concentration and thereby calcium signaling. Although the calmodulin (CaM) inhibitor, calmidazolium, is an activator of the PKD2L1 channel, the activating mechanism remains unclear. The purpose of this study is to clarify whether CaM takes part in the regulation of the PKD2L1 channel, and if so, how. With patch clamp techniques, we observed the current amplitudes of PKD2L1 significantly reduced when co-expressed with CaM and $CaM{\triangle}N$. This result suggests that the N-lobe of CaM carries a more crucial role in regulating PKD2L1 and guides us into our next question on the different functions of two lobes of CaM. We also identified the predicted CaM binding site, and generated deletion and truncation mutants. The mutants showed significant reduction in currents losing PKD2L1 current-voltage curve, suggesting that the C-terminal region from 590 to 600 is crucial for maintaining the functionality of the PKD2L1 channel. With PKD2L1608Stop mutant showing increased current amplitudes, we further examined the functional importance of EF-hand domain. Along with co-expression of CaM, ${\triangle}EF$-hand mutant also showed significant changes in current amplitudes and potentiation time. Our findings suggest that there is a constitutive inhibition of EF-hand and binding of CaM C-lobe on the channel in low calcium concentration. At higher calcium concentration, calcium ions occupy the N-lobe as well as the EF-hand domain, allowing the two to compete to bind to the channel.

Wnt/β-Catenin Signaling Pathway Is Necessary for the Specification but Not the Maintenance of the Mouse Retinal Pigment Epithelium

  • Jong-Myeong Kim;Kwang Wook Min;You-Joung Kim;Ron Smits;Konrad Basler;Jin Woo Kim
    • Molecules and Cells
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    • v.46 no.7
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    • pp.441-450
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    • 2023
  • β-Catenin (Ctnnb1) has been shown to play critical roles in the development and maintenance of epithelial cells, including the retinal pigment epithelium (RPE). Ctnnb1 is not only a component of intercellular junctions in the epithelium, it also functions as a transcriptional regulator in the Wnt signaling pathway. To identify which of its functional modalities is critically involved in mouse RPE development and maintenance, we varied Ctnnb1 gene content and activity in mouse RPE lineage cells and tested their impacts on mouse eye development. We found that a Ctnnb1 double mutant (Ctnnb1dm), which exhibits impaired transcriptional activity, could not replace Ctnnb1 in the RPE, whereas Ctnnb1Y654E, which has reduced affinity for the junctions, could do so. Expression of the constitutively active Ctnnb1∆ex3 mutant also suppressed the development of RPE, instead facilitating a ciliary cell fate. However, the post-mitotic or mature RPE was insensitive to the loss, inactivation, or constitutive activation of Ctnnb1. Collectively, our results suggest that Ctnnb1 should be maintained within an optimal range to specify RPE through transcriptional regulation of Wnt target genes in the optic neuroepithelium.

Effects of freeze-thaw cycle on mechanical properties of saline soil and Duncan-Chang model

  • Shukai Cheng;Qing Wang;Jiaqi Wang;Yan Han
    • Geomechanics and Engineering
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    • v.38 no.3
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    • pp.249-260
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    • 2024
  • In order to study the mechanical propertied and change rules of undrained shear behavior of saline soil under the freeze-thaw cycles, an improved constitutive model reflecting the effects of freeze-thaw cycles was proposed based on the traditional Duncan-Chang model. The saline soil in Qian'an County, western Jilin Province, was selected as the experimental object. Then, a set of freeze-thaw cycles (0, 1, 10, 30, 60, 90, 120) tests were conducted on the saline soil specimens, and conventional consolidated undrained triaxial shear tests were conducted on the saline soil specimens that underwent freeze-thaw cycles. The stress-strain relationship was obtained by the triaxial shear test. The model parameters have a corresponding regression relationship with the number of freeze-thaw cycles. Finally, based on the function expression of the model parameters, the modified Duncan-Chang model with the number of freeze-thaw cycles as the influence factor was established, whilst the calculation program of the modified model is compiled. Based on the test results, the stress-strain relationship of the saline soil specimen shows strain hardening. The shear strength gradually decreases with the increase of freeze-thaw cycle. The 10 freeze-thaw cycles are the turning point in the trend of changes of the mechanical properties of saline soils. The calculated and experimental stress-strain relationship are compared, and the comparison between the calculated value of the model and the experimental value showed that the two had a good consistency, which verified the validity of the modified Duncan-Chang model in reflecting the effects of the freeze-thaw cycle.

Identification and characterization of the MYC2 gene in relation to leaf senescence response in hybrid poplar (Populus alba × P. glandulosa) (현사시나무에서 MYC2 유전자의 분리 및 노화 지연에 관한 특성 구명)

  • Choi, Hyunmo;Bae, Eun-Kyung;Cho, Jin Seong;Lee, Hyoshin;Choi, Young-Im
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.409-415
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    • 2017
  • The vegetation period of trees might be prolonged by the delay of the leaf senescence in autumn. Thus, we focused on the generation of senescence-delayed transgenic trees to enhance biomass production. The PagMYC2, a gene containing the basic helix-loop-helix domain, was selected as a candidate for a senescence-delayed transgenic tree. The PagMYC2 gene was specifically induced after treatment with phytohormone jasmonic acid, and upregulated by abiotic stresses such as salinity, osmotic pressure and a low temperature. The constitutive overexpression of the PagMYC2 delayed the leaf senescence and inhibited chlorophyll degradation in the transgenic poplars. Leaf senescence analysis was performed in the leaf tissues of the PagMYC2-over-expression transgenic poplars. The transgenic poplars exhibited higher photochemical efficiency than did a wild type plant under a short-day condition (6 hours light/18 hours darkness) or a low temperature condition ($15^{\circ}C$) that was similar to the weather conditions of autumn. These results suggest that the PagMYC2 is a useful genetic resource to improve biomass production, which is able to sustain growth with senescence-delayed leaves for a long time in autumn.

Identification of disease resistance to soft rot in transgenic potato plants that overexpress the soybean calmodulin-4 gene (GmCaM-4) (대두 칼모듈린 단백질, GmCaM-4를 발현하는 형질전환 감자의 무름병 저항성 확인)

  • Park, Hyeong Cheol;Chun, Hyun Jin;Kim, Min Chul;Lee, Sin Woo;Chung, Woo Sik
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.157-163
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    • 2020
  • Calmodulin (CaM) mediates cellular Ca2+ signals in the defense responses of plants. We previously reported that GmCaM-4 and 5 are involved in salicylic acid-independent activation of disease resistance responses in soybean (Glycine max). Here, we generated a GmCaM-4 cDNA construct under the control of the cauliflower mosaic virus (CaMV) 35S promoter and transformed this construct into potato (Solanum tuberosum L.). The constitutive over-expression of GmCaM-4 in potato induced high-level expression of pathogenesis-related (PR) genes, such as PR-2, PR-3, PR-5, phenylalanine ammonia-lyase (PAL), and proteinase inhibitorII (pinII). In addition, the transgenic potato plants exhibited enhanced resistance against a bacterial pathogen, Erwinia carotovora ssp. Carotovora (ECC), that causes soft rot disease and showed spontaneous lesion phenotypes on their leaves. These results strongly suggest that a CaM protein in soybean, GmCaM-4, plays an important role in the response of potato plants to pathogen defense signaling.

Methylated Alteration of SHP1 Complements Mutation of JAK2 Tyrosine Kinase in Patients with Myeloproliferative Neoplasm

  • Yang, Jun-Jun;Chen, Hui;Zheng, Xiao-Qun;Li, Hai-Ying;Wu, Jian-Bo;Tang, Li-Yuan;Gao, Shen-Meng
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2219-2225
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    • 2015
  • SHP1 negatively regulates the Janus kinase 2/signal transducer and activator of transcription (JAK2/STAT) signaling pathway, which is constitutively activated in myeloproliferative neoplasms (MPNs) and leukemia. Promoter hypermethylation resulting in epigenetic inactivation of SHP1 has been reported in myelomas, leukemias and other cancers. However, whether SHP1 hypermethylation occurs in MPNs, especially in Chinese patients, has remained unclear. Here, we report that aberrant hypermethylation of SHP1 was observed in several leukemic cell lines and bone marrow mononuclear cells from MPN patients. About 51 of 118 (43.2%) MPN patients including 23 of 50 (46%) polycythaemia vera patients, 20 of 50 (40%) essential thrombocythaemia and 8 of 18 (44.4%) idiopathic myelofibrosis showed hypermethylation by methylation-specific polymerase chain reaction. However, SHP1 methylation was not measured in 20 healthy volunteers. Hypermethylation of SHP1 was found in MPN patients with both positive (34/81, 42%) and negative (17/37, 45.9%) JAK2V617F mutation. The levels of SHP1 mRNA were significantly lower in hypermethylated samples than unmethylated samples, suggesting SHP1 may be epigenetically inactivated in MPN patients. Furthermore, treatment with 5-aza-2'-deoxycytidine (AZA) in K562 cells showing hypermethylation of SHP1 led to progressive demethylation of SHP1, with consequently increased reexpression of SHP1. Meanwhile, phosphorylated JAK2 and STAT3 were progressively reduced. Finally, AZA increased the expression of SHP1 in primary MPN cells with hypermethylation of SHP1. Therefore, our data suggest that epigenetic inactivation of SHP1 contributes to the constitutive activation of JAK2/STAT signaling. Restoration of SHP1 expression by AZA may contribute to clinical treatment for MPN patients.