• Title/Summary/Keyword: cytochemical reaction

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Cytochemical and Immunocytochemical Study on the Cellulase Activity in the Digestive Tract of the Land Snail Nesiohelix (동양달팽이 Nesiohelix samarangae 소화관에서의 cellulase 활성에 대한 세포화학적 및 면역세포화학적 연구)

  • 정계헌;이용석;김은정
    • The Korean Journal of Malacology
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    • v.14 no.2
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    • pp.149-159
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    • 1998
  • In order to observe the anticellulolytic localization in the epithelia of the digestive tract such as esophagus, crop, and intestine of a Korean land snail N. samarangae, a cytochemical method and a immunogold labelling method were applied. For the cytochemical study on the cellulase activity, Benedict reaction method applied. And for the immunocytochemical study, the rabbit serum immunoglobuins (IgG) was obtained from the rabbits injected with cellulase which was extracted from body fluid of the snail. The digestive tract tissues of N. samarangae were fixed with 4% paraformaldehyde and 2% OsO4 and embedded in Lowicryl K4M at -40$^{\circ}C$ under UV light (360 nm). The thin sections were loaded on the nickel grids and stained with the serum IgG and protein A-gold complex (particle size: 10 nm). Observations were undertaken with transmission electron microscope (Jeol, JEM-1010). The epithelium of the digestive tract was consisted of five types of cells. In the cytochemical study, the reaction products were found along the periphery of the vacuoles derived from the Bebedict reaction. In the immunocytochemical study, the protein-A gold particles were selectively labelled in Type 1, Type 3 and Type 4 cells in intestinal tissue. membranes of rER, in the surrounding cytoplasm of the rER and secretory granules, and in the apical cytoplasm of the cells. On the material being secreted from the apical cytoplasm was also labelled with the immunogold particles. The all results obtained throughtout present study suggest that the intestinal epithelium of the snail N. samarangae seretes cellulase as one of digestive enzymes.

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Cytochemical Study on the Cellulase Activity in the Digestive System of a Land Snail Helix aspersa

  • Jeong, Kye-Heon;Lee, Yong-Seok
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.539-539
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    • 2000
  • A cytochemical study by the Benedict's reaction method was conducted to find out the cellulase activity in the digestive organs of Helix aspersa. Out of the all digestive organs, the stomach, the intestine, and the digestive gland showed cellulase activities under the transmission electron microscopy.

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Comparison of enzyme cytochemical activities between rosetted cells in peripheral blood of Korean native cattle (한우(韓牛) 말초혈액내(末梢血液內) rosette 형성세포(形成細胞)의 효소화학적(酵素化學的) 염색성(染色性) 비교(比較))

  • Kim, Nyeun-su;Song, Hee-jong;Kim, Sun-jae;Seo, Ye-won
    • Korean Journal of Veterinary Research
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    • v.37 no.1
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    • pp.129-135
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    • 1997
  • Peripheral blood mononuclear cells(PBMNC) of Korean native cattle rosetted with Korean goat erythrocytes(KGRBC) and blood monocytes were evaluated for four cytochemical reactions such as acid phosphatase(ACP), alkaline phosphatase-anti-boby(ALP-Ab), ${\alpha}$-naphthyl butyrate esterase(${\alpha}$-NBE) and peroxidase. The results obtained were as follows; In rosetted cells, the positivities of ACP in E AET-DeX, EA and EAC were 70.3%, 22.4% and 25.2%, those of ${\alpha}$-NB were 27.4%, 44.2% and 79.8%, and those of ALP-Ab were 9.5%, 88.3% and 91.5%, respectively. Whereas, the positivity for Peroxidase in monocytes was 100%. In non-rosetted (remained) cells, the positivities of ACP in E AET+DeX. EA and EAC were 41.4%, 57.2% and 61.9%, those of ${\alpha}$-NB were 38.6%, 16.5% and 18.9% and those of ALP-Ab were 98.2%, 5.3% and 6.3%, in order.

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Lipid and Lipase Distribution on Endosperm Cell of Panax ginseng Seed for the Electron Microscope (전자현미경을 이용한 인삼종자 배유세포내의 지질 및 지질가수분해 효소의 분포)

  • 유성철;노미전
    • Journal of Ginseng Research
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    • v.16 no.2
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    • pp.129-137
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    • 1992
  • This study was carried out to investigate the localization of lipids and lipase activity with lipid staining and cytochemical technique in endosperm cells of Panax ginseng C.A. Meyer seed. In endosperm cells of indehiscent seed, protein bodies facing the umbiliform layer are different in electron density during the various degraded processes. Gradually, protein matrix near the cell wall was lysed and electron lucent inclusions appeared on umbiliform layer. The protein body with high electron density and the spherosome with low electron density were observed in endosperm cells. As a result of lipid staining, electron density of spherosome is more intense than those of the protein matrix within the protein body in endosperm cells of indehiscent seed. Free spherical spherosomes within the umbiliform layer have a high electron density. The spherical spherosomes were more electron densed and were uniform in comparison with the cytoplasmic proteinaceous granules in endosperm cells of seed with red seed coat. The major component of spherosome was determined to be lipid. Lipase activity occurs in the spherosome and near the endosperm cell wall facing the umbiliform layer. Cytochemical reaction products of lipase were observed in the spherosome membrane and in the inner regions of spherosome. After protein bodies were digested, lipase activities were observed in free spherosomes and near the cell wall of endosperm cells. Umbiliform layer composing of fibrillized wall and digested materials of the endosperm cell showed a little lipase reaction products.

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A Cytochemical Study on the Acid Phosphatase of Neurons of Developing Chick Embryo Brain (계배 뇌 발생에 있어서 신경세포의 Acid Phosphatase 변화에 관한 효소화학적 연구)

  • Koh, Ki-Seok;Shin, Chu-Og;Yoo, Chang-Kyu;Choe, Rim-Soon
    • Applied Microscopy
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    • v.18 no.2
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    • pp.119-131
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    • 1988
  • The purpose of this study was to investigate the differentiation and degeneration of neurons in developing chick embryo. The activity of acid phosphatase(ACP) was measured and cytochemical study of ACP and ultrastructural changes were observed in prosencephalon, mesencephalon and rhombencephalon from day 4 to day 19 of incubation. As a result, the activity of ACP of all brain region was tend to increase from day 4 to day 19. On day 13, activities of ACP of mesencephalon and rhombencephalon were increased greatly and activity of ACP was decreased each region on day 17. On electron microscopic examination, the reaction product of ACP were localized at GERL complex, lysosome, Golgi body and vacuoles of neurons. Morphologically, disrupted nuclear envelope, mitochondrial destruction, vacuolization and ribosomal crystalization were observed.

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Effects of Wax Gourd Extracts on Adipocyte Differentiation and Uncoupling Protein Genes(Ucps) Expression in 3T3-Ll Preadipocytes

  • Kang, Keun-Jee;Kwon, So-Young
    • Nutritional Sciences
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    • v.6 no.3
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    • pp.148-154
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    • 2003
  • Although various raw plant materials have been demonstrated to exert anti-obesity effects to a greater or lesser extent in both humans and animals when they are used to supplement the diet, it has not been shown extensively that they influence adipocyte cell differentiation involving lipid metabolic gene expressions. Using a well-established 3T3-L1 preadipocyte differentiation system, we decided to look into molecular and cellular event occurring during adipocyte differentiation when raw plant materials aye included in the process, in an effort to demonstrate the potential use of a screening system to define the functions of traditionally well-known materials. To these ends, the effects of ethanol (EtOH) or EtOH/distilled water (DW) extracts of Wax Gourd were examined using cytochemical and molecular analyses to determine whether components of the extracts modulate adipocyte differentiation of 3T3-Ll preadipocytes in vitro. The cytochemical results demonstrated that EtOH or EtOH/DW extracts did not affect lipid accumulation and cell proliferation, although the degree of lipid accumulation was influenced slightly depending on the extract. EtOH extract was highly effective in apoptotic induction during differentiation of 3T3-Ll preadipocytes (p<0.05). Reverse transcription-polymerase chain reaction (RT-PCR) analysis of lipoprotein lipase (LPL), Uncoupling protein (Ucp) 2, 3 and 4 also showed that while LPL expression was not influenced, Ucp2, 3 and 4 were up regulated in the EtOH extract-treated group and down regulated in the EtOH/DW extract-treated group. These changes in gene expressions suggest that the components in different fractions of Wax Gourd extracts may modulate lipid metabolism by either direct or indirect action. Taking these results together, it was concluded that molecular and cellular analyses of adipocyte differentiation involving lipid metabolic genes should facilitate understanding of cellular events occurring during adipocyte differentiation. Furthermore, the experimental scheme and analytical methods used in this study should provide a screening system for the functional study of raw plant materials in obesity research.

Intracellular Digestion and Endosymbiosis in Amoeba proteus (아메바에 있어서 공생과 세포내소화에 관한 연구)

  • Hah, Jae-Chung
    • The Korean Journal of Zoology
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    • v.22 no.2
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    • pp.67-81
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    • 1979
  • Eelctron microscopic cytochemical methods reveal that acid phosphatase activity appears exclusively in vacuoles surrounding established symbiotes. However, copius amounts of acid phosphatase reaction product are visible between and around some of the degenerating symbiotes in the amebae after treatment of chloramphenicol. It is thought that bacteriostasis by chloramphenicol has served to lost the symbiotic interference to intracellular digestion by the ameba and possibly phodphatase enxymes have been implicated in phagocytosis and intracellular digestion of the symbiotic bacteria.

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Ultrastructural Study on the Parenchymal Cell of Korean Planaria (Dugesia japonica) (한국산 플라나리아(Dugesia japonica Ichikawa et Kawakatsu) 유조직의 미세구조에 관한 연구)

  • 장남섭
    • The Korean Journal of Zoology
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    • v.30 no.1
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    • pp.53-70
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    • 1987
  • The morphological study on the parenchymal cells in the adult planaria performed to observe their cytochemical and ultrastructural characteristics. The results are as follows. Nine types of cells are found in parenchyma. 1. Free parenchymal cell: These cells contain several chromatoid bodies around the nucleus. Heterochromatins are evenly dispersed in large nucleus. These cells are abundant in free ribosomes. 2. Fixed parenchymal cells: These cells have well-developed granular endoplasmic reticulum, mitochondria and Golgi complex but they contain the cytosols exhibiting electron-lucencies. 3. Rhabdite-forming cells: These cells contain the electron-dense rhabdite granules of up to about 0.3 x 0.9 $\mu$m in size. Rhabdite-forming cells have well-developed cell organelles, granular endolplasmic reticulum, mitochondria and Golgy complex. 4. A-type of basophilic granule cells: These cells contain irregularly-shaped granules exhibiting alcianophilia. These granules surrounded by a limited membrane, approximately 1.4 x 0.7 $\mu$m in size, are accumulated in the cytoplasm. 5. C-type of basophilic granule cells: These cells contain electron-dense granules of less than 0.2 $\mu$m in size, which exhibit PAS- positive reaction. This type of granule is also found in the muscle layer of parenchyma. 6. D-type of basophilic granule cells: This type of granule cell occurs only in the parenchyma around reproductive organ. The granules have cytochemical characteristics that they exhibit strongly positive reaction with PAS and weakly eosinophilic property. These electron-dense granules, which are 0.2 to 0.6 $\mu$m in length, have oval shapes. 7. E-type of basophilic granule cells: These cells are found only in the parenchyma around re productive organ. The granules contained in a small number in the cell, exhibit PAS-positive reaction and have an average size of 0. 2pm. 8. Eosinophilic granule cells: These cells contain a large number of eosinophilic granules which have relatively diverse sizes from 0.3 x 0.2 to 0.8 x 0.4 $\mu$m. Most of granules are round or irregularly-shaped and highly electrondense. These cells have an array of well-developed granular endoplasmic reticulum of which cisternae are distened. 9. Transparent granule cells contain electron-lucent granules which exhibit negative reactions with three kinds of cytochemical methods used in this experiment.

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Ascophyllum and Its Symbionts. VII. Three-way Interactions Among Ascophyllum nodosum (Phaeophyceae), Mycophycias ascophylli (Ascomycetes) and Vertebrata lanosa (Rhodophyta)

  • Garbary, David J.;Deckert, Ron J.;Hubbard, Charlene B.
    • ALGAE
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    • v.20 no.4
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    • pp.353-361
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    • 2005
  • Ascophyllum nodosum (L.) Le Jolis has a systemic infection with the ascomycete Mycophycias ascophylli (Cotton) Kohlmeyer and Volkmann-Kohlmeyer with which it establishese a mutualistic symbiosis. In addition, A. nodosum is the host for the obligate red algal epiphyte, Vertebrata lanosa (L.) Christensen. Using light and electron microscopy we describe morphological and cytochemical changes occurring as a consequence of rhizoid penetration of V. lanosa into cortical host tissue. Rhizoids induce localized cell necrosis based on physical damage during rhizoid penetration. Host cells adjacent to the rhizoid selectively undergo a hypersensitive reaction in which they become darkly pigmented and become foci for hyphal development. Light and electron microscopy show that M. ascophylli forms dense hyphal aggregations on the surface of the V. lanosa rhizoid and extensive endophytic hyphal growths in the rhizoid wall. This is the first morphological evidence of an interaction between M. ascophylli and V. lanosa. We speculate that M. ascophylli may be interacting with V. lanosa to limit tissue damage to their shared host. In addition, the fungus provides a potential pathway for the transfer of materials (e.g., nutrients and photosynthate) between the two phototrophs.

Cytochemical Evidence on Seasonal Variation of Peroxidase Activities in Cambial Region of Pinus densiflora, Ginkgo biloba, and Populus alba

  • Wi, Seung-Gon;Lee, Kwang-Ho;Kim, Yoon-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.28 no.4
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    • pp.17-24
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    • 2000
  • The peroxidase activity was localized cytochemically to get an insight into its precise function in lignin biosynthesis. In this work, cerium chloride ($CeCl_3$) was used as a trapping agent for hydrogen peroxide ($H_2O_2$) generated from peroxidase. Seasonal variation of peroxidase activities in cambial region of Populus, Pinus, and Ginkgo was investigated at subcellular levels. Under transmission electron microscopy, electron dense deposits of cerium perhydroxide formed by reaction with $H_2O_2$ were observed in cambium and its immediate derivatives. The staining with $CeCl_3$ in cambium varied with growth seasons. The strongest $H_2O_2$ accumulation, regardless of tree species, appeared in May. Staining pattern of $CeCl_3$ in the cambium of poplar indicated that the production of peroxidase started in March before the opening of buds and reached the highest in May and then declined in August. Ginkgo and Pinus showed relatively late generation of $H_2O_2$ production when compared with Populus. Although Ginkgo and Pinus are classified into gymnosperms, however, the generation of peroxidase production and its duration was different from each other. Little staining appeared in all the tree samples collected in September before falling the leaves.

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