• KSBB Journal
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• v.23 no.3
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• pp.205-212
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• 2008

The efficient soluble expression of human epidermal growth factor (hEGF) was achieved by using functional fusion partners in cytoplasm and periplasm of Escherichia coli (E. coli). hEGF was over-expressed in inactive inclusion body form in cytoplasm of E. coli due to improper disulfide bond formation and hydrophobic interaction, yielding about 5.9 mg/L in flask culture. Six functional fusion partners were introduced by linking to N-terminal part of hEGF gene for the high-level expression of soluble and active hEGF in cytoplasm and peri plasm region. Three fusion partners for cytoplasmic expression such as acidic tail of synuclein (ATS), thioredoxin (Trx) and lipase, and three fusion partners for periplasmic expression such as periplasmic cystein oxidoreductases (DsbA and DsbC) and maltose binding protein (MBP) were investigated. hEGF fused with ATS and DsbA showed over 90% of solubility in cytoplasm and periplasm, respectively. Especially DsbA was found to be an efficient fusion partner for soluble and high-level expression of hEGF, yielding about 18.1 mg/L and three-fold higher level compared to that of insoluble non-fusion hEGF in cytoplasm. Thus, heterologous proteins containing complex disulfide bond and many hydrophobic amino acids can effectively be produced as an active form in E. coli by introducing a suitable peptide or protein.

• Korean Journal of Plant Tissue Culture
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• v.24 no.1
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• pp.1-35
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• 1997

Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.63-71
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• 2004

The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmental facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Juniperus chinensis xylem extract on the UV and SLS-induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. We found that Juniperus chinensis xylem extracts had potent radical scavenging effect by 98％ at 100 $\mu\textrm{g}$/mL. Fluorometric assays of the proteolytic activities of matrix metalloproteinase-l(MMP-1, collagenase) were performed using fluorescent collagen substrates. UV A induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97％ at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79％ at 25 $\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. In this test Juniperus chinensis decreased expression of interleukin 6 about 30％. Expression of prostaglandin E$_2$, (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay (EIA) using PGE$_2$ monoclonal antibody. At the concentrations of 5-50 $\mu\textrm{g}$/mL of the extracts, the production of PGE$_2$ by HaCaT keratinocytes (24 hours after 10 mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p〈0.05). The viability of cultured HaCaT keratinocytes was significantly reduced at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB irradiation, but the presence of these extracts improved cell viability comparing to control after UVB irradiation. We also investigated the protective effect of this extract in sodium lauryl sulfate (SLS)-induced irritant skin reactions from 24 hour exposure. Twice a day application of the extract for reducing local inflammation in human skin was done. Irritant reactions were assessed by various aspects of skin condition, that is, erythema (skin color reflectance) and transepidermal water loss (TEWL). After 5 days the extract was found to reduce SLS-induced skin erythema and improve barrier regeneration when compared to untreated symmetrical test site. In conclusion, our results suggest that Juniperus chinensis can be effectively used for the prevention of UV and SLS-induced adverse skin reactions such as radical production, inflammation and skin cell damage.

• Tuberculosis and Respiratory Diseases
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• v.57 no.6
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• pp.528-534
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• 2004

Background : Microscopic examination of sputum smears for acid-fast bacilli (AFB) is the most important and rapid diagnostic test for pulmonary tuberculosis. However, the AFB observed on the smear may represent either M. tuberculosis or nontuberculous mycobacteria (NTM). This study examined the clinical usefulness of a polymerase chain reaction test for M. tuberculosis (TB-PCR) for the differentiation of pulmonary tuberculosis and NTM lung disease in patients with smear-positive sputums in a tertiary hospital in Korea. Material and Methods : From January, 2003 to December, 2003, 826 AFB smear-positive and culture-positive sputum specimens were collected from 299 patients. Results : NTM were recovered from 26.6% (220/826) of the smear-positive sputum specimens and 23.4% (70/299) of the patients with smear-positive sputum. All the patients with isolated NTM had clinically significant NTM lung disease; 38 patients (54.3%) had M. avium and 26 patients (37.1%). had M. abscessus. In the patients with pulmonary tuberculosis, 78.7% of the patients (74/94) showed TB-PCR positivity, and all the patients with NTM lung disease showed negative results on the TB-PCR test (p<0.001). A positive result of the TB-PCR test on the sputum or bronchial washing fluid specimens was able to predict pulmonary tuberculosis with 88.4% sensitivity, 100% specificity, a 100% positive predictive value and a 79.7% negative predictive value for the patients with smear-positive sputum. Conclusion : The TB-PCR test for sputum specimens or bronchial washing fluid specimens could be useful for differentiating pulmonary tuberculosis and NTM lung disease for the patients with smear-positive sputum in Korea.

• Journal of Korean Society of Forest Science
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• v.101 no.3
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• pp.387-394
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• 2012

The study was conducted to develop an effective forest resources use models for an alternate use of abandoned quarry by an attitude survey. According to the result of survey, a pessimistic view due to dust, noise pollution, and forest damage was 5% higher than an affirmative view by economic benefits from the development of quarry. The 42% of the respondents preferred the alternate use of abandoned quarry and the 25% of the respondents wanted an art and cultural space. The optimum size of alternate use was 5-10 ha (43%) with the requirement of nearby residents (32%). According to the SWOT analysis for abandoned quarry, the strength factors were an effective use of land, the content development of modern industrial inheritance + cultural and art fusion, attraction for nearby city and visitors, a harmony of beauty landscape and clean environment, and a sustainable increase of domestic and foreign visitors with the 5-day-work week. The opportunity factors were the improvement of traffic networks through KTX and local highway, the creation of the new growth engines with the establishment of artistic creation belts, the providing of unique cultural and art space through grafting of tour and education, the creation of local income through stone processed goods, and the vitalization of local development through eco-city. The weakness factors were a psychological remoteness and backwardness, and the weakness of staying tour infra. The threat factors were a poor financial support for sustainable development in nearby quarry and a modify of legal and institutional system for the alternated use of abandoned quarry. The developed restoration models for the alternate use in abandoned quarry are classified to a sculpture park, a waterfall and lake park, a rock-climbing, a sports park + forest park, a native botanical garden, a culture and art park, a complex park, a water storage site, a water storage site to extinguish forest fire, a geriatric hospital, an agricultural facility, and a school site types etc. The results suggest that the alternate use in the abandoned soil and stone quarry is needed to establish facility use models with consideration of user's preference.

• Cartoon and Animation Studies
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• s.49
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• pp.359-382
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• 2017

Content is the complex of art and technology of trend, so it is important to experience different technologies for content education. Today, many non-profit organizations plan and operate numbers of programs for disabilities, low-income, and minority families to enhance the quality of life and the realization of social integration. These programs are limited to museums and galleries, not so pro-actively in progressing. Various contend education is necessary to the expansion of cultural exchange for the culturally alienated area. Naver is running an outreach program named . It is an experience-based outreach program where current cartoon / webtoon writers come directly to the school to inform students about the basic story of comics and comic techniques. However, the fact that the is not centered on the marginalized area but is centered on the Seoul Gyeonggi area, has the limitation that they can not benefit from a wide range of programs because they have a space limit of 'school', and, has a spatial limitation that the experience of the work is excluded. 'Outreach programs in marginalized areas' must be reorganized into a fluid dimension, not a fixed, single-system program. You should be able to experience and experience your work by directly using various professional equipment of comics based on your capacity and experience, local culture, religion, and society. These program participants will gain the effect of attractive and effective learning with empathy with their comic experience. Meanings of Comics content outreach program are following: First, the rich cultural archive can be used efficiently by providing various contents to existing outreach programs with the educational limitation of museums and galleries. Second, Comics contents can be enjoyed as a part of our life by understanding diversity and technology of contents. Third, because it is the program of expertise' participation, it can remodel, and restructure the severed experience in remote areas for the continuous growth and development, and furthermore, it can enhance the understanding of society.

• Korean Journal of Weed Science
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• v.31 no.3
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• pp.279-288
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• 2011

To suggest the weed management technique for environment friendly rice cultivation, we investigated occurrence patterns of weeds, the actual condition of weed management, and rice yield at the environment friendly agricultural complex located in Honam and Chungnam regions. The practical performance of weed management was relatively satisfactory in decreasing order of agricultural technique with golden-apple-snail (GAS) > agricultural technique with duck (Duck) > agricultural technique with rice bran (RB) > agricultural technique with soft-shelled turtle (ST). In the rice fields employed by agricultural technique with GAS, the dominant weeds were Echinochloa crus-galli, Ludwigia prostrata, Monochoria vaginalis, Sagittaria trifolia, and Aneilema keisak. However, E. crus-galli, M. vaginalis, L. prostrata, Aeschynomene indica and Bidens frondosa were found as dominant weeds at the fields using the Duck and E. crus-galli, M. vaginalis, L. prostrata, Polyganum hydropiper and Eleocharis kuroguwai at the fields using RB. In comparison of rice yield ($5.2\;MT\;ha^{-1}$) obtained from the conventional cultivation using herbicides, about 93% was reached by Duck, about 91% by GAS, about 92% by RB, and about 78% by ST. When rice qualities obtained from environment friendly rice cultivation were compared with those from the conventional cultivation, the producing rates of perfect kernel, immature kernel, immature opaque kernel, cracked rice, and damaged kernel were lower in the former cultivation, whereas contents of protein, amylose, and fatty acid were similar in the two cultivation methods. The problems found in the environment friendly agriculture were poor plowing and harrowing, carless irrigation management, and geological poor condition as cultivation area with cold water. These have caused severe infestation of weeds, frequent incident of disease and insect pest, and rice lodging. This resulted in reduction of rice yield as high as about 32 to 79% as compared with the conventional cultivation using herbicides.

• Development and Reproduction
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• v.6 no.1
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• pp.55-66
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• 2002

Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

• 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.