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Endothelin-l as a Regulator of Vascular Smooth Muscle Contraction-related Signal Transduction and Endothelin-l-induced Pain-related Nociception -The Approach of Basic Physical Therapy for the Study of Pain Specialized Physical Therapy- (혈관평활근 수축-연관 신호전달 체계에 대한 Endothelin-1의 역할과 Endothelin-1-유도통증-연관 유해감각 -통증전문물리치료 연구를 위한 기초물리치료학적 접근을 중심으로-)

  • Kim, Jung-Hwan;Lee, Sook-Hee;Lee, Sang-Bin;Choi, Yoo-Rim;Kim, Bo-Kyung;Park, Ju-Hyun;Koo, Ja-Pung;Choi, Wan-Suk;An, Ho-Jung;Choi, Jeong-Hyun;Kim, Moo-Gi;Kim, Soon-Hee
    • Journal of Korean Physical Therapy Science
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    • v.13 no.2
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    • pp.99-119
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    • 2006
  • Endothelin (ET) is a 21 amino acid peptide with multifunctional effects on the vasculature as well as a variety of other cell types such as respiratory, gastrointestinal, urogenital, endocrine, central nervous systems, and others. Endothelin has emerged as a modulator by autocrine and paracrine actions for many cellular activities, including vasoconstriction, cell proliferation, hormone production, neurotransmitter and/or neuromodulator. The endothelin family consists of three closely related peptides, ET-1, ET-2, and ET-3 derived from separate genes, such as chromosome 6, 1, and 20, respectively. ET-1 is the predominant isoform produced in the cardiovascular system and about which most is known. Endothelin receptors are seven-transmembrane GTP-binding protein-coupled receptors, which are classified into endothelin-A (ETA) and endothelin-B (ETB) receptors. Interestingly, recent evidence is accumulating to suggest that ET -1 may contribute to a variety of pain states such as allodynia and hyperalgesia in animals and humans. Therefore, in this review the biological characteristics and contraction-related mechanism of endothelin-1 in mammalian cells will be summarized. Especially, we focus on multifunctional roles for ET-1 in noxious stimulation-induced pain for the study of pain specialized physical therapy.

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c-Jun N-terminal Kinase Contributes to Norepinephrine-Induced Contraction Through Phosphorylation of Caldesmon in Rat Aortic Smooth Muscle

  • Lee, Youn-Ri;Lee, Chang-Kwon;Park, Hyo-Jun;Kim, Hyo-Jin;Kim, Jung-Hwan;Kim, Jae-Heung;Lee, Keun-Sang;Lee, Yun-Lyul;Min, Kyung-Ok;Kim, Bo-Kyung
    • Journal of Korean Physical Therapy Science
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    • v.13 no.2
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    • pp.129-135
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    • 2006
  • Vascular smooth muscle contraction is mediated by activation of extracellular signal-regulated kinase (ERK) 1/2, an isoform of mitogen-activated protein kinase (MAPK). However, the role of stress-activated protein kinase/c-Jun N-terminal kinase (JNK) in vascular smooth muscle contraction has not been defined. We investigated the role of JNK in the contractile response to norepinephrine (NE) in rat aortic smooth muscle. NE evoked contraction in a dose-dependent manner, and this effect was inhibited by the JNK inhibitor SP600125. NE increased the phosphorylation of JNK, which was greater in aortic smooth muscle from hypertensive rats than from normotensive rats. NE-induced JNK phosphorylation was significantly inhibited by SP600125 and the conventional-type PKC (cPKC) inhibitor Go6976, but not by the Rho kinase inhibitor Y27632 or the phosphatidylinositol 3-kinase inhibitor LY294002. Thymeleatoxin, a selective activator of cPKC, increased JNK phosphorylation, which was inhibited by $G{\ddot{o}}6976$. SP600125 attenuated the phosphorylation of caldesmon, an actin-binding protein whose phosphorylation is increased by NE. These results show that JNK contributes to NE-mediated contraction through phosphorylation of caldesmon in rat aortic smooth muscle, and that this effect is regulated by the PKC pathway, especially cPKC.

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Analysis of Protein Function and Comparison on Expression of Protein in Taekwang During Maturation using Proteomic Techniques (Proteomics를 이용한 등숙기 차이에 따른 콩 종실 저장단백질 발현양상 비교 분석)

  • Cho, Seong-Woo;Kim, Tae-Sun;Kwon, Soo-Jeong;Roy, Swapan Kumar;Lee, Chul-Won;Kim, Hong-Sig;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.41-46
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    • 2015
  • In the present study, different expression of protein from Taekwang was revealed by 2-DE, and expressions of protein on each week after flowering was investigated. After analysis of expression of protein, MALDI-TOF was executed to identify expected protein function. Results revealed that there were three patterns of expression of protein during the maturing. The first pattern was that proteins were gradually expressed as up-regulation from 1 week to 6 week. The second pattern was that proteins were expressed gradually from 1 week to 5 week and then it started down-regulation in 6 week. The last pattern was that proteins were gradually as up-regulation from 1 week to 3 week and then down-regulation until 6 week. This phenomenon suggests that young stage has more protein related to correspondence mechanism against disease and growth and then maturing stage has more expression of protein related to storage protein. In MALDI-TOF analysis, p24 oleosin isoform A protein was identified that relates oleosin which is synthetic product in oil body. This protein spot increased gradually until 5 week and then decreased after 5 week. It explained that the protein is active until maturing stage to protect oil in seed and then its activity has gradually degraded. This result may be expected that a protein, related to growth of a seed has increased until maturing and then a seed fills up with a storage protein.

Characterization of Protein Function and Differential Protein Expression in Soybean under Soaking Condition (Proteomics를 이용한 콩의 발아 전 침종처리에 따른 단백질 발현 양상 비교 분석)

  • Cho, Seong-Woo;Kim, Tae-Sun;Kwon, Soo-Jeong;Roy, Swapan Kumar;Lee, Chul-Won;Kim, Hong-Sig;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.114-122
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    • 2015
  • Soybean is very useful crop to supply vegetable protein for human. However, cultivation arear of this economically important crop is gradually diminished in upland field. Hence, cultivation area of soybean is increased in paddy field. During the growth duration of soybean, excessive moisture injury is serious problem for sustainable production and supply. We investigated protein expression according to different period of seed soaking and germination after seed soaking. For comparison on expression of protein according to different condition, we performed two-dimensional electrophoresis. After electrophoresis analysis, we selected differentially expressed protein spots according to different condition such as soaking period and germination after soaking to identify protein function by using MALDI-TOF. Results revealed that pattern of expression of protein according to soaking period and germination after soaking were generally not different in major spots. However, degree of expression of protein in some protein spots was increased in accordance with decrease of soaking period. Especially, in Hwangkeum-Kong, Danyeop-Kon, and Pecking, the degree of expression of protein was remarkably increased for 4 days after soaking. But, according to germination after soaking, degree of expression of protein in germinated seeds of all cultivars was higher than un-germinated seeds. In results of MALDI-TOF analysis, specific proteins were identified by different soaking period such as Allergen Gly m Bd 28K, P24 oleosin isoform B. Also, in accordance with germination, degree of protein expression of the related protein, Gibberellin was increased in un-germinated seeds of Iksan-Kong. In ungerminated seeds of Sinpaldal-kong, proteins were identified as down-regulated by soaking such as ATP binding and Inhibitor II', proteinase.

The Association of Interleukin-1 Receptor Antagonist and Peroxisome Proliferator-Activated $Receptor-{\gamma}2$ Pro12Ala Polymorphism with Ischemic Stroke in Type 2 Diabetes Mellitus Patients. (제2형 당뇨병 환자에서 허혈성 뇌졸중 발생과 Interleukin-1 Receptor Antagonist 및 Peroxisome Proliferator-Activated $Receptor-{\gamma}2$ 유전자 다형성과의 관계)

  • Jang, Won-Man;Lee, Byung-Cheol;Ahn, Se-Young;Doo, Ho-Kyung;Ahn, Young-Min
    • The Journal of Internal Korean Medicine
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    • v.28 no.2
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    • pp.262-274
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    • 2007
  • Objective : It has been reported that two-repeats ($IL1RN^{\ast}2$) of interleukin-1 receptor antagonist (IL-1Ra) gene is associated with ischemic stroke, and that Ala allele of the common Pro12Ala polymorphism in $PPAR-{\gamma}2$ isoform is associated with reduced risk for type 2 DM and its complications. The aim of the present study is to assess the association of IL-1Ra and $PPAR-{\gamma}2$ Pro12Ala polymorphism with the presence of ischemic stroke in the case of diabetic and non-diabetic patients. Methods : Genomic DNA was obtained from 373 healthy subjects, 157 DM subjects without ischemic stroke (known DM duration ${\ge}10$ years) and 302 ischemic stroke patients (including with DM). IL-1Ra polymorphism was analysed by polymerase chain reaction (PCR), and $PPAR-{\gamma}2$ polymorphism by restriction fragment length polymorphism after PCR. Results : $IL1RN^{\ast}1/IL1RN^{\ast}2$ genotype was associated with significantly increased risk for DM (OR=2.86, P = 0.0008) and ischemic stroke (OR=2.74, P = 0.0016). Pro/Ala genotype was associated with the reduced risk for DM (OR=0.53, P = 0.0491) and ischemic stroke (OR=0.38, P = 0.0039). They were also associated with the reduced risk for ischemic stroke in the DM patients compared with DM without ischemic stroke (OR=0.25, P = 0.0321). Conclusions : $IL1RN^{\ast}2$ allele could be an accelerating factor, not a predictive marker for ischemic stroke in type 2 DM. The Pro/Ala genotype of $PPAR-{\gamma}2$ Pro12Ala polymorphism may be associated with reduced risk for ischemic stroke with type 2 DM. Therefore it could be a useful predictive marker for ischemic stroke in Korean type 2 DM.

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Analysis of the Correlation between Expressions of HSP90α, HSP90β, and GRP94, and the Clinicopathologic Characteristics in Tissues of Non-Small Cell Lung Cancer Patients (비소세포 폐암 환자 조직에서 Hsp90α, Hsp90β, GRP94의 발현과 임상병리학적 특성과의 상관관계 분석)

  • Kim, Mi Kyeong
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.460-469
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    • 2017
  • Heat shock proteins (HSPs) are induced as a self-defense mechanism of cells when exposed to various external stresses, such as high fever, infection, free radicals, and heavy metals. They affect the prognosis in the process of tumor formation. HSP is classified into four families: HSP27, HSP60, HSP90, and HSP100, depending on molecular weight. Heat shock protein 90 (HSP90), a molecular chaperone, plays an important role in the cellular protection against various stressful stimuli and in the regulation of cell cycle progression and apoptosis. In the present study, we assessed the differential expression of HSP90 family proteins in non-small cell lung cancer (NSCLC), and the correlation of their expression levels with clinicopathologic factors and patient survival rates. The result of this study can be summarized as follows; $HSP90{\alpha}$ showed higher expression in patients with no lymphovascular invasion (p=0.014). $HSP90{\beta}$ showed a higher expression of squamous cell carcinoma (p=0.003), and an over expression of glucose-related protein (GRP94) was significantly associated with poor differentiation (p=0.048). However, none of the HSP90 proteins showed a significant association with the survival status in patients with NSCLC. This study also indicates that $HSP90{\alpha}$ might contribute more to the carcinogenesis of NSCLC than $HSP90{\beta}$, and GRP94 and isoform selectivity should be considered when HSP90 inhibitors are studied or utilized in the treatment of NSCLC.

Effects of Salt and Drought Stresses on Seed Germination and Gene Expression Pattern in Tall Fescue (염과 건조 스트레스 조건에서 톨 페스큐의 종자 발아율과 유전자 발현 변화분석)

  • Lee, Sang-Hoon;Lee, Ki-Won;Choi, Gi Jun;Kim, Ki-Yong;Ji, Hee Jung;Hwang, Tae Young;Lee, Dong-Gi
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.34 no.2
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    • pp.114-119
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    • 2014
  • Salinity and drought stresses are probably the most significant abiotic factor limiting plant's growth, also negatively affect seed germination and early seedling development. To study on effect of NaCl and PEG stress on seed germination and gene expression pattern of tall fescue, the levels of NaCl and PEG-induced water stresses were determined in first experiment. Different concentration of NaCl (0 to 350 mM) and PEG (0 to 30%) were used for seed treatment. Seed Germination percentage reduced with increasing osmotic potential of growth medium either due to NaCl or PEG. Seeds were not germinate at 350 mM NaCl or 30% PEG treatment. On the basis of the results, Kentucky31(E-) had more resistant than Fawn in both stress conditions. Furthermore, we have used an annealing control primer-based differential display reverse transcription-polymerase chain reaction method to identify salt- and drought stress-induced differentially expressed genes (DEGs) in tall fescue leaves. Using 120 annealing control primers, a total of 4 genes were identified and sequenced. The possible roles of the identified DEGs are discussed in the context of their putative role during salinity and drought stresses.

Dynamic DNA Methylation Change of Dnmt1o 5'-Terminal Region during Preimplantation Development of Cloned Pig (돼지 체세포 복제란 초기발달 과정 중 Dnmt1o 상류 영역의 다이내믹한 DNA 메틸화 변화)

  • Ko, Yeoung-Gyu;Kim, Sung-Woo;Cho, Sang-Rae;Do, Yoon-Jung;Kim, Jae-Hwan;Kim, Sang-Woo;Kim, Hyun;Park, Jae-Hong;Park, Soo-Bong
    • Reproductive and Developmental Biology
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    • v.36 no.1
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    • pp.7-12
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    • 2012
  • DNA methyltransferase 1 (Dnmt1) gene contains three different isoform transcripts, Dnmt1s, Dnmt1o, and Dnmt1p, are produced by alternative usage of multiple first exons. Dnmt1o is specific to oocytes and preimplantation embryos, whereas Dnmt1s is expressed in somatic cells. Here we determined that porcine Dnmt1o gene had differentially methylated regions (DMRs) in 5'-flanking region, while those were not found in the Dnmt1s promoter region. The methylation patterns of the porcine Dnmt1o/Dnmt1s DMRs were investigated using bisulfite sequencing and pyrosequencing analysis through all preimplantation stages from one cell to blastocyst stage in in vivo or somatic cell nuclear transfer (SCNT). The Dnmt1o DMRs contained 8 CpG sites, which located in -640 bp to -30 bp upstream region from transcription start site of the Dnmt1o gene. The methylation status of 5 CpGs within the Dnmt1o DMRs were distinctively different at each stage from one-cell to blastocyst stage in the $in$ $vivo$ or SCNT, respectively. 55.62% methylation degree of the Dnmt1o DMRs in the $in$ $vivo$ was increased up to 84.38% in the SCNT embryo, moreover, $de$ $novo$ methylation and demethylation occurred during development of porcine embryos from the one-cell stage to the blastocyst stage. However, the DNA methylation states at CpG sites in the Dnmt1s promoter regions were hypomethylated, and dramatically not changed through one-cell to blastocyst stage in the $in$ $vivo$ or SCNT embryos. In the present study, we demonstrated that the DMRs in the promoter region of the porcine Dnmt1o was well conserved, contributing to establishment and maintenance of genome-wide patterns of DNA methylation in early embryonic development.

Role of Lysyl Oxidase Family during Odontoblastic Differentiation of Human Dental Pulp Cells Induced with Odontogenic Supplement (인간치수세포에서 상아모세포의 분화과정 동안 Lysyl Oxidase Family의 역할)

  • Lee, Hwa-Jeong;Han, Soo-Yeon
    • Journal of dental hygiene science
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    • v.13 no.3
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    • pp.296-303
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    • 2013
  • Lysyl oxidase (LOX), extracellular matrix enzyme, is catalyzing lysine-derived crosslinks in collagen and elastin. Recently, several LOX-like proteins (LOXL, LOXL2, LOXL3 and LOXL4) have been identified in human but their specific functions are still largely unknown. The purpose of this study was to evaluate the function of the LOX family genes during odontoblastic differentiation of human dental pulp (HDP) cells induced with odontogenic supplement (OS). The messenger RNA (mRNA) expression of LOX family genes and differentiation markers was assessed by reverse transcriptase polymerase chain reaction analysis (RT-PCR). The formation of mineralization nodules was evaluated by alrizarin red S staining. Amine oxidase activity of HDP cells was measured by peroxidase-coupled fluormetric assay. The expressions of differentiation markers, such as alkaline phosphatase (ALP), osteopontin (OPN), osteocalcin (OCN), dentin matrix protein1 (DMP1), dentin sialophosphoprotein (DSPP) in HDP cells were increased after treatment with OS media. The LOX and LOXL mRNA expression were gradually increased in OS media, whereas LOX enzyme activities were markedly detected on day 7. The mRNA expression and LOX enzyme activity of collagen type I was very similar to the pattern of LOX gene. In this study, the expression of LOX and its isoforms, and activity of LOX were highly regulated during odontoblastic differentiation. Thus, these results suggest that LOX plays a key role in odontoblastic differentiation of HDP cells.

Effects of Relative Lysyl Oxidase and Hydrogen Peroxide on Odontoblastic Differentiation (인간치수세포 분화과정에서 과산화수소에 대한 Lysyl Oxidase의 역할)

  • Lee, Hwa-Jeong
    • Journal of dental hygiene science
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    • v.13 no.3
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    • pp.321-329
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    • 2013
  • Although it has been reported that lysyl oxidase (LOX) is involved in odontoblastic differentiation, the role of LOX on odontoblastic differentiation by hydrogen peroxide ($H_2O_2$) have not been clarified. In the present study, we investigated whether $H_2O_2$, reactive oxygen species (ROS), is modulated the messenger RNA (mRNA) expression and activity of LOX during odontoblastic differentiation of human dental pulp (HDP) cells. The mRNA expression was quantified by reverse transcriptase polymerase chain reaction (RT-PCR) analysis, and LOX enzyme activity was measured by high sensitive fluorescent assay. Expression of the odontoblastic differentiation marker genes were assessed in the presence and absence of specific small interfering RNAs (siRNAs) of the LOX and LOXL. The $H_2O_2$-induced mRNA expression of LOX family was significant reduction of LOX, LOXL, and LOXL3 mRNA levels in HDP cells. LOX enzyme activity was increased at $H_2O_2$ 0.3 mM for 24 hours. The mRNA expression of alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN) was inhibited by LOX- and LOXL-specific siRNAs whereas the mRNA expression of dentin matrix protein1 (DMP1), and dentin sialophosphoprotein (DSPP) was inhibited by LOX-specific siRNA. In LOX enzyme activity, siRNA-induced knockdown of both LOX and LOXL inhibited the total amine oxidase activity in HDP cells, as in the case of mRNA expression. In conclusion, the essential role of $H_2O_2$ on odontoblastic differentiation suggests that its regulation by LOX may have pharmacologic importance in HDP cells.