• Title/Summary/Keyword: cytochemistry

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Cytochemical and Biochemical Characteristics of Cellular Adhesion in Amoeba proteus (Amoeba proteus의 표면흡착에 관한 세포화학 및 생화학적 특성)

  • 안태인;곽인희
    • The Korean Journal of Zoology
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    • v.29 no.3
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    • pp.171-180
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    • 1986
  • The effects of proteases, neuraminidase and EDTA on adhesion of amoebae on the substratum, ultrastructure and biochemical composition of the cell surface were studied by concanavalin A (con A) cytochemistry and SDS PAGE. By con A cytochemistry the glycocalyx of the plasmalemma was easily subdivided into outer filamentous (F) layer and the inner amorphous (A) layer. On treatment with neuraminidase, amoebae attached to the substratum and spreaded better than untreated cells exposing the more con A binding sites in A- and F-layer. When the cells were treated with trypsin or proteinase K, cells stayed unattached for 12 and 48 hr, respectively. Con A binding sites of A layer and all of those glycoproteins were removed by proteinase K. On the other hand, trypsin damaged all of the con A binding sites in both A- and F-layer without significant change in PAS-stained profile of the plasmalemma. Some of the mucopolysaccharides of the cell surface were released by these enzymes and EDTA. When the cells were incubated with monovalent con A they did not attch on the substratum and cytolysed. From these results adhesion of amoebae on the substratum appears to be mediated by the interaction of the glycoproteins and mucopolysaccharides of the A layer.

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Distinction Between Reactive Mesothelial and Carcinoma Cells in Serous Effusions by Mucin- and Immune-cytochemical Panel (점액 및 면역 세포화학적 Panel 염색에 의한 장액성 삼출액내 반응성 중피세포와 암종세포의 감별)

  • Kim, Byung-Heon
    • The Korean Journal of Cytopathology
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    • v.9 no.1
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    • pp.1-14
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    • 1998
  • The cytologic distinction of carcinoma cells from reactive mesothelial cells can be difficult, especially in specimens containing abundant reactive mesotheilal cells and inflammatory cells with scant carcinoma cells. This study evaluates the usefulness of mucin and immunocytochemistry for discrimination between reactive mesotheilal cells and carcinoma cells, and sensitivity and specificity of these stains for the detection of metastatic carcinoma in serous effusions. Immunocytochemical panel including mucin cytochemistry with the periodic acid-Schiff(PAS) reaction after or without diastase digestion was undertaken on 127 serous effusion specimens with histologically confirmed diagnoses. The specimens including cell smears and cell blocks were stained with PAS and antibodies to carcinoembryonic antigen(CEA), epithelial membrane antigen(EMA), cytokeratln(CK), and vimentin. The sensitivities of these stains for metastatic carcinoma(127 cases) were 49%(46/94) in PAS, 48%(60/124) in CEA, 89%(97/109) in EMA, 88%(93/106) in CK, and 25%(20/81) in vimentin. The sensitivities of stains for reactive mesothelial cells(36 cases) were 19%(7/36) in EMA, 78%(28/36) in CK, and 75%(27/36) in vimentin. The PAS and CEA stains were not reacted with all cases of benign reactive serous effusions containing abundant reactive mesothelial cells. The specificities of stains for metastatic carcinoma(127 cases) were 100% in PAS, 100% in CEA, 81% in EMA, 22% in CK, and 25% in vimentin. The optimal combination of stains for use in a panel was PAS and CEA. Combined results from these two stains yielded an advanced sensitivity of 8% in PAS and 4% in CEA for metastatic carcinoma. EMA was also cosiderably useful for identification of carcinoma cells. CK and vimentin were not suitable for distinguishing between reactive mesothelial cells and carcinoma cells.

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Extracellular Triacylglycerol Lipases Secreted by New Isolate of Filamentous Fungus

  • Lusta, Konstantin A.;Woo, Sahng-Young;Chung, Il-Kyung;Sul, Ill-Whan;Park, Hee-Sung;Shin, Dong-Ill
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.832-838
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    • 1999
  • Two different types of lipases (lipase I and lipase II) secreted into culture medium by Rhizopus sp. L-I were purified using a hydrophobic chromatography and were partially characterized. Both enzymes were monomeric as revealed by SDS-PAGE and gel filtration. The molecular masses of the enzymes were identified as 45 kDa (lipase I) and 69 kDa (lipase II). The isoelectric points were estimated to be 3.6 and 5.2 for lipase I and lipase II, respectively. pH and temperature activity optima for lipase I were as 7.5 and $50^{\circ}C$, respectively, whereas the corresponding parameters for lipase II were 6.0 and $45^{\circ}C$. The amino terminal sequences of lipase I and lipase II, determined by Edman degradation, were found to be Leu-Val-Met-Ile-Gln-Arg and Leu-Val-Met-Lys-Gln-Arg, respectively. By western blotting analysis, the two lipases were found to have a common antigenic determinant. Immuno-electron cytochemistry conducted with polyclonal anti-lipase I antibody indicated the enzyme located in both the periplasm and the adjacent vesicles of fungal hyphae. Fortunately, the sites on the cell envelope where lipase was exported into the culture medium was also identified.

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Cytochemistry of cellulase in Capitate Glandular Trichomes of Pelargonium ${\times}$ fragrans (Geraniaceae)

  • Nam, Ko-Kyung;Lee, Kyung-Whan;Lee, Sang-Eun;Kim, Eun-Soo
    • 한국전자현미경학회:학술대회논문집
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    • 2007.05a
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    • pp.75-78
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    • 2007
  • The localization of cellulase were investigated in the capitate glandular trichomes of Pelargonium ${\times}$ fragransby a transmission electron microscopy. The secretory cells of capitate trichomes involved in biosynthesis and its secretion. Secretory material is transported to the space between the plasma membrane and cell wall, and subsequently accumulated in the secretory cavity. The splitting of secretory cell wall during the formation of secretory cavity is suggested that wall-forming enzymes, such as cellulase, may contribute to the wall separation process. Cellulase reaction product was localized in the secretory cell, the secretory cavity and in the subcuticular wall of glandular trichomes. Reaction products were present over fibrillar matrix in the secretory cavity associated with both the inner wall and the subcuticular wall.

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Studies on Ultrastructure and Several Phosphatase Activity in the Non-parenchymal Cell of the Developing Rat Liver (발생과정중 흰쥐 간 Non-parenchymal Cell의 미세구조 및 수종 Phosphatase의 활성에 관하여)

  • Deung, Y.K.;You, K.H.;Chung, M.H.;Seo, Y.H.
    • Applied Microscopy
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    • v.12 no.1
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    • pp.11-21
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    • 1982
  • A number of recent ultrastructural studies have shown marked differences between the two lining cell types in adult liver sinusoids, endothelial cells and Kupffer cells. In the present study, the ultrastructural features and electron microscopic cytochemistry of sinusoidal lining cells in the fetal liver were studied through fetal period to neonate in the rat. At fetal period, the sinusoid, which contains various blood component, in lined by the endothelial cells, the Kupffer cells and the fat storing cells that located in the space of Disse. As gestation proceeded, these eel's are arranged as adult liver sinusoids. The sinusoidal wall appears to be discontinuous with open fenestration between endothelial cells, but no basal lamina can observed. It seems to be morphologically and functionally distinct at the early gestation between the endothelial cells and the Kupffer cells, the latter showing marked phagocytized activity. The fat storing cells, which contain several fat droplets, are located in the space of Disse. Ultrastructural localization of the acid and alkaline phosphatase activity were noted on the sinusoidal lining cells.

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The glyoxysomal nature of microbodies complexed with lipid globules in Botryospheria dothidea.

  • Kim, K.W;Park, E.W.;Kim, K.S.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.119.1-119
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    • 2003
  • The glyoxysomal nature of microbodies was determined in Botryosphaeria dothidea hyphae based on morphology and in situ enzyme characteristics by transmission electron microscopy and cytochemistry. Bound by a single membrane, microbodies had a homogeneous matrix and varied in size ranging from 200 to 400 m in diameter. Microbodies had crystalline inclusion(s) which consisted of parallel arrays of fine tubules in their matrices. Microbodies and lipid globules were frequently placed in close association with each other, forming microbody-lipid globule complexes in hyphae. The cytochemical activities of catalase and malate synthase were localized in matrices of microbodies, showing intense electron-density of the organelle. In addition, the immunogold labeling detected the presence of catalase in multivesicular bodies and hyphal cell walls as well as in matrices and crystalline inclusions of microbodies, supporting the enzyme secretion through cell walls. Meanwhile, isocitrate Iyase was localized only in matrices of microbodies. These results suggest that microbodies, particularly complexed with lipid globules, in the fungal hyphae are functionally defined as glyoxysomes, where glyoxysomal enzymes are biochemically active for the glyoxylate cycle to be a metabolic pathway in gluconeogenesis. (Mycology and Fugus Diseases)

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Ultrastructure and Activity Pattern of Peroxidase in Secretory Trichomes of Drosera capensis (장대끈끈이주걱 분비모의 미세구조와 peroxidase 활성)

  • Kim, Eun-Soo;Oh, Seung-Eun;Yu, Seong-Cheol
    • Applied Microscopy
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    • v.28 no.3
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    • pp.399-414
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    • 1998
  • Glandular trichomes present on the leaf surface of Drosera capensis were examined using transmission electron microscopy. A large number of stalked glands exist on the adaxial surfaces of the leaf blade. The secretory head is composed of two layers of secretory cells, one layer of middle cells, and the inner tracheids. The secretory cells contain rough endoplasmic reticulum, mitochondria, plastids, Golgi apparatus, and vacuoles. The secretory cells show prominent cell wall ingrowth, and thick cuticle restricted on the subcuticular wall. Frequently, the cuticle has some pores, canal-like structures, showing electron -dense granules being penetrated through them. Ultrastructural localization using diaminobenzidine showed the electron-dense deposits in the vacuole. No peroxidase activity was seen in the cell wall and cytolasm. The activity of peroxidase (POX) isozymes in Drosera which isoelectric point (pI) is 3.6 and some anionic POX isozymes which pIs are laid between 3.6 and 4.6 were especially increased according to the development and the formation of glandular trichomes. Also, the activity of some POX isozymes which isoelectric points are laid between 4.6 and 5.1 were increased in the regions of leaves which has trichomes.

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Effect of Occlusion upon Rat Skin on the Activities of Cutaneous Oxygen Radical Metabolizing Enzymes in Rats

  • Han, Sun-Il;Yoon, Chong-Guk;Cho, Hyun-Gug
    • Biomedical Science Letters
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    • v.7 no.2
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    • pp.71-77
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    • 2001
  • To evaluate the effect of occlusive skin on the activity of cutaneous oxygen free radical metabolizing enzymes in rats, the dorsal skin was covered with closed glass chamber shaped petri dish, 46 mm in diameter and 10 mm in height and sealed by an adhesive. Five day-occluded group showed more increased activity of xanthine oxidase (XO) than that of control, and the activity of five day-occluded group was higher than that of ten day-occluded group. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) were significantly higher in ten day-occluded group than in control or five day-occluded group. All the more, five day-occluded group showed the decreasing tendency of SOD and GPx activities compared to those of control. On the other hand, the cerrous perhydroxide deposits were observed in the intercellular space of the stratum basale in five day-occluded group under the electronic microscope using a cytochemistry method. Futhermore, the degree of cerrous perhydroxide reaction was lower in ten day-occluded group than in five day-occluded group. In conclusion, the increased XO activity and the decreased SOD and GPx activities are likely to responsible far the accumulation of $H_2O_2$ in five day-occluded group.

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