• Title/Summary/Keyword: multiple-RAT

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Cloning of Xenopus laevis TRPV2 by Gene Prediction

  • Lee, Jung Youn;Shim, Won Sik;Oh, Uhtaek
    • Genomics & Informatics
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    • v.3 no.1
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    • pp.24-29
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    • 2005
  • TRPV2 is a non-specific cation channel expressed in sensory neurons, and activated by noxious heat. Particularly, TRPV2 has six transmembrane domains and three ankyrin repeats. TRPV2 has been cloned from various species such as human, rat, and mouse. Oocytes of Xenopus laevis - an African clawed frog ­have been widely used for decades in characterization of various receptors and ion channels. The functional property of rat TRPV2 was also identified by this oocyte expression system. However, no TRPV2 orthologue of Xenopus laevis has been reported so far. Hence, we have focused to clone a TRPV2 orthologue of Xenopus laevis with the aid of bioinformatic tools. Because the genome sequence of Xenopus laevis is not available until now, a genome sequence of Xenopus tropicalis - a close relative species of Xenopus laevis - was used. After a number of bioinformatic searches in silico, a predicted full-length sequence of TRPV2 orthologue of Xenopus tropicalis was found. Based on this predicted sequence, various approaches such as RT-PCR and 5' -RACE technique were applied to clone a full length of Xenopus laevis TRV2. Consequently, a full-length Xenopus laevis TRPV2 was cloned from heart cDNA.

Calumenin Interacts with SERCA2 in Rat Cardiac Sarcoplasmic Reticulum

  • Sahoo, Sanjaya Kumar;Kim, Do Han
    • Molecules and Cells
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    • v.26 no.3
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    • pp.265-269
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    • 2008
  • Calumenin, a multiple EF-hand $Ca^{2+}$ binding protein is located in the SR of mammalian heart, but the functional role of the protein in the heart is unknown. In the present study, an adenovirus gene transfer system was employed for neonatal rat heart to examine the effects of calumenin over-expression (Calu-OE) on $Ca^{2+}$ transients. Calu-OE (8 folds) did not alter the expression levels of DHPR, RyR2, NCX, SERCA2, CSQ and PLN. However, Calu-OE affected several parameters of $Ca^{2+}$ transients. Among them, prolongation of time to 50% baseline ($T_{50}$) was the most outstanding change in electrically-evoked $Ca^{2+}$ transients. The higher $T_{50}$ was due to an inhibition of SERCA2-mediated $Ca^{2+}$ uptake into SR, as tested by oxalate-supported $Ca^{2+}$ uptake. Furthermore, co-IP study showed a direct interaction between calumenin and SERCA2. Taken together, calumenin in the cardiac SR may play an important role in the regulation of $Ca^{2+}$ uptake during the EC coupling process.

Rapid Quantification of Topotecan in Biological Samples by Liquid Chromatography/Tandem Mass Spectrometry

  • Shin, Beom-Soo;Lee, Mann-Hyung;Yoo, Sun-Dong
    • Journal of Pharmaceutical Investigation
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    • v.39 no.5
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    • pp.367-372
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    • 2009
  • A rapid liquid chromatography/tandem mass spectrometry (LC/MS/MS) assay method was developed for the determination of topotecan levels in rat serum. The assay utilized a single liquid-liquid extraction with a mixture of ethy l acetate and acetonitrile (6:1 v/v) and isocratic elution. The multiple reaction monitoring was based on the transition of m/z 422.0$\rightarrow$376.5 for topotecan and 315.1$\rightarrow$226.6 for clomipramine (internal standard). The developed assay was validated to demonstrate the specificity, recovery, lower limit of quantification (LLOQ), accuracy and precision. The assay was linear over a concentration range from 0.5-100 ng/mL, with LLOQ being 0.5 ng/mL using a small volume of rat serum (0.1 mL). The mean intra- and inter-day assay accuracy was 87.7-111.0% and 97.8-108.3, respectively, and the mean intra- and interday precision was between 1.6-4.3% and 3.8-10.3, respectively. The developed assay was applied to a pharmacokinetic study after a bolus i.v. injection of topotecan in rats.

Age-dependent expression of ion channel genes in rat

  • Sung-Cherl Jung;Tong Zhou;Eun-A Ko
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.1
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    • pp.85-94
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    • 2023
  • Ion channels regulate a large number of cellular functions and their functional role in many diseases makes them potential therapeutic targets. Given their diverse distribution across multiple organs, the roles of ion channels, particularly in age-associated transcriptomic changes in specific organs, are yet to be fully revealed. Using RNA-seq data, we investigated the rat transcriptomic profiles of ion channel genes across 11 organs/tissues and 4 developmental stages in both sexes of Fischer 344 rats and identify tissue-specific and age-dependent changes in ion channel gene expression. Organ-enriched ion channel genes were identified. In particular, the brain showed higher tissue-specificity of ion channel genes, including Gabrd, Gabra6, Gabrg2, Grin2a, and Grin2b. Notably, age-dependent changes in ion channel gene expression were prominently observed in the thymus, including in Aqp1, Clcn4, Hvcn1, Itpr1, Kcng2, Kcnj11, Kcnn3, and Trpm2. Our comprehensive study of ion channel gene expression will serve as a primary resource for biological studies of aging-related diseases caused by abnormal ion channel functions.

Gene Expression Analysis of Rat Liver Epithelial Cells in Response to Thioacetamide

  • Park, Joon-Suk;Yeom, Hye-Jung;Jung, Jin-Wook;Hwang, Seung-Yong;Lee, Yong-Soon;Kang, Kyung-Sun
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.203-208
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    • 2005
  • Thioacetamide (TA) is potent haptotoxincant that requires metabolic activation by mixed-function oxidases. Micrcarray technology, which is massive parallel gene expression profiling in a single hybridization experiment, has provided as a powerful molecular genetic tool for biological system related toxicant. In this study we focus on the use of toxicogenomics for the determination of gene expression analysis associated with hepatotoxicity in rat liver epithelial cell line WB-F344 (WB). The WB cells was used to assess the toxic effects of TA. WB cells were exposed to two concentrations of TA-doses which caused 20% and 50% cell death were chosen and the cells exposed for periods of 2 and 24 h. Our data revealed that following the 2-h exposure at the both of doses and 24-h exposure at the low doses, few changes in gene expression were detected. However, after 24-h exposure of the cells to the high concentration, multiple changes in gene expression were observed. TA treatment gave rise predominantly to up-regulation of genes involved in cell cycle and cell death, but down-regulation of genes involves in cell adhesion and calcium ion binding. Exposure of WB cells to higher doses of the TA gave rise to more changes in gene expression at lower exposure times. These results show that TA regulates expression of numerous genes via direct molecular signaling mechanisms in liver cells.

Expression of Ski in the Corpus Luteum in the Rat Ovary

  • Kim, Hyun;Matsuwaki, Takashi;Yamanouchi, Keitaro;Nishihara, Masugi;Yang, Boh-Suk;Ko, Yeoung-Gyu;Kim, Sung-Woo
    • Journal of Embryo Transfer
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    • v.26 no.4
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    • pp.229-235
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    • 2011
  • Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. Ski protein is implicated in proliferation/differentiation in a variety of cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of this study was, by means of immunohistochemical methods, to locate Ski protein in the rat ovaries during ovulation and corpora lutea (CL) formation to predict the possible involvement of Ski in luteinization. In addition, we performed to examine whether the initiation of luteinization with luteinizing hormone (LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vivo models. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rat, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone (LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of corpus luteum (CL). These results indicate that Ski is profoundly expressed in the luteinized granulosa cells and luteal cells of CL during luteinization, and suggest that Ski may play a role in luteinization of granulosa cells.

Molecular Identification and Distribution of Aquaporins in Human and Rat Testes (사람과 흰쥐의 고환에서 Aquaporin 유전자의 발현)

  • Park, Nam-Cheol;Park, Young-Soo;Oh, Gom-Su;Jung, Jin-Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.2
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    • pp.133-144
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    • 2000
  • Objective: Several water channels (aquaporins; AQP) that belong to the MIP (major intrinsic protein) family have identified. In the selected tissues including red blood cells or renal tubules, water movements are abundant and/or physiologically important. Unexpectedly, a high water permeability of human and ram sperm has been reported. Recent studies showed that AQP7 and AQP8 are present in testes, so that the high water permeability of human sperm suggested to be mediated by AQPs. Method: To identify the identity of aquaporins expressed in testes, RT-PCR was performed using degenerative primers, which were designed to correspond to highly conserved sequences surrounding the Asn-Pro-Ala (NPA) motifs in the aquaporins. New expressed AQP series were reconfirmed by immunohistochemical study using rabbit polyclonal antibodies. Results: DNA sequencing of PCR products revealed that AQP2 and AQP3 mRNA as well as AQP7 and AQP8 are expressed in human and rat testes. In human and rat testes, AQP2 are expressed in spermatozoa, interstitial cells and myofibroblasts and AQP3 are expressed in myofibroblasts of semineferous tubules on immunocytochemical stain. Conclusion: These results indicate that multiple aquaporins are expressed in testes, and that they may have important roles in the spermatogenesis and the germ cell function of testis.

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Development and Validation of an LC-MS/MS Method for Determination of Damaurone D in Rat Plasma and its Application to Pharmacokinetic Study in Rats

  • Lee, Wonpyo;Song, Im-Sook;Han, Young Taek;Choi, Min-Koo
    • Mass Spectrometry Letters
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    • v.10 no.1
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    • pp.38-42
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    • 2019
  • Damaurone D belongs to the genus Rosa and is a traditional medicinal product used for the treatment of depression, inflammation, and infectious diseases. The purpose of this study was to develop a simple liquid chromatography-tandem mass spectrometry method for the detection of damaurone D in rat plasma and to demonstrate its application in pharmacokinetic studies. Damaurone D and berberine (internal standard) were extracted with acetonitrile using a protein precipitation method. Mass transition was monitored in multiple reaction monitoring mode at m/z $323.2{\rightarrow}267.0$ for damaurone D and m/z $336.1{\rightarrow}320.0$ for berberine in positive ion mode. Analytical validation was conducted by evaluating the specificity, linearity, accuracy, precision, matrix effect, extraction recovery, and stability. The calibration curves were linear over 2-1000 ng/mL. The intra- and inter-day precision and accuracy of quality control samples were 4.79-13.33% and 86.23-102.75%, respectively. The matrix effect and extraction recovery were 96.11-98.47% and 96.11-102.25%, respectively. In the pharmacokinetic study after intravenous administration of damaurone D at a dose of 3 mg/kg in rats, the area under the curve and clearance of damaurone D in rat plasma were $16750.26{\pm}2676.10min{\cdot}ng/mL$ and $182.44{\pm}31.36mL/min/kg$, respectively.

A Mobile P2P Message Platform Enabling the Energy-Efficient Handover between Heterogeneous Networks (이종 네트워크 간 에너지 효율적인 핸드오버를 지원하는 모바일 P2P 메시지 플랫폼)

  • Kim, Tae-Yong;Kang, Kyung-Ran;Cho, Young-Jong
    • Journal of KIISE:Computing Practices and Letters
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    • v.15 no.10
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    • pp.724-739
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    • 2009
  • This paper suggests the energy-efficient message delivery scheme and the software platform which exploits the multiple network interfaces of the mobile terminals and GPS in the current mobile devices. The mobile terminals determine the delivery method among 'direct', 'indirect', and 'WAN' based on the position information of itself and other terminals. 'Direct' method sends a message directly to the target terminal using local RAT. 'Indirect' method extends the service area by exploiting intermediate terminals as relay node. If the target terminal is too far to reach through 'direct' or 'indirect' method, the message is sent using wireless WAN technology. Our proposed scheme exploits the position information and, thus, power consumption is drastically reduced in determining handover time and direction. Network simulation results show that our proposed delivery scheme improves the message transfer efficiency and the handover detection latency. We implemented a message platform in a smart phone realizing the proposed delivery scheme. We compared our platform with other typical message platforms from energy efficiency aspect by observing the real power consumption and applying the mathematical modeling. The comparison results show that our platform requires significantly less power.

A Study of Development for High-speed Cloud Video Service using SDN based Multi Radio Access Technology Control Methods (초고속 클라우드 비디오 서비스 실현을 위한 SDN 기반의 다중 무선접속 기술 제어에 관한 연구)

  • Kim, Dongha;Lee, Sungwon
    • Journal of Broadcast Engineering
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    • v.19 no.1
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    • pp.14-23
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    • 2014
  • This paper proposed controlling methods for SDN(Software Defined Network) based multiple radio access technology as the solutions of following two issues which were mainly occurred by explosive increasing of video traffic. The first one is a requirement for traffic off-loading caused by 3rd-party video service providers from the mobile network operator's viewpoint. The other one is a provision of high-speed video contents transmission services with low price. Furthermore, the performance evaluation was also conducted on the real test-bed which is composed of OpenStack cloud and SDN technology such as OpenFlow and Open vSwitch. A virtual machine running on the OpenStack provide a video service and the terminal which is able to use multiple radio access technology supports two 2.4GHz WLANs(Wireless Local Area Network) and three 5GHz WLANs, concurrently. Finally, we can get 820Mbps of the maximum transmission speed by using that five WLAN links for the single service at the same time.