• Title/Summary/Keyword: Reverse annealing

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Identification of Differentially Expressed Genes by Gabapentin in Cultured Dorsal Root Ganglion in a Rat Neuropathic Pain Model

  • Heo, Ji Hye;Lee, Seung Ha;Chang, Kyung Ha;Han, Eun Hye;Lee, Seung Gwan;Choi, Dal Woong;Kim, Suhng Wook
    • Biomolecules & Therapeutics
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    • v.21 no.2
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    • pp.126-131
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    • 2013
  • Neuropathic pain is a chronic pain disorder caused by nervous system lesions as a direct consequence of a lesion or by disease of the portions of the nervous system that normally signal pain. The spinal nerve ligation (SNL) model in rats that reflect some components of clinical pain have played a crucial role in the understanding of neuropathic pain. To investigate the direct effects of gabapentin on differential gene expression in cultured dorsal root ganglion (DRG) cells of SNL model rats, we performed a differential display reverse transcription-polymerase chain reaction analysis with random priming approach using annealing control primer. Genes encoding metallothionein 1a, transforming growth factor-${\beta}1$ and palmitoyl-protein thioesterase-2 were up-regulated in gabapentin-treated DRG cells of SNL model rats. The functional roles of these differentially expressed genes were previously suggested as neuroprotective genes. Further study of these genes is expected to reveal potential targets of gabapentin.

Identification of Differentially Expressed Genes between Neonatal and Peripubertal Rat Thymi Using $GeneFishing^{TM}$ Polymerase Chain Reaction

  • Kang, Da-Won;Kim, Gyu-Tae;Han, Jae-Hee
    • Reproductive and Developmental Biology
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    • v.31 no.1
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    • pp.55-60
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    • 2007
  • Aging causes thymus involution, and genes in thymus play an important role in the development of the immune system. In this study, we compared genes expressed in thymus of neonatal and peripubertal rats using annealing control primers (ACPs)-based GeneFishing polymerase chain reaction (PCR) and semiquantitative reverse transcription (RT)-PCR. We identified 10 differentially expressed genes (DEGs) with 20 ACPs. Of 10 DEGs, bystin-like, collagen type V alpha 1 (COL5A1), and T-cell receptor beta-chain segment 2 (TCRB2) that are related to immune-function were detected in rat thymus. Bystin-like and TCRB2 were up-regulated, while COL5A1 was down-regulated in peripubertal thymus. Semiquantitative RT-PCR confirmed postnatal changes in expression of bystin-like, COL5A1, and TCRB2. These results suggest that bystin-like, COL5A1, and TCRB2 could regulate immune function controlled in thymus as age increases.

Differentially Expressed Genes in Porcine Parthenogenetic 2-cell and Blastocyst Embryos

  • Lee, Hwa-young;Kim, Bong-Ki;Cui, Xiang-Chun;Hwang, Kyu-Chan;Chung, Hak-Jae;Ko, Yeoung-Gyu;Jung, Jin-Kwan;Chang, Won-Kyong;Kim, N. H.
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.198-198
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    • 2004
  • The identification of embryo-specific genes would provide insights into early embryonic development. However, the current methods employed to identify the genes that are expressed at a specific developmental stage are labor intensive and suffer from high rates of false positives. Here we employed a new and accurate reverse transcription-polymerase chain reaction (RT-PCR) technology that involves annealing control primers(ACPs) to identify the genes that are specifically expressed in porcine blastocysts compared to 2-cell stage embryos. (omitted)

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Application of Bovine Viral Diarrhoea Virus as an Internal Control in Nucleic Acid Amplification Tests for Hepatitis C Virus RNA in Plasma-Derived Products

  • Yoo Si Hyung;Hong Seung Hee;Jung Sa Rah;Park Su Jin;Lee Nam Kyung;Kim Soon Nam;Kang Sang Mo;Min Hong Ki;Park Sue Nie;Hong Seung Hwa
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.72-76
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    • 2006
  • Plasma-derived products are produced from plasma via fractionation and chromatography techniques, but can also be produced by other methods. In the performance of nucleic acid amplification tests (NAT) with plasma-derived products, it is necessary to include an internal control for the monitoring of all procedures. In order to avoid false negative results, we confirmed the usefulness of the bovine viral diarrhoea virus (BVDV) for use as an internal control in the detection of hepatitis C virus (HCV) RNA in plasma-derived products. These products, which were spiked with BVDV, were extracted and then NAT was performed. Specificity and sensitivity were determined via the adjustment of primer concentrations and annealing temperatures. BVDV detection allows for validation in the extraction, reverse transcription, and amplification techniques used for HCV detection in plasma-derived products.

Arsenic-Induced Differentially Expressed Genes Identified in Medicago sativa L. roots

  • Rahman, Md. Atikur;Lee, Sang-Hoon;Kim, Ki-Yong;Park, Hyung Soo;Hwang, Tae Young;Choi, Gi Jun;Lee, Ki-Won
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.3
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    • pp.243-247
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    • 2016
  • Arsenic (As) is a toxic element that easily taken up by plants root. Several toxic forms of As disrupt plant metabolism by a series of cellular alterations. In this study, we applied annealing control primer (ACP)-based reverse transcriptase PCR (polymerase chain reaction) technique to identify differentially expressed genes (DEGs) in alfalfa roots in response to As stress. Two-week-old alfalfa seedlings were exposed to As treatment for 6 hours. DEGs were screened from As treated samples using the ACP-based technique. A total of six DEGs including heat shock protein, HSP 23, plastocyanin-like domain protein162, thioredoxin H-type 1 protein, protein MKS1, and NAD(P)H dehydrogenase B2 were identified in alfalfa roots under As stress. These genes have putative functions in abiotic stress homeostasis, antioxidant activity, and plant defense. These identified genes would be useful to increase As tolerance in alfalfa plants.

Ultra shallow $p^{+}$n junction formation using the boron diffusin form epi-co silicide (에피 코발트 실리사이드막으로 부터의 붕소 확산을 이용한 극저층 $p^{+}$n 접합 형성)

  • 변성자;권상직;김기범;백홍구
    • Journal of the Korean Institute of Telematics and Electronics A
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    • v.33A no.7
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    • pp.134-142
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    • 1996
  • The epi-CoSi$_{2}$ layer was formed by alloying a Co(120$\AA$)/Ti(50$\AA$) bilayer. In addition, the ultra shallow p$^{+}$n junction of which depth is about not more than 40nm at the background concentration, 10$^{18}$atoms/cm$^{3}$ could be formed by annealing (RTA-II) the ion implanted epi-silicide. When the temperature of RTA-I is as low as possible and that of RTA-II is moderate, the p$^{+}$n junction that has low leakage current and stable epi-silicide layer could be obtained. That is, when th econdition of TRA-I was 900$^{\circ}C$/20sec and that of RTA-II was 900$^{\circ}C$/10sec, the reverse leakage current was as high as 11.3$\mu$A/cm$^{2}$ at -5V. The surface of CoSi$_{2}$ appeared considerably rough. However, when the conditon of RTA-I was 800$^{\circ}C$/20sec or 700$^{\circ}C$/20sec, the leakage currents were as low as 8.3nA/cm$^{2}$ and 9.3nA/cm$^{2}$, respectively and also the surfaces appeared very uniform.

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A Study on the Crystallization of Grain-Boundary Phases in Si3N4-Y2O3-Al2O3 System (Si3N4-Y2O3-Al2O3계의 입계상 결정화에 관한 연구)

  • 박정현;황종희
    • Journal of the Korean Ceramic Society
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    • v.26 no.1
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    • pp.13-20
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    • 1989
  • After sintering Si3N4 containing 20wt% of variable composition ratio of Y2O3 and Al2O3 at 1$600^{\circ}C$, the specimens were annealed at 125$0^{\circ}C$ and 135$0^{\circ}C$ for 5, 10, 15 hours in order to crystallize the remanining oxynitride glass phases. The main grain-boundary crystalline phases in the Si3N4-Y2O3-Al2O3 system were melilite and YAG. By annealing 15hrs. at 125$0^{\circ}C$, almost all of the glasses were crystallized. During the growth of melilite, lattice volyume of $\beta$-Si3N4 was increased as Al3+ and O2- ions in the oxynitride glass diffuse into $\beta$-Si3N4 lattice, but during the growth of YAG, lattice volume of $\beta$-Si3N4 was decreased by reverse diffusion of Al3+ and O2- ions. In case of crystallization of glass phase to melilite, thermal expansion of sample was decreased, but in case of crystallization to YAG, inverse phenomen on was observed.

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Identification of Differentially Expressed Genes in Human Mesenchymal Stem Cell-Derived Neurons

  • Heo, Ji-Hye;Cho, Kyung-Jin;Choi, Dal-Woong;Kim, Suhng-Wook
    • Toxicological Research
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    • v.26 no.1
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    • pp.15-19
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    • 2010
  • Mesenchymal stem cells (MSCs) have greater potential for immediate clinical and toxicological applications, due to their ability to self-renew, proliferate, and differentiate into a variety of cell types. To identify novel candidate genes that were specifically expressed during transdifferentiation of human MSCs to neuronal cells, we performed a differential expression analysis with random priming approach using annealing control primer-based differential display reverse transcription-polymerase chain reaction approach. We identified genes for acyl-CoA thioesterase, tissue inhibitor of metalloproteinases-1, brain glycogen phosphorylase, ubiquitin C-terminal hydrolase and aldehyde reductase were up-regualted, whereas genes for transgelin and heparan sulfate proteoglycan were down-regulated in MSC-derived neurons. These differentially expressed genes may have potential role in regulation of neurogenesis. This study could be applied to environmental toxicology in the field of testing the toxicity of a chemical or a physical agent.

Gene Expression Analysis of Pregnant Specific Stage in the Miniature Pig Ovary

  • Yun, Seong-Jo;Noh, Won-Gun;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.249-255
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    • 2009
  • The miniature pig is considered to be a better organ donor breed for xenotransplantation than other pig breeds because the size of the organs of the miniature pig is similar to that of humans. In this study, we aimed at identifying differentially expressed genes in the miniature pig ovary during pregnancy. For this, we used the miniature pig ovary model, annealing control primer-based reverse transcription polymerase chain reaction (PCR), quantitative real-time PCR (qRT-PCR), and northern blotting analysis. We identified 13 genes showing differential expression on the based of pregnancy status and validated 8 genes using qRT-PCR. We also sequenced the full-length cDNA of ephrin receptor A4 (EphA4), which had a significant difference in expression level, and validated it by northern blotting. These genes may provide a better understanding of the cellular and molecular mechanisms during pregnancy in miniature pig ovary.

Self-Aligned $n^+$ -pPolysilicon-Silicon Junction Structure Using the Recess Oxidation (Recess 산화를 이용한 자기정렬 $n^+$ -p 폴리실리콘-실리콘 접합구조)

  • 이종호;박영준;이종덕;허창수
    • Journal of the Korean Institute of Telematics and Electronics A
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    • v.30A no.6
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    • pp.38-48
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    • 1993
  • A recessed n-p Juction diode with the self-aligned sturcture is proposed and fabricated by using the polysilicon as an n$^{+}$ diffusion source. The diode structure can be applicable to the emitter-base formation of high performance bipolar divice and the n$^{+}$ polysilicone mitter has an important effect on the device characteristics. The considered parameters for the polysilicon formation are the deposition condition. As$^{+}$ dose for the doping of the polysilicon and the annealing condition using RTP system. The vertical depth profiles of the fabricated diode are obtained by SIMS and the electrical characteristics are analyzed in terms of the ideality factor of diode (n), contact resistance and reverse leakage current. In addition, n$^{+}$-p junction diodes are formed by using the amorphous silicon (of combination of amorphous and polysiliocn) instead of polysilicon and their characteristics are compared with those of the standard sample. The As$^{+}$ dose for the formation of good junction is about 1~2${\times}10^{16}cm^{2}$ at given RTA conditions (1100.deg. C, 10sec).

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