• 제목/요약/키워드: Culture medium

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Increased Protein of the Secretory Leukocyte Pretense Inhibitor (SLPI) and the Expression of Growth Factors in NIH3T3 Cells by LPS Stimulation (NIH3T3 세포주에서 LPS자극에 의한 분비백혈구단백분해효소억제제 (SLPI)의 단백질증가와 성장인자들의 발현)

  • Lee, Sang-Hwa;Choi, Baik-Dong;Jeong, Soon-Jeong;Jang, Hyun-Seon;Kim, Byung-Ock;Lim, Do-Seon;Park, Joo-Cheol;Wang, Guan-Lin;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.36 no.3
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    • pp.165-172
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    • 2006
  • Secretory leukocyte protease inhibitor (SLPI) involves tissue protection against the destructive action of neutrophil elastase at the site of inflammation. Several studies on new functions of SLPI have demonstrated that SLPI may play a primary role in innate immunity than protease inhibitor, To identify the function of SLPI by lipopolysaccharide (LPS) stimulation in the embryonic fibroblast (NIH3T3) cells. we studied the expression of SLPI compared to other growth factors involving the LPS treatment. To address this, we performed the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of the SLPI and some growth factors such as VEGF. bFGF, and PDGF-BB after LPS stimulation. NIH3T3 cells were exposed 100 ng/mL Escherichia coli LPS for 30min, 60min, 90min, 24h, and 48h, respectively. The result of RT-PCR showed that SLPI and VEGF mRNA was expressed strongly in NIH3T3 without related to LPS stimulation. mRNA of bFGF was weakly expressed such as the expression of the control. PDGF mRNA expression gradually increased follows at time course. However, SLPI protein level was increased in lysates and culture medium by LPS stimulation. Phase contrast microscopic and scanning electron microscopic observation showed that the increased cell number and cytoplasmic enlargement of the NIH3T3 cells. Therefore, it suggests that the LPS upregulates SLPI expression in NIH3T3 cells. Moreover, secreted SLPI may stimulate cell proliferation and migration.

Serotype and Leukotoxic Strain Distribution of Actinobacillus(Haemophilus) Actinomycetemcomitans in Korean Localized Juvenile Periodontitis (한국인 국소 유년성 치주염환자의 Actinobacillus(Haemophilus) Actinomycetemcomitans 혈청형 및 백혈구독성 균주 분포)

  • Chung, Hyun-Ju;Chung, Chong-Pyoung;Son, Seong-Heui
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.487-501
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    • 1986
  • Previous studies from our laboratory suggested that Korean LJP patients might habor A. actinomycetemcomitans of different serotype from Caucasian LJP patients in whom serotype b was predominant. In order to observe the prevalence and serotype distribution of A. actinomycetemcomitans in localized juvenile periodontitis patients and to evaluate leukotoxic activity of oral isolates, this study was performed. A. actinomycetemcomitans was isolated by using a selective medium(tryptic soy agar supplemented with 10% serum, $75{\mu}g$ of bacitracin and $5{\mu}g$ of vancomycin per ml). Using immunoabsorbed, ammonium sulfate-fractionated serotype-specific antisera, a total of 69 strains were serologically categorized by ELISA. Leukotoxicity was monitored biochemically by measuring lactate dehydrogenase indicator of cell viability in culture supernatant of PMNL plus viable A. actinomycetemcomitans mixture. The results were as follows: 1. A. actinomycetemcomitans was detected in 75% of 16 LJP patients, and 71% in the LJP lesions and 6% in the control sites. 2. Presence or absence of A. actinomycetemcomitans in the sampled disease sites has no in fluence on clinical measurements. 3. Three serotypes were approximately equally distributed in overall 9 patients. Three patients harbored 2 different serotypes of A. actinomycetemcomitans in the same disease site or different disease sites. 4. The proportion of leukotoxic oral isolates was 22% of a total of 46 strains and the prevalence was 69% in 13 sampled sites. The same disease site could harbor both leukotoxic and nonleukotoxic strains. 5. Distribution of leukotoxic strains in 3 serotypes were not different.

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Plant Architecture and Dry Matter Production in Large-Grain Rice Variety (대립벼의 생산구조 특성과 물질생산)

  • 김정곤;전병태;김진기
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.5
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    • pp.655-661
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    • 1995
  • This study was conducted to get basic information of dry matter production and agronomic characteristics for increasing grain ripening in rice cultivar with large grains. Three cultivars which have different grain size were used. Of the two large grain cultivars, SR7796-3-1-5-1-l(SR7796F$_{7}$) had long culm and SR11269-13-3-4-2(SR11269F$_{6}$) had short culm. On the other hand, Tamjinbyeo was recommended cultivar had medium size of grain and culm. LAI was highest in Tamjinbyeo, followed by the SR11269F$_{6}$ and SR7796F$_{7}$, LAI decrement was higher in large grain cultivars than that of Tamjinbyeo after 20days after heading. Total dry weight(TDW) of areal part was highest in SR11269F$_{6}$, followed by the Tamjinbyeo and SR7796F$_{7}$. Dry weight (DW) of stem (leaf sheath+culm) at harvesting stage was decreased 15%, 12% in Tamjinbyeo, SR11269F$_{6}$, respectively, but was increased 5% in SR7796F$_{7}$ compared to heading stage. In distribution rate of each organ to TDW, distribution rate of leaf blade was low in large grain cultivar, but that of stem in Tamjinbyeo, SR7796F$_{7}$ and SR11269F$_{6}$ was 36%, 38% and 42%, respectively. In canopy architecture leaf blade weight was the highest 30~40cm, 40~50cm and 70~8Ocm above the ground in SR11269F$_{6}$, Tamjinbyeo and SR7796F$_{7}$, respectively. Range of panicle distribution in the canopy was 40cm in Tamjinbyeo, and was 70cm in large-grain cultivars. The results suggest that it has to take a growing interest in developing culture method for increasing LAI during growth stage, especially, around heading, in keeping green leaf till ripening stage and also in growing unformly plant within rice hill in large grain cultivars.cultivars.ltivars.

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Effect of $CO_2$ Concentration, NAEH and Light Intensity on the Photoautotrophic Growth of Campanula punctata 'Rubriflora' Plantlets In Vitro (자주초롱꽃의 기내 자가영양배양시 $CO_2$농도, 환기횟수 및 광도가 생장에 미치는 영향)

  • Shim, Jae-Nam;Kim, Gyeong-Hee;Jeong, Byoung-Ryong
    • Journal of Bio-Environment Control
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    • v.14 no.4
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    • pp.233-238
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    • 2005
  • Growth of Campanula punctata 'Rubriflora' plantlets, as affected by three levels of photosynthetic photon flux (PPF), 70, 110, and $220{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, two levels of $CO_2$ concentration, 500 and $1,500{\mu}mol{\cdot}m^{-1}$, and two levels of number of air exchanges per hour (NAEH), 0.1 and $2.8 h^{-l}$, was studied. Explants were obtained from photomixotrophically-micropropagated plantlets. Four explants were planted in each $3.7{\times}10^{-4}m^3$ polycarbonate box containing MS basal medium and no added sucrose. Explants were cultured under cool-white fluorescent lamps for $16h{\cdot}d^{-1},\;at\;25\pm1^{\circ}C$ temperature, and $70\~80\%$ relative humidity In treatments of $2.8h^{-1}$ NAEH, a 10mm round hole made on the vessel cap was sealed with a microporous filter. For higher $CO_2$ concentrations in the culture room, $CO_2$ gas was provided from a tank of liquefied $CO_2$. Fresh and dry weights, length of the longest root, and number of leaves significantly increased with increasing PPF and especially $CO_2$ concentration. Length of the longest root, number of leaves, fresh and dry weights, and chlorophyll concentration were enhanced with increased NAEH. However, leaf area was the smallest in the $220{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}\;PPF\;2.8h^{-1}$ NAEH and especially, $1,500{\mu}mol{\cdot}mol^{-1}\;CO_2$ concentration treatment. Treatment effect became more produced with time. Overall, treatment with $220{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}\;PPF\;and\;1,500{\mu}mol{\cdot}mol^{-1}\;CO_2$ gave the most vigorous growth.

Studies on the In Vitro Development of Cloned Embryos by Somatic Cell Nuclear Transfer in Korean Native Goats (재래산양의 체세포 핵이식에 의한 복제수정란의 체외발달에 관한 연구)

  • Park H. S.;Kim T. S.;Jung S. Y.;Lee Y. H.;Jung J. Y.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.105-112
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    • 2005
  • The present study was conducted to examine some factors affecting in vitro development of oocytes from somatic cell nuclear transfer (SCNT) in Korean native goats. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean Native goats. For nuclear transfer, the fibroblasts from caprine ear cells and fetal fibroblasts were surgically harvested and were cultured in vitro until cell confluency in serum-starvation condition (TCM-199 + $0.5\%$ FBS) for 3 to 5 days. The zona pellucidae of matured oocytes were partially drilled by laser irradiation. A single somatic cell was individually transferred into each enucleated oocyte. The reconstructed oocytes were then electrically fused and activated. Activated NT embryos were cultured in mSOF medium supplemented with $0.8\%\;BSA\;6\~7\;day\;at\;39^{\circ}C,\;5\%\;CO_2,\;5\%\;O_2,\;90\%\;N_2$ in air. There were no significant difference in the number of embryos cleaved and 4-cell development between the fibroblast nuclei from mature ear cells and fetal cells, but the rate of 8-cell development was higher (P<0.05) in ear cells $(40.5\%)$ than in fetal cells $(55.5\%)$. However, the embryo development to morula or blastocyst was not significantly different between both the groups$(6.7\%\;vs\;16.0\%)$, respectively. The number of embryo cleaved $(79.0\%)$ were higher (P<0.05) in the oocytes activated with ionomycin+6-DMAP than in the oocytes activated electrically $(9.5\%)$. The development of fused embryos to morula or blastocyst was found $15.6\%$ in ionomycin+6-DMAP, but no morula or blastocysts were developed in electrical stimulation. The development rate of SCNT embryos to morula or blastocyst was love. (P<0.05) in SCNT embryos $(19.0\%\;vs\;0.0\%)$ than that in parthenotes $(66.1\%\;vs\;59.1\%)$. In the parthenotes, the cleavage rate and development to morula or blastocyst were significantly higher (P<0.05) as $86.8\%\;and\;50.0\%$ in ovulated oocytes than in follicular oocytes $(69.0\%\;vs\;23.6\%)$, respectively. These results suggest that some factors Including superovulation treatment, oocyte source, maturation of follicular oocytes, activation method and culture condition may affect in vitro developmental capability of embryos produced by somatic cell nuclear transfer in Korean Native goats, and the fusion rate be greatly low compared with other species.

Sex Detection and In Vitro Development of Biopsied Bovine Embryo for LAMP Based Embryo Sexing (LAMP 방법에 의한 소 수정란의 성 판별과 Biopsy에 따른 수정란의 체외발달)

  • Cho S. R.;Choi S. H.;Kim H. J.;Han M. H.;Choe C. Y.;Chung Y. G.;Son D. S.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.169-176
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    • 2005
  • Loop-mediated isothermal amplification (LAMP) is novel DNA amplification methods that amplifies a target sequence specifically under isothemal condition. The present study was to assess the in vitro viability afier biopsy and sexing rate of different types of embryo biopsied. In vivo compact morulae and blastocyst embryos were obtained from Korean Native Cow (KNC) superovulated with FSH (Antorin, R-10) on 7 Day after artificial insemination. in vitro compact morulae and blastocyst embryos were obtained with KNC or Holsteins that were gained on 6, 7 or 8 day after in vitro fertilization(IVF) with frozen semen. Biopsy of bovine embryo was carried out in a $80{\mu}l$ drop with $Ca^{2+}-Mg^{2+}$ free D-PBS and the viability of biopsied embryos were evaluated in IVMD (IFP, Japan) medium at 12 hrs culture time. The sex ratio of biopsied Hanwoo embryos were male vs. female of $43.5\%\;vs.\;56.5\%$ in vivo and $33.9\%\;vs.\;49.2\%$ in vitro respectively, and male rate of biopsied Holstein embryos were significantly higher than female $(70.8\;vs.\;29.2\%)$. and indefinite rate of in vitro embryos was $16.9\%$ and in vivo was not. The degeneration rate of biopsied embryo, in vitro embryos were significantly higher than in vivo $(13.2\%\;vs,\;0.0\%,\;p<0.05)$. The survivability of in vivo embryo were between biopsied following punching method was significantly (P<0.05) higher than bisection method produced embryos $(100\%\;vs.\;83.3\%)$ and in vitro had no difference. However, the degeneration rate of biopsied embryo by bisection method was significantly higher than punching methods between in vivo and in vitro $(16.7\;vs.\;22.6\%,\;respectively,\;p<0.05)$. In conclusion, these results indicate that punching method was optimal and survivability after embryo biopsy was useful for reducing the damage caused by the embryo biopsy procedure for LAMP-based embryo sexing.

Effects of FGF on Embryonic Development In Vitro in Hanwoo COCs (한우 난구 복합체의 체외발생에 있어서 FGF(Fibroblast Growth Factor)가 미치는 영향)

  • Choi S.H.;Cho S.R.;Kim H.J.;Choe C.Y.;Han M.H.;Son D.S.;Chung Y.G.;H. Hoshi
    • Journal of Embryo Transfer
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    • v.21 no.2
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    • pp.157-162
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    • 2006
  • It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs (cumulus oocytes complexes) in vitro. Attempts had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of FGF (fibroblast growth factor) on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in HPM 199 (Inst. of Functional peptide, Japan) containing 0.1, 1 and 10 ng/ml FGF for 24 hr, maturation rates to metaphase II ($70.0{\sim}75.0%$) were significantly higher (p<0.05) than that of control group (0 ng/ml FGF, 37.5%). When matured COCs with FGF were cultured in maturation medium after in vitro fertilization, developmental rates to blastocysts were 9.5, 0 and 2.9%, respectively, compared to 25.0% of the control group (p<0.05). When the matured COCs with FGF were cultured in HPM 199 (IFP971, Inst. of Functional peptide, Japan) containing 10% FBS, 0.8% BSA or 0.1% PVA (polyvinyl alcohol), the blastocyst formation rates were 12.4, 12.8 and 8.5%, respectively, while the rates of matured COCs with FGF and cultured with IVMD and IVD (Inst. of Functional peptide, Japan) without serum were 38.4% and 34.8%, respectively (p<0.05). These results suggested that FGF is available for in vitro maturation of bovine COCs and is not suitable for in vitro development, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine embryo development.

Characterization of Bacteriocin Produced from Isolated Strain of Bacillus sp. (Bacillus 속 분리주가 생산하는 박테리오신의 특성 조사)

  • Ham, Seung-Hee;Choi, Nack-Shick;Moon, Ja-Young;Baek, Sun-Hwa;Lee, Song-Min;Kang, Dae-Ook
    • Journal of Life Science
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    • v.27 no.2
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    • pp.202-210
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    • 2017
  • As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

Kinetic Studies of Lactic Acid Fermentation (Part 3) Effect of Phenol Derivatives on Fermentation (유산균발효에 관한 동력학적 연구 (제3보) 발효에 미치는 Phenol 유도체의 영향)

  • LEE Keun-Tai;YANG Hyeun-Suk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.14 no.4
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    • pp.212-216
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    • 1981
  • The growth of Lactobacillus bulgaricus treated with vanillin, ortho-vanillin and guaiaco1 was studied on synthetic medium in mechanically agitated chemostat culture, The exponential-phase growth rate exhibited a maximum at the cells treated with 50 ppm vanillin. That stimulation, however, appears to be an effect on growth rate rather than total cell growth. And the others were inhibited by the chemicals. Much greater inhibition in growth of the cells treated with 100 ppm of each chemical than oars treated with 50 ppm was observed after 25 hour fomentation. For aerobic microbes, the alcohol dehydrogenase reaction is enhanced for the reproduction of NAD, which consequently cause to stimulate fermentation. For micro-aerophilic microbes , however, the same effect was not observed at the present study at least in the case of cell concentration. However except f or one treated with 50 ppm vanillin the same effect was observed in the case of growth is to. From the result using the glucose as a substrate, it was found that the cell concentrations measured in terms of ultimate optical density (UOB/ml), were 0.96 and 0.92, when treated with 50 and 100 ppm vanillin; 0.40 and 0.45 when treated with ortho-vanillin 50 and 100 ppm: 0.49 and 0.47, when treated with guaiacol 50 and 100 ppm. The specific growth rates were 0.44, 0.15, 0.25, 0.29, 0.37, and 0.34; the specific production rates wire 0.33, 0.15, 0.16, 0.22, 0.28, and 0.26 and the glucose concentrations (g/1) after 25 hour fermentation were 23.5, 32.8, 31.5, 29.5, 28.0 and 28.8, these all in the same sequences as the first.

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New horizon of geographical method (인문지리학 방법론의 새로운 지평)

  • ;Choi, Byung-Doo
    • Journal of the Korean Geographical Society
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    • v.38
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    • pp.15-36
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    • 1988
  • In this paper, I consider the development of methods in contemporary human geography in terms of a dialectical relation of action and structure, and try to draw a new horizon of method toward which geographical research and spatial theory would develop. The positivist geography which was dominent during 1960s has been faced both with serious internal reflections and strong external criticisms in the 1970s. The internal reflections that pointed out its ignorance of spatial behavior of decision-makers and its simplication of complex spatial relations have developed behavioural geography and systems-theoretical approach. Yet this kinds of alternatives have still standed on the positivist, geography, even though they have seemed to be more real and complicate than the previous one, The external criticisms that have argued against the positivist method as phenomenalism and instrumentalism suggest some alternatives: humanistic geography which emphasizes intention and action of human subject and meaning-understanding, and structuralist geography which stresses on social structure as a totality which would produce spatial phenomena, and a theoretical formulation. Human geography today can be characterized by a strain and conflict between these methods, and hence rezuires a synthetic integration between them. Philosophy and social theory in general are in the same in which theories of action and structural analysis have been complementary or conflict with each other. Human geography has fallen into a further problematic with the introduction of a method based on so-called political ecnomy. This method has been suggested not merely as analternative to the positivist geography, but also as a theoretical foundation for critical analysis of space. The political economy of space with has analyzed the capitalist space and tried to theorize its transformation may be seen either as following humanistic(or Hegelian) Marxism, such as represented in Lefebvre's work, or as following structuralist Marxism, such as developed in Castelles's or Harvey's work. The spatial theory following humanistic Marxism has argued for a dialectic relation between 'the spatial' and 'the social', and given more attention to practicing human agents than to explaining social structures. on the contray, that based on structuralist Marxism has argued for social structures producing spatial phenomena, and focused on theorising the totality of structures, Even though these two perspectives tend more recently to be convergent in a way that structuralist-Marxist. geographers relate the domain of economic and political structures with that of action in their studies of urban culture and experience under capitalism, the political ecnomy of space needs an integrated method with which one can overcome difficulties of orthhodox Marxism. Some novel works in philosophy and social theory have been developed since the end of 1970s which have oriented towards an integrated method relating a series of concepts of action and structure, and reconstructing historical materialism. They include Giddens's theory of structuration, foucault's geneological analysis of power-knowledge, and Habermas's theory of communicative action. Ther are, of course, some fundamental differences between these works. Giddens develops a theory which relates explicitly the domain of action and that of structure in terms of what he calls the 'duality of structure', and wants to bring time-space relations into the core of social theory. Foucault writes a history in which strategically intentional but nonsubjective power relations have emerged and operated by virtue of multiple forms of constrainst wihthin specific spaces, while refusing to elaborate any theory which would underlie a political rationalization. Habermas analyzes how the Western rationalization of ecnomic and political systems has colonized the lifeworld in which we communicate each other, and wants to formulate a new normative foundation for critical theory of society which highlights communicatie reason (without any consideration of spatial concepts). On the basis of the above consideration, this paper draws a new norizon of method in human geography and spatial theory, some essential ideas of which can be summarized as follows: (1) the concept of space especially in terms of its relation to sociery. Space is not an ontological entity whch is independent of society and has its own laws of constitution and transformation, but it can be produced and reproduced only by virtue of its relation to society. Yet space is not merlely a material product of society, but also a place and medium in and through which socety can be maintained or transformed.(2) the constitution of space in terms of the relation between action and structure. Spatial actors who are always knowledgeable under conditions of socio-spatial structure produce and reproduce their context of action, that is, structure; and spatial structures as results of human action enable as well as constrain it. Spatial actions can be distinguished between instrumental-strategicaction oriented to success and communicative action oriented to understanding, which (re)produce respectively two different spheres of spatial structure in different ways: the material structure of economic and political systems-space in an unknowledged and unitended way, and the symbolic structure of social and cultural life-space in an acknowledged and intended way. (3) the capitalist space in terms of its rationalization. The ideal development of space would balance the rationalizations of system space and life-space in a way that system space providers material conditions for the maintainance of the life-space, and the life-space for its further development. But the development of capitalist space in reality is paradoxical and hence crisis-ridden. The economic and poltical system-space, propelled with the steering media like money, and power, has outstriped the significance of communicative action, and colonized the life-space. That is, we no longer live in a space mediated communicative action, but one created for and by money and power. But no matter how seriously our everyday life-space has been monetalrized and bureaucratised, here lies nevertheless the practical potential which would rehabilitate the meaning of space, the meaning of our life on the Earth.

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