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Hypoxia Inducible Factor-1α Directly Regulates Nuclear Clusterin Transcription by Interacting with Hypoxia Response Elements in the Clusterin Promoter

  • Park, Jeongsook;Park, So Yun;Shin, Eunkyung;Lee, Sun Hee;Kim, Yoon Sook;Lee, Dong Hoon;Roh, Gu Seob;Kim, Hyun Joon;Kang, Sang Soo;Cho, Gyeong Jae;Jeong, Bo-Young;Kim, Hwajin;Choi, Wan Sung
    • Molecules and Cells
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    • v.37 no.2
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    • pp.178-186
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    • 2014
  • Differential transcription of the clusterin (CLU) gene yields two CLU isoforms, a nuclear form (nCLU) and a secretory form (sCLU), which play crucial roles in prostate tumorigenesis. Pro-apoptotic nCLU and anti-apoptotic sCLU have opposite effects and are differentially expressed in normal and cancer cells; however, their regulatory mechanisms at the transcriptional level are not yet known. Here, we examined the transcriptional regulation of nCLU in response to hypoxia. We identified three putative hypoxia response elements (HREs) in the human CLU promoter between positions -806 and +51 bp. Using a luciferase reporter, electrophoretic gel mobility shift, and chromatin immunoprecipitation assays, we further showed that hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) bound directly to these sites and activated transcription. Exposure to the hypoxia-mimetic compound $CoCl_2$, incubation under 1% $O_2$ conditions, or overexpression of HIF-$1{\alpha}$ enhanced nCLU expression and induced apoptosis in human prostate cancer PC3M cells. However, LNCaP prostate cancer cells were resistant to hypoxia-induced cell death. Methylation-specific PCR analysis revealed that the CLU promoter in PC3M cells was not methylated; in contrast, the CLU promoter in LNCap cells was methylated. Co-treatment of LNCaP cells with $CoCl_2$ and a demethylating agent promoted apoptotic cell death through the induction of nCLU. We conclude that nCLU expression is regulated by direct binding of HIF-$1{\alpha}$ to HRE sites and is epigenetically controlled by methylation of its promoter region.

Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli (대장균이 생산한 재조합 인체 감마인터페론의 발현과 정제)

  • Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
    • KSBB Journal
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    • v.21 no.3
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    • pp.204-211
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    • 2006
  • For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

The Concentration of Economic Power in Korea (경제력집중(經濟力集中) : 기본시각(基本視角)과 정책방향(政策方向))

  • Lee, Kyu-uck
    • KDI Journal of Economic Policy
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    • v.12 no.1
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    • pp.31-68
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    • 1990
  • The concentration of economic power takes the form of one or a few firms controlling a substantial portion of the economic resources and means in a certain economic area. At the same time, to the extent that these firms are owned by a few individuals, resource allocation can be manipulated by them rather than by the impersonal market mechanism. This will impair allocative efficiency, run counter to a decentralized market system and hamper the equitable distribution of wealth. Viewed from the historical evolution of Western capitalism in general, the concentration of economic power is a paradox in that it is a product of the free market system itself. The economic principle of natural discrimination works so that a few big firms preempt scarce resources and market opportunities. Prominent historical examples include trusts in America, Konzern in Germany and Zaibatsu in Japan in the early twentieth century. In other words, the concentration of economic power is the outcome as well as the antithesis of free competition. As long as judgment of the economic system at large depends upon the value systems of individuals, therefore, the issue of how to evaluate the concentration of economic power will inevitably be tinged with ideology. We have witnessed several different approaches to this problem such as communism, fascism and revised capitalism, and the last one seems to be the only surviving alternative. The concentration of economic power in Korea can be summarily represented by the "jaebol," namely, the conglomerate business group, the majority of whose member firms are monopolistic or oligopolistic in their respective markets and are owned by particular individuals. The jaebol has many dimensions in its size, but to sketch its magnitude, the share of the jaebol in the manufacturing sector reached 37.3% in shipment and 17.6% in employment as of 1989. The concentration of economic power can be ascribed to a number of causes. In the early stages of economic development, when the market system is immature, entrepreneurship must fill the gap inherent in the market in addition to performing its customary managerial function. Entrepreneurship of this sort is a scarce resource and becomes even more valuable as the target rate of economic growth gets higher. Entrepreneurship can neither be readily obtained in the market nor exhausted despite repeated use. Because of these peculiarities, economic power is bound to be concentrated in the hands of a few entrepreneurs and their business groups. It goes without saying, however, that the issue of whether the full exercise of money-making entrepreneurship is compatible with social mores is a different matter entirely. The rapidity of the concentration of economic power can also be traced to the diversification of business groups. The transplantation of advanced technology oriented toward mass production tends to saturate the small domestic market quite early and allows a firm to expand into new markets by making use of excess capacity and of monopoly profits. One of the reasons why the jaebol issue has become so acute in Korea lies in the nature of the government-business relationship. The Korean government has set economic development as its foremost national goal and, since then, has intervened profoundly in the private sector. Since most strategic industries promoted by the government required a huge capacity in technology, capital and manpower, big firms were favored over smaller firms, and the benefits of industrial policy naturally accrued to large business groups. The concentration of economic power which occured along the way was, therefore, not necessarily a product of the market system. At the same time, the concentration of ownership in business groups has been left largely intact as they have customarily met capital requirements by means of debt. The real advantage enjoyed by large business groups lies in synergy due to multiplant and multiproduct production. Even these effects, however, cannot always be considered socially optimal, as they offer disadvantages to other independent firms-for example, by foreclosing their markets. Moreover their fictitious or artificial advantages only aggravate the popular perception that most business groups have accumulated their wealth at the expense of the general public and under the behest of the government. Since Korea stands now at the threshold of establishing a full-fledged market economy along with political democracy, the phenomenon called the concentration of economic power must be correctly understood and the roles of business groups must be accordingly redefined. In doing so, we would do better to take a closer look at Japan which has experienced a demise of family-controlled Zaibatsu and a success with business groups(Kigyoshudan) whose ownership is dispersed among many firms and ultimately among the general public. The Japanese case cannot be an ideal model, but at least it gives us a good point of departure in that the issue of ownership is at the heart of the matter. In setting the basic direction of public policy aimed at controlling the concentration of economic power, one must harmonize efficiency and equity. Firm size in itself is not a problem, if it is dictated by efficiency considerations and if the firm behaves competitively in the market. As long as entrepreneurship is required for continuous economic growth and there is a discrepancy in entrepreneurial capacity among individuals, a concentration of economic power is bound to take place to some degree. Hence, the most effective way of reducing the inefficiency of business groups may be to impose competitive pressure on their activities. Concurrently, unless the concentration of ownership in business groups is scaled down, the seed of social discontent will still remain. Nevertheless, the dispersion of ownership requires a number of preconditions and, consequently, we must make consistent, long-term efforts on many fronts. We can suggest a long list of policy measures specifically designed to control the concentration of economic power. Whatever the policy may be, however, its intended effects will not be fully realized unless business groups abide by the moral code expected of socially responsible entrepreneurs. This is especially true, since the root of the problem of the excessive concentration of economic power lies outside the issue of efficiency, in problems concerning distribution, equity, and social justice.

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Fractionation and Availability of Heavy Metals in Paddy Soils near Abandoned Mining Areas (광산인근 논토양의 중금속 분획화 및 유효도)

  • Jung, Goo-Bok;Kim, Won-Il;Ryu, In-Soo
    • Korean Journal of Environmental Agriculture
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    • v.19 no.4
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    • pp.319-323
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    • 2000
  • This study was conducted to compare fractionations and availability of heavy metal in paddy soils near five abandoned mining areas. The sequential extraction procedure was used to fractionate the heavy metals in soils into the designated from water $soluble(H_2O)$, $exchangeable(0.5M\;KNO_3)$, organically bound(0.5M NaOH), $oxide/carbonate(0.05M\;Na_2-EDTA)$, and $sulfide/residual(4M\;HNO_3)$. EDTA and $HNO_3$ extractable of Cd, Pb, and Zn, and NaOH and $HNO_3$, extractable of Cu were predominant chemical forms. The ratio of $H_2O+KNO_3$ extractable of Cd, Zn, Cu, and Pb were 25.1, 8.7, 4.0, and 0.4%, respectively. The ratio of $H_2O+KNO_3$ extractable heavy metal were negatively correlated with soil pH, while $EDTA+HNO_3$ extractable heavy metal were positively correlated. The most consistent distribution patterns were found when the soil samples were grouped according to their total contents. Specially, the ratio of $H_2O+KNO_3$ extractable heavy metal were higher as total contents of heavy metal were increased. The ratio of $H_2O+KNO_3$ extractable heavy metal(Cd 1.06, Cu 0.15, Pb 0.01, and Zn 0.05%) were lower at the high soil pH than those(Cd 31.31, Cu 4.06, Pb 1.75, and Zn 10.16%) at the low level. Compared to other chemical forms, the degree of contribution for $KNO_3$ extractable form to the Cd uptake to brown rice was high, whereas that for EDTA and $HNO_3$ extractable forms were high to the Zn.

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Empty Time, Empty Space, and Non(非)-Place in World of Warcraft: The Accumulated Experience and the Recovery in Reality (<월드 오브 워크레프트>(World of Warcraft)의 빈 시간, 빈 공간, 비(非) 장소: 축적된 체험과 실감의 회복)

  • Song, Kyong-Won
    • Cartoon and Animation Studies
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    • s.19
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    • pp.127-143
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    • 2010
  • The purpose of this study is to aesthetically approach a game, which was popular at home, through microscopic analysis on World of Warcraft. World of Warcraft(hereunder called WOW), which improved a game method of MMORPG that had been popular conventionally at home, from several angles, is suggesting new paradigm of MMORPG game. This study aims to propose new vision on game analysis by approaching this new method from the perspective of player. For this, it divided WOW into shift method, instruction implementation method, and communication configuration method, and analyzed this, respectively, by introducing a concept of empty time, empty space, and non-place, which was borrowed from Zygmunt Bauman's "Liquid Modernity". WOW's shifting method rather extends empty time, that is, the waiting time, contrary to the conventional MMORPG, which used a method of removing. Instead of maximally reducing boring, which becomes disturbance of a game, it is what overwhelmingly enlarges time that needs to be waited, thereby being what induces a sense of time, which a player experiences daily life, to the inside of a game. WOW's instruction implementation method offers one of hugely single map instead of zone-system necessary for map loading in the discontinued form. This minutely implements even a greatly insignificant place in playing a game, thereby stirring up a sense of travel, which explores there in reality. Finally, WOW's community configuration method makes a hunting-targeted group clear, which added a concept of hunting called the raid to the existing gild system. The raid, which is a large party of being bound to the inside of Instant Dungeon, clearly gives a performance role to each of party members, thereby allowing the identity in character to be connected directly to the identity of player. Through this, the player filled the suggested 'space' with experience, thereby being able to change it into 'place' that is significant to an individual.

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The Study of Satire Shown in Animation -Focusing on and (애니메이션에 나타난 풍자성 연구 -<대화의 차원>과 <이웃>을 중심으로)

  • Choi, Don-Ill
    • Cartoon and Animation Studies
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    • s.44
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    • pp.143-161
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    • 2016
  • This study was conducted focusing on the role of auteurism animation. The purpose of auteurism animation is to criticize irregularities of a society in witty and fierce way through satires from the sharp perspectives of a animator that is not bound by tastes of people or the interests or standpoints of specific groups, and thus to induce positive changes in a society as a purifier. In the context, this study investigated satires shown in by Jan Svankmajer and by Norman Mclaren among the animators who utilize animation as a tool to produce social meaning. As a result, the following characteristics and meanings were found. First, Dimensions of Dialogue is an animation that satires absurdity and irregularities of a human society in symbolic and exceptional way through directing by segmentations of images and omnibus structures. The satire carries the lesson of improvement in the hidden part of cynical attack to history, society, and human beings. It also maximizes absolute reality and engagement of images of Jan Svankmajer through unique and grotesque images of the animator such as alienated world, confusing shapes, and amusement of irregularities. Second, the movie, is an exemplary animation that applied core concept of animation through pixilation techniques based on an event story structure by causal relationship. It satires the changing process of a good man to violent madness through confrontation and conflicts for material desires, with exaggerated slipstick movements and humors as a black comedy. The satire methods of both animation works are delivered through unique image styles and symbolic wordage of the animators who triggered ironical laughter in attacking humanism and moral insensitivity that might be felt seriously otherwise. That is, the animators try to show the positive will for changing the society to a sound one through the form of negativity in terms of moral perspective in animation rather than destruction against the target. As such, the satires in both works worked as an auteurism allegory that maximizes social functions and artistic influence of animation.

Assessment Techniques of Heavy Metal Bioavailability in Soil - A critical Review (토양 중 중금속 생물유효도 평가방법 - 총설)

  • Kim, Kwon-Rae;Owens, Gary;Naidu, Ravi;Kim, Kye-Hoon
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.4
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    • pp.311-325
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    • 2007
  • The concept of metal bioavailability, rather than total metal in soils, is increasingly becoming important for a thorough understanding of risk assessment and remediation. This is because bioavailable metals generally represented by the labile or soluble metal components existing as either free ions or soluble complexed ions are likely to be accessible to receptor organismsrather than heavy metals tightly bound on soil surface. Consequently, many researchers have investigated the bioavailability of metals in both soil and solution phases together with the key soil properties influencing bioavailability. In order to study bioavailability changes various techniques have been developed including chemical based extraction (weak salt solution extraction, chelate extraction, etc.) and speciation of metals using devices such as ion selective electrode (ISE) and diffusive gradient in the thin film (DGT). Changes in soil metal bioavailability typically occur through adsorption/desorption reactions of metal ions exchanged between soil solution and soil binding sites in response to changes in environment factors such as soil pH, organic matter (OM), dissolved organic carbon (DOC), low-molecular weight organic acids (LMWOAs), and index cations. Increasesin soil pH result in decreases in metal bioavailability through adsorption of metal ions on deprotonated binding sites. Organic matter may also decrease metal bioavailability by providing more negatively charged binding sites, and metal bioavailability can also be decreases as concentrations of DOC and LMWOAs increase as these both form strong chelate complexeswith metal ions in soil solution. The interaction of metal ions with these soil properties also varies depending on the soil and metal type.

Influence of Phenobarbital on the Circadian Rhythm of Opiate Receptor in Rat Brain (백서의 뇌내 Opiate 수용체의 일중 변동에 미치는 Phenobarbital의 영향)

  • Park, Yeoung-Gul;Kim, Kee-Won;Cho, Kyu-Park
    • The Korean Journal of Pharmacology
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    • v.21 no.2
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    • pp.128-141
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    • 1985
  • To investigate the influence of phenobarbital sodium on the action of morphine and on the diurnal rhythms of both opiate receptor binding and ${\beta}-endorphin$ contents, the amount of specifically bound $(^3H)$-morphine and immunoreactive ${\beta}-endorphin$ were measured in the midbrain of phenobarbital-treated rats at 4h intervals in a day. Rats were housed and adapted to a controlled cycle of either 12 h light-12 h dark or 24 h constant dark. After 3 weeks of adaptation, 0.5 ml of physiological saline or phenobarbital sodium (20mg/kg/day, i.p.) were administered twice a day for 2 weeks. Highly significant diurnal rhythms of opiate receptor binding and ${\beta}-endorphin$ were present in rat midbrain. In control group, the peak of maximum $(^3H)$-morphine binding was observed at 22:00 h, whereas the peak of ${\beta}-endorphin$ content was found at 06:00 h. Even in the absence of time cues these diurnal rhythms persisted, but they were highly modified with respect to the wave form as well as differences in the timing of peak and nadir. In the phenobarbital-treated group, these diurnal rhythms were also modified in shape, phase and amplitude, as well as in timing of peak and nadir. In this group, 24 h mean of opiate receptor binding was significantly decreased, while the 24 h mean level of ${\beta}-endorphin$ content was highly increased. However, Kd values in all experimental groups did not change. This indicates that differences in binding were not due to changes in the affinity, but in the number of binding sites. Statistical analysis of regression line indicates that changes of receptor binding were closely correlated with the changes of ${\beta}-endorphin$ content. These results suggest that phenobarbital may influence the action of morphine by changing the number of opiate receptors and that the modification of diurnal rhythm of opiate receptor by the agent is possibly due to changes of ${\beta}-endorphin$ content.

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Autoradiographic Studies on the Inhibitory Effect of Dibutyryl Cyclic AMP on Mouse Oocyte Maturation in Vitro (Dibutyryl Cyclic AMP가 생쥐여포난자의 성숙에 미치는 억제효과에 관한 자기방사법적 연구)

  • Choi, Choon-Keun
    • Applied Microscopy
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    • v.7 no.1
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    • pp.21-43
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    • 1977
  • This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

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sICAM-1, sE-selectin, sVCAM-1 Concentration in Patients with Pulmonary Tuberculosis (폐결핵 환자에서 SICAM-1, sE-selectin sVCAM-1농도의 변화)

  • Oh, Sang-Mi;Jang, Jae-Ho;Choi, Sang-In;Lee, Heung-Bum;Lee, Yong-Chul;Rhee, Yang-Keun
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1256-1262
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    • 1997
  • Background : Leukocyte-endothelial adhesion molecules have been implicated in the pathogenesis of inflammatory disease. ICAM-1, VCAM-1 and E-selectin are cell surface adhesion molecule on vascular endothelial cells. They are up-regulated by inflammatory cytokines and regulate the adhesion and migration of leukocytes across the endothelium. Tuberculosis, a granulomatous disorder is an infection caused by Mycobacterium tuberculosis. The clinical manifestations of tuberculosis are dependent on the cellular immune response to tubercule bacilli. Circulating adhesion molecules are probably formed by cleavage and release into the circulation of the extracellular domain of the membrane bound form. The elevated levels of circulating adhesion molecules have been reported in numerous disease state. To evaluate their role as markers of disease activity in tuberculosis, we measured a sE-selectin, sVCAM-1 and sICAM-1 levels in the serum with severities of mild, moderate and far advanced pulmonary tuberculosis. Methods : The control and test groups were divided as follows. Group I : control(n=5), Group II : patients with mild pulmonary tuberculosis(n=12), Group III : pateints with moderate pulmonary tuberculosis(n=20), Group IV : patients with far advanced pulmonary tuberculosis(n=19). Serum sICAM-1, sVCAM-1 and sE-selectin were measured by ELISA kit Results : Serum soluble adhesion molecules are elevated in patients with pulmonary tuberculosis, Circulating ICAM-1 levels were significantly elevated in patients with moderate and far advanced pulmonary tuberculosis when compared with control group. When compared with control group, serum sVCAM-1 levels showed significant elevation in patients with mild, moderate and far advanced pulmonary tuberculosis. Serum sE-selectin levels were significantly elevated in patients with far advanced pulmonary tuberculosis when compared with control group. Conclusion : These results suggest that sICAM-1, sVCAM-1, and sE-selectin may be invloved in the pathogenesis of tuberculosis. And, particularly, sICAM-1 and sVCAM-1 may be useful markers of the disease activity.

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