• Applied Microscopy
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• 제28권1호
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• pp.107-119
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• 1998

This study was designed to investigate the appearence and the characteristics of the apoptotic cells and the process of the joint cavity formation in mouse knee joint. Fetal mouse knee joints from 15 to 19 days of gestation were used. Paraffin-embedded serial sections, stained with H & E for light microscopic observation, Epon 812 embedded thin sections for electron microscopic observation and Lowicryl HM 20 embedded thin sections for immune-electron microscopic observation were prepared. Monoclonal antibodies to $\beta-tubulin$ and polyclonal antibodies to tissue transglutaminase were used for immune-electron microscopic study. The results obtained were as follows. 1. At 15 days of gestation, blood vessels, which have invaded in the mesenchymal cells, were present in the synovium, to form the joint cavity in the future. 2. At 16 days of gestation, the joint cleft was first appeared and several RBCs were present in the joint cleft. The invasion of blood vessels into the joint cleft was continuing, and apoptotic cells were present in the inner cell layer, adjacent to the joint cleft. Necrotic cells were also present in the outer cell layer; they were present 18 days of gestation, but apoptotic cells did not appear after 17 days of gestation. 3. In the apoptotic cells, transglutaminase were localized around vacuoles and the marginal site of the cytoplasm. 4. In the apoptotic cells, tubulin was around the endoplasmic reticulum and the marginal site of the cytoplasm. In the late stage of apoptotic cells, tubulin was localized diffusely in the cytoplasm. Tubulin was also strongly labeled around in the cytoplasm of the neighboring cell at which the apoptotic body was phagocytosed. Tubulin labeled particles were apparently increased in the seperated apoptotic bodies. On the basis of the above findings, it is proposed that during the development of the mouse knee joint, blood vessel invasion first occurs and then apoptosis and cell necrosis follow it. In the apoptotic cell, present in the synovium of the developing knee joint of the mouse. it is suggested that the redistribution of tubulin is associated with apoptotic process. And transglutaminase overexpressed in the apoptotic cell.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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• pp.145-146
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• 2000

The presence of cross-reacting muramidase in Lactobacillus delbrueckii ssp. bulgaricus ULl2 was shown by using monoclonal antibodies raised against an muramidase-2 of Enterococcus hirae ATCC 9790. The separation of protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western immunoblot confirmed the presence of one cross-reacting band in Enterococcus hirae with an estimated molecular mass of 80 kDa, L. bulgaricus cultured cells harvested after 4 and 12 h were submitted to different autolysin releasing procedures and the liberated products were allowed to cross-react with muramidase-2 antibodies in order to estimate the efficiency of each treatment. Although the cultured cells harvested after 4 h yielded only a slight immune-reaction in Western immunoblots against these enzyme monoclonal antibodies, a strong signal was observed for the cell walls obtained from the same experimental conditions and treated with Triton X-100 surfactant. The same phenomenon was also observed by light fluorescence microscopy. Immune-labelling followed by optical and electron microscopy have shown that the muramidase-2 of L. bulgaricus ULl2 was essentially localized in the innermost part of the cell wall.(omitted)

• BMB Reports
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• 제50권2호
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• pp.55-57
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• 2017

Toll-like receptor 4 (TLR4) together with MD2, one of the key pattern recognition receptors for a pathogen-associated molecular pattern, activates innate immunity by recognizing lipopolysaccharide (LPS) of Gram-negative bacteria. Although LBP and CD14 catalyze LPS transfer to the TLR4/MD2 complex, the detail mechanisms underlying this dynamic LPS transfer remain elusive. Using negative-stain electron microscopy, we visualized the dynamic intermediate complexes during LPS transfer-LBP/LPS micelles and ternary CD14/LBP/LPS micelle complexes. We also reconstituted the entire cascade of LPS transfer to TLR4/MD2 in a total internal reflection fluorescence (TIRF) microscope for a single molecule fluorescence analysis. These analyses reveal longitudinal LBP binding to the surface of LPS micelles and multi-round binding/unbinding of CD14 to single LBP/LPS micelles via key charged residues on LBP and CD14. Finally, we reveal that a single LPS molecule bound to CD14 is transferred to TLR4/MD2 in a TLR4-dependent manner. These discoveries, which clarify the molecular mechanism of dynamic LPS transfer to TLR4/MD2 via LBP and CD14, provide novel insights into the initiation of innate immune responses.

• Nuclear Engineering and Technology
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• 제45권1호
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• pp.67-72
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• 2013

Stress corrosion cracking (SCC) behaviors of Alloy 690 were studied in lead-containing aqueous alkaline solutions using the slow strain rate tension (SSRT) tests in 0.1M and 2.5M NaOH with and without PbO at $315^{\circ}C$. The side and fracture surfaces of the alloy were then examined using scanning electron microscopy after the SSRT test. Microstructure and composition of the surface oxide layer were analyzed by using a field emission transmission electron microscopy, equipped with an energy dispersive X-ray spectroscopy. Even though Alloy 690 was almost immune to SCC in 0.1M NaOH solution, irrespective of PbO addition, the SCC resistance of Alloy 690 decreased in a 2.5M NaOH solution and further decreased by the addition of PbO. Based on thermodynamic stability and solubility of oxide, high Cr of 30wt% in the Alloy 690 is favorable to SCC in mild alkaline and acidic solutions whereas the SCC resistance of high Cr Alloy 690 is weakened drastically in the strong alkaline solution where the oxide is not stable any longer and solubility is too high to form a passive oxide locally.

• Applied Microscopy
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• 제28권4호
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• pp.551-561
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• 1998

The autometallographic method was used to demonstrate the localization of mercury deposits in spleen of mouse. The mercury deposits were identified with the light and electron mocroscope. Mice were treated with methylmercuric chloride in the drinking water (demineralized water) for 40 days. Control and mercury treated groups showed no significant differences in mean body weight and spleen weight per one mouse. Mercury grains were appeared in the germinal center of white pulp consist of a preponderancing lymphocytes, not in red pulp and capsule. At the ultrastructural level, mercury deposits were restricted to lysosomes of macrophage and lymphocyte. Specially, volume in lysosomes of the macrophage was increased. These results suggest that mercury localization in lysosomes is associated with the change of immune activity.

• Parasites, Hosts and Diseases
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• 제32권4호
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• pp.243-248
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• 1994

질편모충 항원 분석의 일환으로 막항원의 분석을 시도하였다 수확 세척된 질편모충 HY-l의 homogenate를 sucrose step-gradient를 이용하여 differential centrifugation하였으며 25%/45%의 sucrose 경계면으로녁터 막분회을 얻었다. 분리된 막분회은 transmission electron microscopy를 통하여 순수 분리되었는지 확인하였고 효소면역 전기영동 이적법(EITB)을 이용하여 항원성을 관찰하였으며 그 결과는 다음과 같았다. 분리된 막분획은 투과전자 현미경상에서 extended sheet나 concentric vesicle의 형태가 거의 균질하게 분포하고 있었으며 막 분획에서 특징적으로 나타나는 trilaminal appearance를 보여 질편모충의 막분획이 순수 분리된 것으로 간주할 수 있었다. 분리된 막분획은 EITB상에 토끼의 항혈청과 반응하였을 때 46, 60, 110, 120, 130 및 150 kfDa에서 항원성이 있는 반응대가 관찰되었으며 N-hydroxysuccinimido-biotic으로 표지하여 분리된 표면항원의 녁획과 비교하였을 때 60 KDa의 항원 분획이 서로 일치하는 것으로 나타났다. 따라서 60 kfDa 의 항원 분획은 료면 항원임을 확신할 수 있었다.

• Applied Microscopy
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• 제36권2호
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• pp.131-140
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• 2006

Neurofascin은 L1CAM의 하나로 신경섬유의 발달과정에서 중요한 역할을 하는 것으로 알려져 있다. 말초신경의 수초형성과 관련된 neurofascin의 역할을 알아볼 목적으로 면역형광염색과 면역전자현미경기법을 이용하여 랫드의 수초좌골신경섬유에서 neurofascin의 분포를 추구하여 다음과 같은 결과를 얻었다. 1.수초형성이 진행됨에 따라 좌골신경섬유에서 neurofascin 분포는 매우 심하게 변화되었다. 2. 수초신경섬유에서 neurofascin은 Ranvier마디에서 약하게 국재하였다. 3. Neurofascin은 수초신경섬유의 paranodal loop, Schmidt-Lantermann incisure, 속축삭사이막, 바깥축삭사이막처럼 Schwann세포의 막이 밀착되지 않은 부위에서도 뚜렷하게 국재하였다. 이상의 연구결과로 neurofascin은 Schwann세포의 마주보는 막사이에 이상적인 간격을 유지하는데 어떤 역할을 하는 것으로 생각되며, 수초층에서 물질이동이 가능하게 하는 것으로 보인다.

• 한국재료학회지
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• 제13권8호
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• pp.550-554
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• 2003

Current nanogold cluster synthesized by chemical routine with 11 or 55 atoms of gold has been widely used for immuno chemistry probe as a form of nanocluster conjugated with biomolecules. It would be an undeveloped region that the 1 nm size of nanogold could be made by materials engineering processing. Therefore, objective of this study is to minimize the size of gold nanocluster as a function of operating temperature and chamber pressure in inert gas condensation (IGC) processing. Evaporation temperature was controlled by input current from 50 A to 65 A. Chamber pressure was controlled by argon gas with a range of 0.05 to 2 torr. The gold nanocluster by IGC was evaluated by X-ray diffraction (XRD) and transmission electron microscopy (TEM). The gold nanocluster for TEM analysis was directly sampled with special in-situ method during the processing. Atomic force microscopy (AFM) was used to observe 3-D nanogold layer surfaces on a slide glass for the following biomolecule conjugation step. The size of gold nanoclusters had a close relationship with the processing condition such as evaporation temperature and chamber pressure. The approximately 1 nm size of nanogold was obtained at the processing condition for 1 torr at $1124 ^{\circ}C$.

• Applied Microscopy
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• 제30권2호
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• pp.173-183
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• 2000

사람 무릎관절 윤활막을 구성하는 윤활세포 중 윤활포식세포(phagocytic synovial cell, type A cell)의 기원에 대한 논의는 형태적으로 큰포식세포의 모습을 하고 있는 단핵포식체 계 (mononuclear phagocyte system)의 한 일부로서 아마도 골수(bone marrow)에서 기원되어졌을 것이라고 알려져 있다. 기능적으로도 LCA, HLA-DR과 Ia 항원에 양성반응을 보여 큰포식세포의 일부로 알려졌으나 아직 연구가 부족한 실정이다. 본 연구는 CD14와 활성화된 큰포식세포의 표지물로 알려진 CD105(endoglin)를 이용하여 윤활포식세포의 세포 내 발현부위를 규명하고, 기능적으로 활성화된 큰포식세포와 포식작용의 역할을 수행하는지 여부를 확인하기 위해 사람의 무릎관절에서 윤활세포들을 냉동초미세박절법을 이용한 면역조직화학 기법으로 CD14와 CB105에 대한 금표지를 하여 투과전자현미경으로 관찰한 결과 다음과 같은 결론을 얻었다. 1. CD14는 윤활포식세포의 과립세포질세망과 세포질및 가장자리, 공포 주변 부위에서 표지 되었으며 공포내에서는 표지 되지 많았다. 2. CD105(endoglin)는 윤활포식세포의 세포막 가장자리와 세포질 돌기를 따라 표지 되었으며 공포 주변 부위에서도 표지 되었으나, 공포 내에서는 표지 되지 많았다. 이상의 결과로 보아 사람 무릎관절 윤활세포층에 위치하는 윤활포식세포는 CD14와 CD105의 항원에 대한 표지를 보이므로 활성화된 큰포식세포나 포식작용의 역할을 수행하는 것으로 생각된다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2935-2941
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• 2012

Biological response modifiers (BRMs) can alter interactions between the immune system and cancer cells to boost, direct, or restore the body's ability to fight disease. Mice with ethylnitrosourea- (ENU) induced leukemia were here used to monitor the therapeutic efficacy of lipopolysaccaride (LPS), Bacillus Calmette Guerin (BCG) and sheep erythrocytes (SRBC). Flow cytometry based CD34+ positivity analysis, clonogenicity, proliferation and ultrastructure studies using scanning electron microscopy (SEM) of stem cells in ENU induced animals with and without BRMs treatment were performed. BRMs improved the stem-stromal relationship structurally and functionally and might have potential for use as an adjunct in human stem cell therapy.