• Title/Summary/Keyword: Direct Expression

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Endothelial Cell Products as a Key Player in Hypoxia-Induced Nerve Cell Injury after Stroke

  • Cho, Chul-Min;Ha, Se-Un;Bae, Hae-Rahn;Huh, Jae-Taeck
    • Journal of Korean Neurosurgical Society
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    • v.40 no.2
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    • pp.103-109
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    • 2006
  • Objective : Activated endothelial cells mediate the cascade of reactions in response to hypoxia for adaptation to the stress. It has been suggested that hypoxia, by itself, without reperfusion, can activate the endothelial cells and initiate complex responses. In this study, we investigated whether hypoxia-induced endothelial products alter the endothelial permeability and have a direct cytotoxic effect on nerve cells. Methods : Hypoxic condition of primary human umbilical vein endothelial cells[HUVEC] was induced by $CoCl_2$ treatment in culture medium. Cell growth was evaluated by 3,4,5-dimethyl thiazole-3,5-diphenyl tetrazolium bromide [MTT] assay Hypoxia-induced products [$IL-1{\beta},\;TGF-{\beta}1,\;IFN-{\gamma},\;TNF-{\alpha}$, IL-10, IL-6, IL-8, MCP-l and VEGF] were assessed by enzyme-linked immunosorbent assay. Endothelial permeability was evaluated by Western blotting. Results : Prolonged hypoxia caused endothelial cells to secrete IL -6, IL -8, MCP-1 and VEGF. However, the levels of IL -1, IL -10, $TNF-{\alpha},\;TGF-{\beta},\;IFN-{\gamma}$ and nitric oxide remained unchanged over 48 h hypoxia. Hypoxic exposure to endothelial cells induced the time-dependent down regulation of the expression of cadherin and catenin protein. The conditioned medium taken from hypoxic HUVECs had the cytotoxic effect selectively on neuroblastoma cells, but not on astroglioma cells. Conclusion : These results suggest the possibility that endothelial cell derived cytokines or other secreted products with the increased endothelial permeability might directly contribute to nerve cell injury followed by hypoxia.

Rhamnose-rich and fucose-rich oligo- and polysaccharides (RROP-s and FROPs), agonists and antagonists of cell-membrane receptors as new active principles against skin aging.

  • Robert, L.;Robert, A.M.;Gesztes, J.L.;Luppi, E.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.352-373
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    • 2003
  • Rhamnose-rich (RROP-s) and fucose-rich (FROP-s) oligo-and polysaccharides were prepared and extensively characterised by physical and chemical procedures [1,2] and compared to L-fucose. Their biological properties were then studied on human skin fibroblast cell cultures, human skin explant cultures and on hairless rat skin, using a variety of cell-biological, biochemical and computerised morphometrical procedures. Among the most important properties we could establish, the following are of particular interest for the tretment and prevention of age-dependent modifications of human skin (loss of skin-tissue, cells and matrix, wrinkle formation and others) : stimulation of cell proliferation (by $^3$[H]-thymidine incorporation and the MTT test), scavenging of reactive oxygen species (ROS) using several different procedures, and protease (MMP-2 and MMP-9) down-regulation. A topical preparation, using RROP-s and FROP-s, and/or L-fucose, was shown to increase cell proliferation, dermal matrix synthesis, efficient scavenging of ROS-s and to increase also the thickness of dermal tissue when applied for 4 weeks on hairless rat skin, accompanied by the densification of collagen bundles as well as by an increase of elastin synthesis. Using fluorescent labeled FROPs, it could be shown that these oligosaccharides react with cell-membrane receptors and especially with the elastin-laminin-receptor and the fucose-mannose receptor, but they penetrate also in the cell nucleus, suggesting the possibility of a direct action on the regulation of gene expression. When applied to the human skin of a team of voluntary women encompassing all age-groups, the efficiency of FROP-containing preparation could be confirmed using indentometry and computerised evaluation of skin micro-relief, as well as evaluation of periorbital wrinkles. It appears therefore that these preparations correspond to all the requirements of active anti-aging principles.

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Effect of DHEA on Recovery of Muscle Atrophy Induced by Parkinson' s Disease

  • Choe, Myoung-Ae;An, Gyeong-Ju;Koo, Byung-Soo;Jeon, Song-Hee
    • Journal of Korean Academy of Nursing
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    • v.41 no.6
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    • pp.834-842
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    • 2011
  • Purpose: The purpose of this study was to determine the effect of dehydroepiandrosterone (DHEA) on recovery of muscle atrophy induced by Parkinson's disease. Methods: The rat model was established by direct injection of 6-hydroxydopamine (6-OHDA, 20 ${\mu}g$) into the left striatum using stereotaxic surgery. Rats were divided into two groups; the Parkinson's disease group with vehicle treatment (Vehicle; n=12) or DHEA treatment group (DHEA; n=22). DHEA or vehicle was administrated intraperitoneally daily at a dose of 0.34 mmol/kg for 21 days. At 22-days after DHEA treatment, soleus, plantaris, and striatum were dissected. Results: The DHEA group showed significant increase (p<.01) in the number of tyrosine hydroxylase (TH) positive neurons in the lesioned side substantia nigra compared to the vehicle group. Weights and Type I fiber cross-sectional areas of the contralateral soleus of the DHEA group were significantly greater than those of the vehicle group (p=.02, p=.00). Moreover, extracellular signal-regulated kinase (ERK) phosphorylation significantly decreased in the lesioned striatum, but was recovered with DHEA and also in the contralateral soleus muscle, Akt and ERK phosphorylation recovered significantly and the expression level of myosin heavy chain also recovered by DHEA treatment. Conclusion: Our results suggest that DHEA treatment recovers Parkinson's disease induced contralateral soleus muscle atrophy through Akt and ERK phosphorylation.

THE EFFECTS OF SODIUM FLUORIDE ON TYPE I $\alpha$ 2 COLLAGEN RIBONUCLEIC ACID (mRNA) LEVEL IN MURIN OSTEOBLAST LIKE (MC3T3-E1) CELLS (Sodium Fluoride가 조골세포주 MC3T3-E1의 제 1 형 ${\alpha}2$ 교원질 mRNA에 미치는 영향에 관한 연구)

  • Hwang, Jeung-Bin;Chung, Kyu-Rhim;Park, Young-Guk
    • The korean journal of orthodontics
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    • v.23 no.3 s.42
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    • pp.415-425
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    • 1993
  • Fluoride is one of the most potent stimulators of bone formation in vivo. But its direct effects on osteoblast is not yet clear This study was to investigate the effects of Sodium fluoride on alkaline phosphatase(ALP) activity, cAMP formation responsive to parathormone(PTH) and type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level in Murin osteoblast-like (MC3T3-E1) cells. The cells were cultured in $\alpha-Minimal$ essential medium $(\alpha-MEM)$ supplemente with $10\%$ fetal bovine serum (FBS) and then changed to $0.1\%$ FBS with various concentration of Sodium fluoride. The ALP activity was assayed by the method of Lowry with disodium phenyl phosphated as substrate. cAMP formation was measured by Radioimmuno Assay(RIA). Type I $\alpha$ 2 collagen ribonucleic acid(mRNA) expression was studied by Nothern blot analysis. The results were as follows: 1. cAMP level was increased by PTH in MC3T3-E1 cells. 2. Sodium fluoride showed the tendency of inhibitory effects on cAMP responsiveness to PTH in MC3T3-E1 cells. 3. Sodium fluoride increased ALP activity at cocentration of $2{\mu}M,\;4{\mu}M,\;and\;10{\mu}M$ significantly different from control at the 0.001 level. ALP activity revealed maximum value at $10{\mu}M$ in this study. 4. Nothern blot analysis of Sodium fluoride treated cells, using Type I $\alpha$ 2 collagen prove, revealed significant increase at $10{\mu}M$ in MC3T3-E1 cells.

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Effect of Mineral Trioxide Aggregate and Calcium Hydroxide on Reparative Dentin Formation in Rats

  • Ra, Ji-Young;Lee, Wan;Kim, Hyun-Jin
    • International Journal of Oral Biology
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    • v.37 no.2
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    • pp.77-83
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    • 2012
  • We investigated the pulpal response to direct pulp capping in rat molar teeth using mineral trioxide aggregate (MTA) and calcium hydroxide (CH). A palatal cavity was prepared in rat maxillary molar teeth. Either MTA or CH was placed on the exposed pulp and all cavities were restored with composite. Rats were sacrificed for histological evaluation after 12 hours and at 2, 7, 14 and 21 days. In both the MTA and CH groups, reparative dentin formation was clearly observed on histology after 14 days. The MTA-capped pulps were found to be mostly free from inflammation, and hard tissue of a tubular consistent barrier was observed. In contrast, in CH-capped teeth, excessive formation of reparative dentin toward residual pulp was evident. The pulpal cell response beneath the reparative dentin layer was examined by immunofluorescence using antibodies against DSP. After 2 days, a few DSP immunopositive cells, most of which showed a cuboidal shape, appeared beneath the predentin layer. At 7 days, DSP-immunopositive cells with columnar odontoblast-like cells were seen beneath the newly formed hard tissues. At 14 and 21 days, DSP was more abundant in the vicinity of the odontoblastic process along the dentinal tubules than in the mineralized reparative dentin. The CH group showed strong expression patterns in terms of DSP immunoreactivity. Our results thus indicate that MTA may be a more effective pulp capping material as it induces the differentiation of odontoblast-like cells and the formation of reparative dentin without the loss of residual pulp functions.

In situ Recovery of hCTLA4Ig from Suspension Cell Cultures of Oryza sativa (형질전환 벼 현탁세포 배양에서 hCTLA4Ig의 in situ 회수)

  • Choi, Hong-Yeol;Cheon, Su-Hwan;Kwon, Jun-Young;Yun, Boreum;Hong, Seok-Mi;Kim, Sun-Dal;Kim, Dong-Il
    • KSBB Journal
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    • v.31 no.4
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    • pp.284-290
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    • 2016
  • In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

회원 신윤복 풍용도에 표현된 복식미의 연구

  • 김인경
    • Journal of the Korean Society of Costume
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    • v.25
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    • pp.5-20
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    • 1995
  • The fashion of late Yi dynasty had explored a possibility of new fashional beauty from its own experiences throughout the history of Korean fashion excluding any interference of foreign fashions, and brought out some characteristics ; first, the aesthetical value focused on the more human fashion resulted in the exposure of waist. This aesthetical change in the fashion of late Yi dynasty produced out results as follows ; the length and width of jacket became short and narrow maintaining the basic form, while the width of skirt became increased to be contrasted exceptionally with the upper coat. This mode of fashion reveals the transitional movement from the emphasis on balance to that on unbalance in the late period of Yi dynasty. But regarding the fact that the proportional change by the ways of putting on skirt became to be similar to the golden ratio, we can see many examples throughout the folk paintings by Shin Yun bok, we find that the fashional beauty of that age pursued ideal harmony and unity all the time being in spite of its tendency toward non-refinement. Second, we see another peculiarity in the mode of woman's fashion from the paintings by Shin Yun-bok who always depicted the hu-man nature frankly. For example, the jacket became shorter to emphasize lady's slim waist, and underwears began to be exposed after the upper part of skirt had been to expose lady's waist while the bottom line pulled up highter than before. Thus, as the function of robe had changed from a mere means of covering into an expression of beauty, the fashion of late Yi dynasty began to express a perfectly new beauty possessing an erotic mood and sensu-ality of a woman, which was its inner aesthetic need. Third, this emergence of new fashional beauty made the mental value of man and the aesthetic views of that time quite different from the previous ones, and finally there came various expressions of a new beauty. There-fore, the woman's fashion of late Yi dynasty is noted for the exposure and emphasis on the body line of a woman after the aesthetic view-point had changed of time toward pragmatism and humanism, while upholding the traditions of Korean clothes and pride for the nation. And our ancestors realized very Korean fashional beauty by pursuing the expressional ways of gentle, indirect emphasis upon the feminine and natural beauty to exclude and ar-tificial, direct and straightforward exposures and images.

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Drug resistance of bladder cancer cells through activation of ABCG2 by FOXM1

  • Roh, Yun-Gil;Mun, Mi-Hye;Jeong, Mi-So;Kim, Won-Tae;Lee, Se-Ra;Chung, Jin-Woong;Kim, Seung Il;Kim, Tae Nam;Nam, Jong Kil;Leem, Sun-Hee
    • BMB Reports
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    • v.51 no.2
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    • pp.98-103
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    • 2018
  • Recurrence is a serious problem in patients with bladder cancer. The hypothesis for recurrence was that the proliferation of drug-resistant cells was reported, and this study focused on drug resistance due to drug efflux. Previous studies have identified FOXM1 as the key gene for recurrence. We found that FOXM1 inhibition decreased drug efflux activity and increased sensitivity to Doxorubicin. Therefore, we examined whether the expression of ABC transporter gene related to drug efflux is regulated by FOXM1. As a result, ABCG2, one of the genes involved in drug efflux, has been identified as a new target for FOXM1. We also demonstrated direct transcriptional regulation of ABCG2 by FOXM1 using ChIP assay. Consequently, in the presence of the drug, FOXM1 is proposed to directly activate ABCG2 to increase the drug efflux activation and drug resistance, thereby involving chemoresistance of bladder cancer cells. Therefore, we suggest that FOXM1 and ABCG2 may be useful targets and important parameters in the treatment of bladder cancer.

A Case of Conversion Disorder Treated with Gaegyeolseogyeong-tang (전환장애 환자에 개결서경탕(開結舒經湯)을 투여한 치험례)

  • Yoon, Ji-Won;Kim, Hong-Joon;Kim, Woo-Sung;Sim, Kuk-Jin;Shim, Ha-Na;Lee, Sang-Kwan;Kang, Sei-Young
    • The Journal of Internal Korean Medicine
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    • v.25 no.3
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    • pp.590-595
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    • 2004
  • Conversion Disorder is a disorder whose predominant feature is a loss or alteration in physical functioning that suggests a physical disorder but that is actually a direct expression of a psychological conflict or need. The Diagnostic and Statistical Manual of Mental Disorders-IV (DSM-IV) guidelines for Conversion Disorder include these definitions: A psychosocial stressor produces a psychological conflict that is believed to help initiate or exacerbate the illness The symptoms are not under conscious control, etc. While functional disabilities are common with conversion disorders, physical and laboratory abnormalities are absent or minor in comparison with the patient's subjective complaints. Symptoms of Conversion Disorder are similar to those of stroke. But the mechanism of Conversion Disorder is similar as that of Stagnation Syndrome of Ki (氣鬱證) in Oriental medicine. Gaegyeolseogyeong-tang has been used to treat women who suffer from Conversion Disorder induced by the Stagnation Syndrome of Ki (氣鬱證). After application of the Gaegyeolseogyeong-tang for 7 days, symptoms and signs improved dramatically.

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Isolation and characterization of a cDNA encoding a mammalian cathepsin L-like cysteine proteinase from Acanthmoeba healui

  • Hong, Yeon-Chul;Hwang, Mi-Yul;Yun, Ho-Cheol;Yu, Hak-Sun;Kong, Hyun-Hee;Yong, Tai-Soon;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.40 no.1
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    • pp.17-24
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    • 2002
  • We have cloned a cDNA encoding a cysteine proteinase of the Acanthamoeba healui OC-3A strain isolated from the brain of a granulomatous amoebic encephalitis patient. A DNA probe for an A. healui cDNA library screening was amplified by PCR using degenerate oligonucleotide primers designed on the basis of conserved amino acids franking the active sites of cysteine and asparagine residues that are conserved in the eukaryotic cysteine proteinases. Cysteine proteinase gene of A. healui (AhCPI) was composed of 330 amino acids with signal sequence, a proposed pro-domain and a predicted active site made up of the catalytic residues, $Cys^{25},{\;}His^{159},{\;}and{\;}Asn^{175}$. Deduced amino acid sequence analysis indicates that AhCPI belong to ERFNIN subfamily of C 1 peptidases. By Northern blot analysis. no direct correlation was observed between AhCPI mRNA expression and virulence of Acanthamoeba, but the gene was expressed at higher level in amoebae isolated from soil than amoeba from clinical samples. These findings raise the possibility that AhCPI protein may play a role in protein metabolism and digestion of phagocytosed bacteria or host tissue debris rather than in invasion of amoebae into host tissue.