대장 염증 발생 과정에서 Ninjurin1 단백질의 대장 보호 기능에 관한 연구

학위논문 (박사) -- 서울대학교 대학원 : 약학대학 약학과, 2020. 8. 서영준.Disruption of colonic homeostasis caused by aberrant M1/M2 macrophage polarization and dysbiosis contributes to inflammatory bowel disease (IBD) pathogenesis. However, the molecular factors mediating colonic homeostasis are not well characterized. Here, I found that Ninjurin1 (Ninj1) limits colon inflammation by regulating macrophage polarization and microbiota composition under homeostatic conditions and during colitis development. Ninj1 deletion in mice induced hypersusceptibility to colitis, with increased prevalence of colitogenic Prevotellaceae strains and decreased immunoregulatory Lachnospiraceae strains. Upon co-housing with WT mice, Ninj1-/- mice showed increased Lachnospiraceae and decreased Prevotellaceae abundance, with subsequent improvement of colitis. Under homeostatic conditions, M1 macrophage frequency was higher in the Ninj1-/- mouse colons than WT mouse colons, which may contribute to increased basal colonic inflammation and microbial imbalance. Following colitis induction, Ninj1 expression was increased in macrophages; meanwhile Ninj1-/- mice showed severe colitis development and impaired recovery, associated with decreased M2 macrophages and escalated microbial imbalance. In vitro, Ninj1 knockdown in mouse and human macrophages activated M1 polarization and restricted M2 polarization. Finally, the transfer of WT macrophages ameliorated severe colitis in Ninj1-/- mice. These findings suggest that Ninj1 mediates colonic homeostasis by modulating M1/M2 macrophage balance and preventing extensive dysbiosis, with implications for IBD prevention and therapy.INTRODUCTION 1 1. Macrophage plasticity and polarization 1 2. Macrophages and intestine homeostasis 4 3. Macrophage phenotype in IBD 7 4. Gut microbiota and immune homeostasis 10 5. Dysbiosis and IBD 13 6. Ninjurin1 16 7. Dextran sulfate sodium (DSS)-induced colitis 19 PURPOSE OF THIS STUDY 22 MATERIALS AND METHODS 23 1. Mice 23 2. DSS-induced colitis 23 3. Histopathological Score 24 4. Antibiotic treatment 25 5. Bacterial 16S rRNA sequencing and data analysis 25 6. Co-housing experiment 27 7. Tissue harvesting 27 8. Preparation of bone marrow-derived macrophages (BMDMs) 27 9. Isolation of colonic lamina propria (cLP) cells 28 10. THP-1 cell culture and macrophage differentiation 29 11. Small interfering RNA (siRNA) Transfection 29 12. Macrophage stimulation 30 13. Immunofluorescence 30 14. Flow cytometry 31 15. RNA isolation and qPCR 32 16. Immunoblotting 32 17. Enzyme-linked immunosorbent assay (ELISA) 33 18. Adoptive transfer of macrophages 33 19. Data analysis and statistics 34 RESULTS 38 1. Enhanced susceptibility of Ninj1-/- mice to colitis is dependent on gut microbiota 38 2. Alteration of the gut microbiome by Ninj1 deficiency 45 3. Reconstitution of Ninj1-/- mice by WT fecal microbiota attenuates colon inflammation 50 4. Host Ninj1 deficiency and gut dysbiosis shaped by Ninj1 deficiency cooperatively accelerate severe colitis development 58 5. Ninj1 deficiency establishes pro-inflammatory colonic environment by increasing the numbers of M1 macrophages 63 6. Ninj1 deficiency increases inflammatory response to bacteria in macrophages via activation of chronic inflammatory signaling in vitro 75 7. Ninj1-/- mice exhibited impaired colitis recovery 81 8. Reduction of M2 macrophages in Ninj1-/- mice delays colitis recovery 86 9. Ninj1 restricts M1 macrophage polarization but activates M2 polarization in vitro 92 10. Transfer of WT macrophages ameliorated severe colitis development of Ninj1-/- mice 99 DISCUSSION 106 REFERENCES 114 요약 (국문초록) 129Docto

Critical Thinking and Female Values

If critical thinking is unfair to women, it should not be an educational ideal. First, this paper considers the claims by Gilligan, Tannen, Nye, and McClintock that critical thinking seeks after male values such as logic, form and abstraction. I argue that it is possible to impart critical thinking with female values such as love and cooperation in addition and that such critical thinking must be educated. Critical Thinking is needed when we interpret charitably an argument as well as we find fault with it. Female values work importantly in such processes

The causal efficacy of mental properties : an epiphenomenal interpretation of nonreductive physicalism

학위논문(박사)--서울대학교 대학원 :철학과서양철학전공,2000.Docto

암모니아에 의한 질소산화물의 선택적 촉매 환원에서 구리이온이 교환된 제올라이트 촉매가 담지된 하니콤반응기 설계

Docto

콜드 프레스 공정을 이용한 음극지지체 GDC/YSZ 전해질 이중층 고체산화물 연료전지 연구

학위논문 (박사)-- 서울대학교 대학원 : 기계항공공학부, 2014. 2. 차석원.The fact that hydrogen will be the last energy source became no more attractive to us. The main issue is which kind of energy conversion device will be going to survive in the future. Since many researchers highlighted the fuel cell as the next generation power source, a lot of researched have been conducted to commercialize it. SOFCs have many advantages in comparison with typical PEMFCs which have shown water management problem, usage of novel catalyst, patent issue for polymer electrolyte, expansive graphite bipolar plate and CO poisoning. So many researchers in energy field have been thought SOFC would be the promising device. But the main bottle neck for the commercialization of SOFC has been its high operation temperature. It can cause thermal mismatch between MEA, nickel agglomeration, reactions between component materials, restricted sealant choice and expensive interconnecter material. So we focused our interest to IT-SOFC. Its temperature position can avoid many problems of HTSOFC and LT-SOFC maintaining the competitiveness of original SOFCs characteristics. Moreover, process cost issue about compaction, sintering and more complicated high temperature process is one of the bottle-neck for commercialization. In this study, we studied the lower the operating temperature in order to solve these problems, while reducing the thickness of the electrolyte and dropping the number of steps compared to the conventional method. First, we prepared the bi-layer electrolyte solid oxide fuel cell which was deposited yttria-stabilized zirconia (YSZ) and gadolinia dopted ceria (GDC) which is high ionic conductivity material at low and intermediate temperature. This structure is verified that high performance, sufficient durability and operation in a low temperature. It is confirmed the successful deposition of YSZ through the scanning electron microscope (SEM) and X-ray photoelectron spectroscopy (XPS). We obtained a 50% improvement power density and 5% higher open circuit voltage (OCV) than the output of pure GDC electrolyte cell at 600 degrees Celsius. We confirmed that the YSZ layer prevent enhancing electronic conductivity and micro crack which can make voltage-drop and enlarge ohmic loss in order to reduce GDC layer by SEM image and EIS. Second, because we verified effect of YSZ/GDC bi-layer electrolyte structure to the performance, it is studied about bi-layer electrolyte anode supported type solid oxide fuel cell structure which could have higher power density. Various wet ceramic processes and thin film deposition processes are employed to electrolyte deposition method to calcined anode substrate. Finally, we established cold press process that can sinter from substrate to electrolyte at one step process. In order to enhance the controllability and uniformity of thickness of the layer, spray dry coating method that can control about micron range using powder vehicle is employed. The compressed dry powder substrate-bi-layer electrolyte was co-sintered, and we fabricated LSCF-GDC cathode using screen printing method. It is possible as compared with the wet method, to reduce the sintering step one or more times, forming method such a great advantage in time and cost. We obtained power density of 210mW/cm2 at 600 degrees Celsius, and 409mW/cm2 at 800 degrees Celsius from prepared bi-layer electrolyte cell. It is 0.1V higher open circuit voltage and 15% higher maximum power density rather than one of non bi-layer electrolyte cell. Third, we have improved the fabrication process to increase the performance of bi-layer electrolyte cell via cold press process. In order to make porous anode substrate, the PMMA powder was mixed with anode substrate powder which is used previous bi-layer electrolyte cell fabrication. And it is achieved the power density is 460mW/cm2 at 700 degrees Celsius. Further, by adding one functional layer between the anode substrate and electrolyte layer, to complement the reduction in mechanical strength that can occur in the case of porous anode support, to improve the electrochemical performance by cold press process. As a result, it was possible to obtain a power density and mechanical strength is enhanced, and is more stably maintained even during operation. Finally, we modeled and calculated the compaction behavior during cold press process using COMSOL multiphysics. The Surface von Mise stress was predicted using the Drucker-Prager Cap model. Through this model, we were able to determine at the time of production to minimize the damage, and the condition of the powder. In addition, through the modeling, the basic direction for the production of large-scale cells could be present.Chapter 1. Introduction 1 1.1 Research Background………………………………………….…………….1 1.2 Solid oxide fuel cell ………………………...………………….……………5 1.3 The operating temperature of SOFCs…………………………….………….8 1.3.1 Reducing the thickness of electrolyte……………………………………11 1.3.2 High conductivity electrolytes material…………………………………12 1.4 Bi-layered electrolytes SOFCs………………………………….…………..14 1.5 The supported type of SOFC………………………………….…………….19 Chapter 2. Cell fabrication and experimental process 22 2.1 Cell fabrication and test for verification of YSZ-GDC bi-layer……….……22 2.1.1 The components of bi-layered electrolyte………………………….…...24 2.1.2 ALD-YSZ for a functional layer………………………………….…….25 2.1.3 Cell fabrication…………………………………………………….……27 2.1.4 Cell test procedure……………………………………………….……..30 2.1.5 Result of ALD-YSZ/GDC bil-layered cell test………………….……...38 2.2 Approach of various anode-supported type SOFC cell fabrication process ...………………………………………………………………………….48 2.2.1 Literature survey………………………………………………….…….49 2.2.3 Cold press process………………………………………………….…..52 2.2.4 Summary of cell fabrication process and Cell test procedure …………54 Chapter 3. Test of the YSZ/GDC bi-layered electrolyte anode supported type cell via cold press process 63 3.1 Introduction………………………………………….………………………63 3.1.1 Literature survey………………………………….…………………….64 3.2 Research Methodology…………………………..………………………….65 3.3 Results…………………………………………..………….………………..69 3.4 Conclusion………………………………………………….……………….75 Chapter 4. Anode functional layer for porous anode substrate 76 4.1 Introduction…………………………………………………………………76 4.2 Porous anode substrate cell……………………………………..………..….77 4.2.1 Porous anode substrate cell fabrication…………………….….……..…77 4.2.1 Cell fabrication……………………………………………….…………78 4.2.2 Cell test result………………………………………….……………….79 4.3 Anode functional layer cell……………………………………….…………82 4.3.1 Fabrication…………………………………………………….………..82 4.3.2 Experimental…………………………………………………..………..85 4.3.3 Result………………………………………………………..……….…86 4.3.4 Discussion……………………………………………………..………..91 Chapter 5. Numerical analysis of behavior of fabrication process by finite element method 93 5.1 Introduction……………………………………………………….…………93 5.2 Geometric description…………………………………………….…………94 5.3 Model description……………………………………………….…………..96 5.4 Analysis result………………………………………………………………98 Chapter 6. Concluding remarks 102 6.1 Conclusion………………………………………………………………...102 6.2 Future works………………………………………………………………105 List of figures Fig 1.1 Operation principle of a fuel cell [1] 4 Fig 1.2 Schematics of PEMFC, MCFC and SOFC [5] 7 Fig 1.3 Advantages of SOFCs 7 Fig 1.4 Competitiveness of IT-SOFCs 10 Fig 1.5 Comparison of conductivity of various materials 13 Fig 1.6 The various approaches about bi-layer cell fabrication process[23,47,75] 18 Fig 1.7 Types of mechanical supported cell. (a) the cathode supported type (b) the electrolyte supported type (c) the anode supported type 21 Fig 2.1 Schematic of a YSZ/GDC bi-layered electrolyte SOFC 23 Fig 2.2 Example of ALD-YSZ based μSOFC [56] 26 Fig 2.3 Schematics of the ALD-YSZ / GDC cell and the pure GDC cell and actual image of the anode and cathode sides 29 Fig 2.4 ALD-YSZ bi-layered cell fabrication procedure 29 Fig 2.5 Button cell test station and P&ID diagram 32 Fig 2.6 Cell pressing unit and indicator 33 Fig 2.7 MFCs and controllers 34 Fig 2.8 Solartron SI 1287 and SI 1260 34 Fig 2.9 Experiment preparation process on the fifth design fixture. 37 Fig 2.10 XPS spectra on the surfaces of ALD-YSZ / GDC samples (red) before the electrode sintering process and (blue) after the cell operation 40 Fig 2.11 XPS depth profile of the bi-layered electrolyte which was 345 cycles (50nm) ALD-YSZ coated GDC 40 Fig 2.12 OCV transitions of the pure GDC cell and of the 690 cycles (100nm) ALD-YSZ / GDC cell at 620°C using 5% H2 fuel during the anode reduction step 42 Fig 2.13 iV and power density curves at 600°C for the pure GDC cell, 50nm ALD-YSZ / GDC cell, and the 100nm ALD-YSZ / GDC cell 45 Fig 2.14 Comparison of the iV and power density at 600°C as the hydrogen and air feed rates increased from 200sccm to 300sccm 45 Fig 2.15 EIS Nyquist spectra at 600°C (a) for the 690 cycles (100nm) ALD-YSZ / GDC cell and (b) for the pure GDC cell 47 Fig 2.16 ALD bi-layered SOFC SEM cross sectional, power and IV performance[39] 51 Fig 2.17 Wet cermic process bi-layered SOFC SEM cross sectional, power and IV performance[47] 51 Fig 2.18 Cold press single layer anode supported SOFC SEM cross sectional image[54] 53 Fig 2.19 Power density and IV curve of Electrolyte supported cell and Anode supported cell. 56 Fig 2.20 IV and power density of spray coated electrolyte cell, 600℃ humidified H2 : 100 SCCM N2 : 100 Air : 100 SCCM 60 Fig 2.21 Cross sectional SEM image of the spray coated electrolyte 60 Fig 2.22 Modified test fixture using gold ring sealant 62 Fig 3.1. (a) Scanning electron microscopy of cross-section of a YSZ/GDC bilayer electrolyte film on a anode substrate. (b) morphology of GDC electrolyte plane 66 Fig 3.2. Measured OCV vs temperature for single cells with 50 µm GDC monolayer electrolyte, 50 µm YSZ monolayer electrolyte, and 8µm YSZ/ 40µm GDC bilayer electrolyte, respectively. 67 Fig 3.3 (a) Cell voltage and (b) power density as function of current density of a YSZ (8 µm)/GDC (40 µm) bilayer electrolyte cell measured between 600 and 800 °C in humidified hydrogen and open air. 71 Fig 3.4. Cell voltage and power density vs. current density of a YSZ (8 µm) / GDC (40 µm) bilayer electrolyte cell measured 600 ◦C in humidified hydrogen and open air. 72 Fig 3.5 Result of electrochemical impedance spectroscopy of GDC single electrolyte cell and YSZ/GDC bilayer electrolyte cell, 600°C 74 Fig 4.1 IV and power density of PMMA porous substrate bi-layer electrolyte cell 80 Fig 4.2 EIS reslut of PMMA porous substrate bi-layer electrolyte cell 81 Fig 4.3 IV performance curve in various thickness of anode functional layer 89 Fig 4.4 Power density in various thickness of anode functional layer 89 Fig 4.5 Similarity of IV curve of dense anode substrate cell and thick anode functional layer cell 90 Fig 4.6 Peak power density of various thickness of anode functional layer 90 Fig 5.1 Schematic diagram of the substrate model 95 Fig 5.2. Drucker-Prager Cap model: yield surface in the p-q plane. 97 Fig 5.3Criterion stress of compaction processed cell 101 Fig 6. 1 Summary of whole test results in this study 104 List of tables Fig 1.1 Various researches about bi-layer cell process 17 Fig 2.1 Summary of fabrication approaches 59 Fig 2.2 The electrolyte slurry recipe for wet ceramic process 59 Fig 4.1 Various approach for fabrication of anode functional layer 84 Fig 4.2 Thickness of Anode functional layer 84 Fig 5.1 Dimension and global parameter of the substrate model 95 Fig 5.2 Properties of materials for FEM modeling 100Docto

제초제 저항성 벼 익산483과 밀양204의 대사체 연구

This study aimed to investigate the substantial equivalence of two genetically modified rice varieties, Iksan483 and Milyang204, developed as glufosinate-tolerant rice in Korea and the corresponding parental lines. In order to assess the compositional changes and identify potential unintended effects of the genetic modification, different strategies such as the non-targeted and targeted (compound-specific, nutritional composition) metabolite profiling approaches were applied. The volatile metabolic constituents in leaf and root were also investigated for possible environmental effect. The metabolite profiling in non-targeted approach was carried out using two kinds of sample preparation method (Method I and II). Moreover, the mass spectral library of expected metabolites in rice was produced including both mass spectra and retention index. The retention indices of total 191 metabolites (59 lipid and 132 polar metabolites) were calculated and used for identification of unknown components in this study. It was possible for simpler and faster preparation by method II than by method I. By method I, 64, 34, and 42 metabolites were identified in rice grain, leaf and root, respectively, whereas, 96, 98 and 51 metabolites were identified by method II. In principle component analysis, examination of the scores and loadings plots showed effects on the metabolite profile resulting from genetic modification of Iksan483 and Milyang204 were of lower magnitude than the effects of the genetic diversity or difference between conventional varieties. Comparative assessment of nutritional composition as the targeted approach was conducted with genetically modified rice grains and its conventional counterparts. Nutrients including proximates, fatty acids, amino acids, minerals, vitamins, and antinutrients were investigated using several statistical comparisons. The results showed that, except for small differences in a few fatty acids, minerals, and trypsin inhibitor, there was no significant difference between genetically modified rice and conventional counterpart variety with respect to most of their nutrient composition. Most of levels of nutrients were in good compliance with the literature ranges, showing substantial equivalency. The results of principle component analysis demonstrated that any differences between the genetically modified and conventional rice varieties are in range of the differences observed among conventional varieties grown in different years. The comparison between results of nutritional analysis and metabolite profiling suggested that metabolite profiling could be a valuable tool for qualitative assessment and appropriate for screening of the difference of metabolites in a brief time. The quantitative analysis of target components and cross-check must be conducted if the difference between treatments was observed in metabolite profiling. The volatile constituents in leaf and root of genetically modified rice and conventional rice were isolated by solvent-assisted flavor evaporation (SAFE). Various compounds (15~17) were identified in rice leaves, while 10~12 volatiles were identified in rice roots. The major volatiles were phenylacetaldehyde, 3-methylbutanoic acid, 2,3-butanediol, 2-hexenal, benzyl alcohol and hexanal. The PCA results indicated no evidence of any significant differences between the conventional and transgenic rice. Therefore, it was concluded that the herbicide tolerant rice varieties, Iksan483 and Milyang204, are substantially equivalent to their parental lines including Anjung, Shindongjin, Dongjin and Junam in aspects of metabolite, nutritional and volatile composition.본 연구에서는 국내에서 개발된 제초제 저항성 벼인 익산483과 밀양204이 이들 모품종과 실질적으로 동등하다는 것을 규명하고자 하였다. 이를 위해 유전자재조합 벼와 모품종인 안중벼, 신동진벼, 동진벼 그리고 주남벼의 현미, 잎, 뿌리를 대상으로 metabolite profiling을 이용하여 식물체내의 다양한 대사체를 비교해석하였으며, 중요한 영양성분 및 반영양성분 등을 정량분석하였다. 또한, 환경영향평가의 일환으로 잎과 뿌리의 휘발성 대사물질을 비교분석하였다. 제초제 저항성 벼의 metabolite profiling는 두 가지 전처리법(Method I, II)을 활용해 수행하였다. 보다 편리하고 정확한 대사체 동정을 위해 벼에서 존재할 것이라 예상되는 191개 대사체에 대한 mass spectra와 retention index 정보가 담긴 library를 자체 제작하였으며 본 연구에서 대사체를 동정하는데 사용하였다. 두 가지 전처리방법 중 Method II에 따른 전처리가 휠씬 빠르고 간단하였다. Method I를 이용한 metabolite profiling 결과, 현미, 잎, 뿌리에서 각각 64개, 34개, 42개의 대사체를 확인하였으며 Method II를 이용한 경우는 각각 96개, 98개, 51개 대사체가 동정되었다. 다변량 분석결과, 익산483과 밀양204의 대사체 조성은 일부 대사체를 제외하고 4개 모품종과 큰 유의적 차이를 보이지 않았다. 영양성분 비교분석은 일반조성(수분, 조단백, 조지방, 조섬유, 회분), 지방산, 미네랄, 비타민 그리고 phytic acid, lectine, trypsin inhibitor와 같은 반영양성분을 대상으로 수행하였다. 유전자재조합 벼와 모품종간의 동등성은 일부 지방산, 미네랄, trypsin inhibitor 성분을 제외한 대부분의 영양성분에서 증명되었다. 또한, 조사된 영양성분 함량은 기존 문헌에서 보고된 결과내에 존재하였다. 다변량분석결과, 계통보다는 재배환경에 의해 영양성분 조성이 더 큰 영향을 받았음을 확인하였다. 따라서, 익산483과 밀양204의 영양성분 조성은 유전자재조합에 의해 영향을 받지 않았다. Metabolite profiling 결과와 영양성분 분석결과 간의 비교를 통해, metabolite profiling은 정량분석보다는 정성분석에 더 적합하며 단시간내에 두 개체간의 대사체 차이를 확인하는데 더 큰 효용성이 있음을 확인하였다. Metabolite profiling에서 개체간의 대사체 차이를 확인하였다면, 해당 대사체의 정량분석을 통해 교차검정이 반드시 이루어져야 한다. SAFE 전처리방법을 통해 유전자재조합 벼와 모품종의 잎과 뿌리에서 휘발성 물질을 분석, 비교하였다. 잎에서는 15~17개의 휘발성분이 확인되었으며 뿌리에서는 10~12개 휘발성분이 검출되었다. 주요 휘발성분은 phenylacetaldehyde, 3-methylbutanoic acid, 2,3-butanediol, 2-hexenal, benzyl alcohol, hexanal 이었다. 다변량 분석결과, 제초제 저항성 벼와 모품종 사이에서 휘발성분 조성의 유의적 차이를 확인할 수 없었다. 결론적으로, 제초제 저항성 벼인 익산483과 밀양204는 모품종과 대사체, 영양성분, 휘발물질 조성측면에서 실질적으로 동등하였다.Docto

비소세포성 폐암세포에서 고리구조가 절단된 deguelin 유도체의 세포자살과 항 혈관신생 유도활성에 관한 연구

학위논문 (석사)-- 서울대학교 융합과학기술대학원 : 분자의학 및 바이오제약학과, 2015. 2. 김규원.Deguelin은 Mundeulea sericea를 포함하는 식물에서 추출한 rotenoid 화합물이며 구조적으로 5개의 고리(A, B, C, D, E)를 가지고 있다. 이들은 열 충격 단백질 90 (Hsp90)에 결합하여 이들의 기능의 억제를 통해 여러 암세포에서 증식억제를 유도한다고 알려져 있다. 열 충격 단백질 90은 암세포에서 과발현되어있으며 여러 가지의 종양단백질의 안정성에 기여한다고 알려져 있다. 그러나, deguelin은 이러한 항암효과에도 불구하고 정상세포에서 높은 독성으로 인해 임상적으로 사용되기 어려운 문제점을 가지고 있다. 그리하여 공동연구를 통해 합성된 deguelin 유도체인 SH-1242 화합물을 이용하여 이러한 부작용을 극복하고자 하였다. SH-1242는 deguelin 구조에서 B와 C 고리를 제거한 화합물이다. 비소세포성 폐암세포에서 SH-1242가 암세포 증식억제 활성 및 항 혈관신생 억제 그리고 그 기전에 대해서 연구하였다. 비소세포성 폐암세포와 정상 기관지 상피세포에서 SH-1242를 처리하였을 때, 세포 독성을 확인하기 위하여 MTT 분석을 수행하였다. 그 결과, 비소세포성 폐암세포에서는 농도 의존적으로 세포 독성을 나타내었으며, 정상 폐 상피세포에서는 deguelin과 비교하여 독성이 거의 없는 것으로 나타났다. 또한 SH-1242가 비소세포성 폐암세포 콜로니 형성에 어떠한 영향을 주는지 보기 위해 부착 의존적인 또는 비 의존적인 콜로니 형성 분석 실험을 진행하였다. 두 실험에서 SH-1242의 농도 의존적으로 콜로니 형성이 저해되었다. 이러한 암세포 증식억제는 세포자살과 관련되어지는지를 확인하기 위하여 유세포 분류기 분석과 western blot을 수행하였다. sub-G1기의 사멸세포가 증가되었고 caspase-3 신호전달경로가 억제되었다. 또한 SH-1242가 항 혈관신생억제 활성에 관여하는지 보기 위해서 western blot과 관 형성 분석을 수행하였다. 저산소상태에서 H1299 세포에 SH-1242를 처리하였을 때 저산소 유도 인자-1ɑ의 발현이 감소되었다. 관 형성 분석에서는 SH-1242를 처리하였을 때 matrigel위에 HUVEC 세포의 모세관 형성이 현저하게 줄어들었다. 이러한 암세포 사멸과 항 혈관신생억제가 열 충격 단백질 90의 기능 저하에 관여되는지를 조사하였다. 그 결과, SH-1242 농도 의존적으로 ErbB2, Akt, MEK, ERK 등 열 충격 단백질 90 클라이언트 단백질들의 발현이 억제되었다. 마지막으로 SH-1242가 H1299 또는 환자 유래 이식암 동물에서 세포 자살 또는 항 혈관신생 활성이 있는지 조사하였다. 그 결과, SH-1242를 처리한 암 조직에서 cleaved caspase3의 발현은 증가되었고 CD31의 발현은 감소되었다. 종합적으로, SH-1242가 정상 기관지 상피세포에는 독성이 없지만, 비소세포성 폐암세포에서 Hsp90의 억제를 통해 암세포의 증식과 혈관신생 활성을 억제하는 것으로 보여진다.ABSTRACT....................................... 1 LIST OF FIGURE..................................8 INTRODUCTION 1. Heat Shock protein 90 ..................... 9 1-1. Hsp90 in normal cellular homeostasis.....9 1-2. Hsp90 structure and function .......... 10 1-3. Hsp90 in cancer........................ 12 2. Hypoxia-Inducible Factor-1 ............. 16 2-1. Tumor hypoxia ......................... 16 2-2. HIF-1 and Cancer ..................... 17 3. Deguelin and SH-1242 ................... 23 MATERIALS AND METHODS 1. Cell culture and hypoxia condition....... 26 2. Antibodies ............................. 27 3. Cell Treatments ......................... 27 4. MTT assay ................................28 5. Western blot .............................28 6. Preparation of conditioned medium ........29 7. Tube formation assay ..................30 8. Flow cytometry analysis...................30 9. Clonogenic growth assay...................31 10. Soft agar colony formation assay.......... 31 11. Generation of Xenograft tumors and Immunofluo rcence.................................... 32 12. Statistical analysis .................... 33 RESULTS 1. SH-1242 has cytotoxicity against NSCLCs, but less than deguelin does in normal bronchial epithelial cells............................. 34 2. SH-1242 exhibits inhibitory effects on colony formation in a dose dependent manner ........36 3. SH-1242 induces apoptosis ..............38 4. SH-1242 inhibits HIF-1ɑ stability and angiogenic activites.....................................40 5. SH-1242 inhibits HSP90 client protein expression ............................................. 43 6. SH-1242 has apoptotic and angiogenic activites in vivo...................................... 45 DISCUSSION.....................................48 REFERENCES.....................................50 ABSTRACT IN KOREAN.............................58Maste

Connectionism and Compositional Structure

천구백팔십년대 초반까지 인지 현상과 마음의 개념에 대한 이론은 현대적인 디지탈 컴퓨터 모형에 근거한 규칙 지배적 기호 조작(rule based symbol manipulation)이론 - 앞으로 고전주의 이론이라고 부르게 될 -이 지배해 왔다. 근 사반세기의 지배였다. 더 정확히 말하자면 천구백사십년대에 이미 이 모형과 견줄 수 있는 모형, 곧 신경망 이론에 바탕을 둔 형식 모형 -연결주의 모형-이 제시되었찌만 그 모형이 갖는 개념상의 한계 때문에 그 동안 경쟁 상대가 되지 못했다. 고전주의는 '계산주의'란 다른 이름에서 알 수 있듯이 인간의 마음을 뉴엘(1980)이 부른 것처럼 물리적인 기호 체계로 본다. 마음이란 본질적으로 기호를 조작하는 정보 처리 시스템이고 그 처리 과정을 일종의 계산 과정으로 이해하고 있는 것이다. 디지탈 컴퓨터 자체의 폭발적인 발전에 힘입어 규칙을 따르는 프로그램을 써서 문제를 해결한다는 모형은 모든 인지에도 적용되어서, 디지탈 컴퓨터가 인간의 마음을 원리적으로 흉내낼 수 있다는 생각이 마음의 본성과 과정, 작용에 궁금해하는 인지과학자들의 관심을 끌게 되었다

Study toward synthesis of saulatine

Maste