• Title/Summary/Keyword: signaling state

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Effect of all-trans retinoic acid on casein and fatty acid synthesis in MAC-T cells

  • Liao, Xian-Dong;Zhou, Chang-Hai;Zhang, Jing;Shen, Jing-Lin;Wang, Ya-Jing;Jin, Yong-Cheng;Li, Sheng-Li
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.1012-1022
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    • 2020
  • Objective: Caseins and fatty acids of milk are synthesized and secreted by the epithelial cells of the mammary gland. All-trans retinoic acid (ATRA), an active metabolite of vitamin A, has been shown to promote mammary development. This study was conducted to determine the effect of ATRA on casein synthesis and fatty acid composition in MAC-T cells. Methods: MAC-T cells were allowed to differentiate for 4 d, treated with ATRA (0, 1.0, 1.5, and 2.0 μM), and incubated for 3 d. We analyzed the fatty acid composition, the mRNA expression of casein and fatty acid synthesis-related genes, and the phosphorylation of casein synthesis-related proteins of MAC-T cells by gas chromatography, quantitative polymerase chain reaction, and western blotting, respectively. Results: In MAC-T cells, ATRA increased the mRNA levels of αS1-casein and β-casein, janus kinase 2 (JAK2) and E74-like factor 5 of the signal transducer and activator of transcription 5 β (STAT5-β) pathway, ribosomal protein S6 kinase beta-1 (S6K1) and eukaryotic translation initiation factor 4E binding protein 1 of the mammalian target of rapamycin (mTOR) pathway, inhibited the mRNA expression of phosphoinositide 3-kinase and eukaryotic initiation factor 4E of the mTOR pathway, and promoted the phosphorylation of STAT5-β and S6K1 proteins. Additionally, ATRA increased the de novo synthesis of fatty acids, reduced the content of long-chain fatty acids, the ratio of monounsaturated fatty acids to saturated fatty acids (SFA), the ratio of polyunsaturated fatty acids (PUFA) to SFA, and the ratio of ω-6 to ω-3 PUFA. The mRNA levels of acetyl-CoA carboxylase 1, fatty acid synthase, lipoprotein lipase, stearoyl-CoA desaturase, peroxisome proliferator-activated receptor gamma, and sterol regulatory element-binding protein 1 (SREBP1) were enhanced by ATRA. Conclusion: ATRA promotes the synthesis of casein by regulating JAK2/STAT5 pathway and downstream mTOR signaling pathway, and it improves the fatty acid composition of MAC-T cells by regulating SREBP1-related genes.

A Scalable Dynamic QoS Support Protocol (확장성 있는 동적 QoS 지원 프로토콜)

  • 문새롬;이미정
    • Journal of KIISE:Information Networking
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    • v.29 no.6
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    • pp.722-737
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    • 2002
  • As the number of multimedia applications increases, various protocols and architectures have been proposed to provide QoS(Quality of Service) guarantees in the Internet. Most of these techniques, though, bear inherent contradiction between the scalability and the capability of providing QoS guarantees. In this paper, we propose a protocol, named DQSP(Dynamic QoS Support Protocol), which provides the dynamic resource allocation and admission control for QoS guarantees in a scalable way. In DQSP, the core routers only maintain the per source-edge router resource allocation state information. Each of the source-edge routers maintains the usage information for the resources allocated to itself on each of the network links. Based on this information, the source edge routers perform admission control for the incoming flows. For the resource reservation and admission control, DQSP does not incur per flow signaling at the core network, and the amount of state information at the core routers depends on the scale of the topology instead of the number of user flows. Simulation results show that DQSP achieves efficient resource utilization without incurring the number of user flow related scalability problems as with IntServ, which is one of the representative architectures providing end-to-end QoS guarantees.

Rhodopsin Chromophore Formation and Thermal Stabilities in the Opsin Mutant E134Q/M257Y (옵신 mutant E134Q/M257Y의 로돕신 형성과 열안정성 분석)

  • Kim, Jong-Myoung
    • Journal of Life Science
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    • v.22 no.7
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    • pp.863-870
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    • 2012
  • Rhodopsin, a dim light photoreceptor, has been regarded as one of the model systems for the structural and functional study of G protein-coupled receptors (GPCRs). Constitutively active mutant GPCRs leading to the activation of heterotrimeric GDP/GTP-binding protein signaling in the absence of ligand binding are of interest for the study of the activation mechanism in GPCRs. The present study focused on the opsin mutant E134Q/M257Y, which showed a moderate level of constitutive activity and the formation of two distinct rhodopsin chromophores with absorption maxima of 500 nm and 380 nm, depending on the presence of an inverse agonist, 11-cis-retinal, and an agonist, all-trans-retinal, respectively. Reconstitution of the mutant rhodopsin upon incubation with different ratios of 11-cis-retinal and the all-trans-retinal, as well as upon sequential binding of the two retinals, indicated its preferential binding to 11-cis-retinal. The thermal stability of the 11-cis-retinal-bound form of the E134Q/M257Y mutant was lower than that of the mutants containing a single replacement but higher than that of the all-trans-retinal-bound forms. The mutant also showed a lower stability in its opsin state as compared with that of the wild-type opsin but had little effects on the binding affinity to 11-cis-retinal. Information obtained in this study will be helpful for analyzing the structural changes associated with the activation of rhodopsin and GPCRs.

Inactivation of Brain myo-Inositol Monophosphate Phosphatase by Pyridoxal-5'-Phosphate

  • Kim, Dae-Won;Hong, Joung-Woo;Eum, Won-Sik;Choi, Hee-Soon;Choi, Soo-Hyun;Kim, So-Young;Lee, Byung-Ryong;An, Jae-Jin;Lee, Sun-Hwa;Lee, Seung-Ree;Kwon, Oh-Shin;Kwon, Hyeok-Yil;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Choi, Soo-Young
    • BMB Reports
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    • v.38 no.1
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    • pp.58-64
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    • 2005
  • Myo-inositol monophosphate phosphatase (IMPP) is a key enzyme in the phosphoinositide cell-signaling system. This study found that incubating the IMPP from a porcine brain with pyridoxal-5'-phosphate (PLP) resulted in a time-dependent enzymatic inactivation. Spectral evidence showed that the inactivation proceeds via the formation of a Schiff's base with the amino groups of the enzyme. After the sodium borohydride reduction of the inactivated enzyme, it was observed that 1.8 mol phosphopyridoxyl residues per mole of the enzyme dimer were incorporated. The substrate, myo-inositol-1-phosphate, protected the enzyme against inactivation by PLP. After tryptic digestion of the enzyme modified with PLP, a radioactive peptide absorbing at 210 nm was isolated by reverse-phase HPLC. Amino acid sequencing of the peptide identified a portion of the PLP-binding site as being the region containing the sequence L-Q-V-S-Q-Q-E-D-I-T-X, where X indicates that phenylthiohydantoin amino acid could not be assigned. However, the result of amino acid composition of the peptide indicated that the missing residue could be designated as a phosphopyridoxyl lysine. This suggests that the catalytic function of IMPP is modulated by the binding of PLP to a specific lysyl residue at or near its substrate-binding site of the protein.

Regulation of L-type Calcium Channel Current by Somatostatin in Guinea-Pig Gastric Myocytes

  • Kim, Young-Chul;Sim, Jae-Hoon;Lee, Sang-Jin;Kang, Tong-Mook;Kim, Sung-Joon;Kim, Seung-Ryul;Youn, Sei-Jin;Lee, Sang-Jeon;Xu, Wen Xie;So, In-Suk;Kim, Ki-Whan
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.2
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    • pp.103-108
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    • 2005
  • To study the direct effect of somatostatin (SS) on calcium channel current ($I_{Ba}$) in guinea-pig gastric myocytes, $I_{Ba}$ was recorded by using whole-cell patch clamp technique in single smooth muscle cells. Nicardipine ($1{\mu}M$), a L-type $Ca^{2+}$ channel blocker, inhibited $I_{Ba}$ by $98{\pm}1.9$% (n=5), however $I_{Ba}$ was decreased in a reversible manner by application of SS. The peak $I_{Ba}$ at 0 mV were decreased to $95{\pm}1.5$, $92{\pm}1.9$, $82{\pm}4.0$, $66{\pm}5.8$, $10{\pm}2.9$% at $10^{-10}$, $10^{-9}$, $10^{-8}$, $10^{-7}$, $10^{-5}$ M of SS, respectively (n=3∼6; $mean{\pm}SEM$). The steady-state activation and inactivation curves of $I_{Ba}$ as a function of membrane potentials were well fitted by a Boltzmann equation. Voltage of half-activation ($V_{0.5}$) was $-12{\pm}0.5$ mV in control and $-11{\pm}1.9$ mV in SS treated groups (respectively, n=5). The same values of half-inactivation were $-35{\pm}1.4$ mV and $-35{\pm}1.9$ mV (respectively, n=5). There was no significant difference in activation and inactivation kinetics of $I_{Ba}$ by SS. Inhibitory effect of SS on $I_{Ba}$ was significantly reduced by either dialysis of intracellular solution with $GDP_{\beta}S$, a non-hydrolysable G protein inhibitor, or pretreatment with pertussis toxin (PTX). SS also decreased contraction of guinea-pig gastric antral smooth muscle. In conclusion, SS decreases voltage-dependent L-type calcium channel current ($VDCC_L$) via PTXsensitive signaling pathways in guinea-pig antral circular myocytes.

Down-regulation of Phosphoglucose Isomerase/Autocrine Motility Factor Enhances Gensenoside Rh2 Pharmacological Action on Leukemia KG1α Cells

  • You, Zhi-Mei;Zhao, Liang;Xia, Jing;Wei, Qiang;Liu, Yu-Min;Liu, Xiao-Yan;Chen, Di-Long;Li, Jing
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1099-1104
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    • 2014
  • Aims and Background: Ginsenoside Rh2, which exerts the potent anticancer action both in vitro and in vivo, is one of the most well characterized ginsenosides extracted from ginseng. Although its effects on cancer are significant, the underlying mechanisms remain unknown. In this study, we sought to elucidate possible links between ginsenoside Rh2 and phosphoglucose isomerase/autocrine motility factor (PGI/AMF). Methods: $KG1{\alpha}$, a leukemia cell line highly expressing PGI/AMF was assessed by western blot analysis and reverse transcription- PCR (RT-PCR) assay after transfection of a small interfering (si)-RNA to silence PGI/AMF. The effect of PGI/AMF on proliferation was measured by typan blue assay and antibody array. A cell counting kit (CCK)-8 and flow cytometry (FCM) were adopted to investigate the effects of Rh2 on PGI/AMF. The relationships between PGI/AMF and Rh2 associated with Akt, mTOR, Raptor, Rag were detected by western blot analysis. Results: KG1${\alpha}$ cells expressed PGI/AMF and its down-regulation significantly inhibited proliferation. The antibody array indicated that the probable mechanism was reduced expression of PARP, State1, SAPK/JNK and Erk1/2, while those of PRAS40 and p38 were up-regulated. Silencing of PGI/AMF enhanced the sensibility of $KG1{\alpha}$ to Rh2 by suppressing the expression of mTOR, Raptor and Akt. Conclusion: These results suggested that ginsenoside Rh2 suppressed the proliferation of $KG1{\alpha}$, the same as down-regulation of PGI/AMF. Down-regulation of PGI/AMF enhanced the pharmacological effects of ginsenoside Rh2 on KG1${\alpha}$ by reducing Akt/mTOR signaling.

Reconstructed Adeno-Associated Virus with the Extracellular Domain of Murine PD-1 Induces Antitumor Immunity

  • Elhag, Osama A.O.;Hu, Xiao-Jing;Wen-Ying, Zhang;Li, Xiong;Yuan, Yong-Ze;Deng, Ling-Feng;Liu, De-Li;Liu, Ying-Le;Hui, Geng
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4031-4036
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    • 2012
  • Background: The negative signaling provided by interactions of the co-inhibitory molecule, programmed death-1 (PD-1), and its ligands, B7-H1 (PD-L1) and B7-DC (PD-L2), is a critical mechanism contributing to tumor evasion; blockade of this pathway has been proven to enhance cytotoxic activity and mediate antitumor therapy. Here we evaluated the anti-tumor efficacy of AAV-mediated delivery of the extracellular domain of murine PD-1 (sPD-1) to a tumor site. Material and Methods: An rAAV vector was constructed in which the expression of sPD-1, a known negative regulator of TCR signals, is driven by human cytomegalovirus immediate early promoter (CMV-P), using a triple plasmid transfection system. Tumor-bearing mice were then treated with the AAV/sPD1 construct and expression of sPD-1 in tumor tissues was determined by semi quantitative RT-PCR, and tumor weights and cytotoxic activity of splenocytes were measured. Results: Analysis of tumor homogenates revealed sPD-1 mRNA to be significantly overexpressed in rAAV/sPD-1 treated mice as compared with control levels. Its use for local gene therapy at the inoculation site of H22 hepatoma cells could inhibit tumor growth, also enhancing lysis of tumor cells by lymphocytes stimulated specifically with an antigen. In addition, PD-1 was also found expressed on the surfaces of activated CD8+ T cells. Conclusion: This study confirmed that expression of the soluble extracellular domain of PD-1 molecule could reduce tumor microenvironment inhibitory effects on T cells and enhance cytotoxicity. This suggests that it might be a potential target for development of therapies to augment T-cell responses in patients with malignancies.

A Performance Enhancement Scheme of Hierarchical Mobility Management in IPv6 Networks (IPv6 네트워크에서 계층적 이동성 관리의 성능향상 방안)

  • Seo, Jae-Kwon;Lee, Kyug-Geun
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.44 no.10
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    • pp.119-126
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    • 2007
  • Recently, the mobility of users and mobile communication technologies have developed rapidly. The users in this state also want to connect their devices and to receive services anywhere, anytime. Hierarchical Mobile IPv6 (HMIPv6) has been proposed by the Internet Engineering Task Force (IETF) to compensate for such problems as handover latency and signaling overhead when employing Mobile IPv6 (MIPv6). HMIPv6 supports micro-mobility within a domain and introduces a new entity, namely Mobility Anchor Point (MAP) as a local home agent. However, HMIPv6 has been found to cause longer handover latency when the inter-domain handover occurs. This is because a Mobile Node (MN) has to generate two addresses and register them to Home Agent (HA) a MAP, respectively. In order to solve such problems, we propose a scheme that an MN generates one address and registers it to HA for supporting fast handover during the inter-domain handover process. In the proposed scheme, the load of MAP and MAP domain is reduced because the number of MNs which are managed by MAP is decreased and the MAP does not perform proxy Neighbor Discovery Protocol (NDP) to intercept packets destined to MNs. We evaluate the performance of proposed scheme in comparison to HMIPv6 through the simulation and numerical analysis.

Analysis of germinating seed stage expressed sequence tags in Oryza sativa L. (벼 발아종자 발현유전자의 발현특성분석)

  • Yoon, Ung-Han;Lee, Gang-Seob;Kim, Chang-Kug;Lee, Jung-Sook;Hahn, Jang-Ho;Yun, Doh-Won;Ji, Hyeon-So;Lee, Tae-Ho;Lee, Jeong-Hwa;Park, Sung-Han;Kim, Gun-Wook;Seo, Mi-Suk;Kim, Yong-Hwan
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.281-288
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    • 2009
  • Seed germination is the important stage to express many genes for regulation of energy metabolism, starch degradation and cell division from seed dormancy state. For the functional analysis of seed germination mechanisms, we were analyzed the rice cDNA clones (Oryzasativa cultivar Ilpum) obtained from seed imbibition during 48 hours. Total number of 18,101 Expressed Sequence Tags (ESTs) were clustered using SeqMan program. Among them, 8,836 clones were identified as unique clones. We identified the chitinase gene specifically expressed in seed germination and amylase gene involved to starch degradation from the full length cDNA analysis, and several genes were registered to NCBI GeneBank. To analyzed the commonly expressed genes between inmature seed and germinated seed, 25,66 inmature ESTs and 18,101 germinated ESTs were clustered using SeqMan program and identified 2,514 clones as commonly expressed unigene. Among them, alpha-glubulin and alcohol dehydrogenase I were supposed to LEA genes only expressed in the immature and germinated seed stages. For the clustering of orthologous group genes, we further analyzed the 8,836 EST clones from germinating seeds using NCBI clusters of orthologous groups database. Among the clones, 5,076 clones were categorized into information storage and processing, cellular processes and signaling, metabolism and poorly characterized genes, proportioning 783 (14.29%), 1,484 (27%), 1,363 (24.8%) and 1,869 (34%) clones to the previous four categories, respectively.

An Authentication and Handoff Mechanism using AAA and HMIPv6 on NEMO Environment (이동 네트워크(NEMO)에서 HMIPv6를 적용한 AAA 인증 방안 연구)

  • Choi, Kyung;Kim, Mi-Hui;Chae, Ki-Joon
    • The KIPS Transactions:PartC
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    • v.16C no.2
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    • pp.165-182
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    • 2009
  • Mobile IPv6 spends considerable bandwidth considering that its signal volume is proportional to the mobile and also it should be strengthened to support the binding signal volume, the traffic, and effective mobility. So, the study in NEMO(Network Mobility), an extended version of Mobile IPv6, has been conducted. NEMO provides its mobility by putting several mobiles and more than one portable router into one unit called as mobile network. Because nodes access Internet via the portable router at this time, it receives transparency without any additional work and that much reduces binding signal while solving binding storm. By supporting mobility, NEMO is able to have various mobile structures which realize several networks hierarchically and it is necessary to improve its safety and security by authenticating among the upper networks or the lower ones while moving. Also, it is extremely required to begin a study in the device to improve efficiency accompanied with mobility, which is executed by the fast hand-off as well as the safe authentication. For those reasons, this paper not only classifies various NEMO mobile scenarios into 7 ways, but also provides AAA authentication of each scenario, the authentication through the safety authentication and fast handoff authentication using F+HMIPv6 and the way to reduce both signaling volume and packet delays efficiently during the handoff.