• Title/Summary/Keyword: radioautography

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Cytological Studies of Diapause in the Silkworm, Bombyx mori L. (가잠의 휴면성에 관한 세포학적 연구)

  • Park, Kwang E.
    • Journal of Sericultural and Entomological Science
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    • v.18 no.2
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    • pp.1-60
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    • 1976
  • In present study, radioautography and electron-microscopy were applied by the author to the elucidation of the mechanism of diapause in Bombyx mori L. (1). Patterns of nucleic acid and protein syntheses during embryonic development of silkworms, incubated at high and at low temperatures, were demonstrated by means of radioautography with labelled precursors of nucleic acid and protein. On the third day after blastokinesis the embryo incubated at high temperature generally incorporated much of the $^3$H-glycine into the brain and the suboesophageal ganglion, and some into other regions. When incubated at low temperature, no difference in the pattern between the brain, the suboesophageal ganglion and other parts was observed. Radioautography with $^3$H-thymidine revealed no significant difference in DNA synthesis in embryos incubated at high and low temperatures. In diapausing eggs twenty days after deposition, only a few cells of the mesoderm incorporated the labelled material into their nuclei, but in the hibernated eggs all the nuclei of the mesoderm had thymidine incorporation. After blastokinesis only the anterior portion of the embryo around the brain and the suboesophageal ganglion had thymidine incorporation; this was not observed in the posterior portion.

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RADIOAUTOGRAPHIC AND HISTOLOGIC INVESTIGATION OF SKIN IN YOUNG AND OLD FROGS

  • KOO KOOK BON;You Dong Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.13 no.1
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    • pp.75-85
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    • 1983
  • Age differences in the skin structure have been studied in young (one year-old) and aged (five and a half year-old) frogs, Xenopus laevis. The epidermis in young frogs is made up of an average of 6.3 and 4.7 layers of epithelial cells at abdominal and dorsal surfaces, respectively. In aged frogs, the number of respective cell layers at abdominal and dorsal surfaces increases to 8.8 and 5.6. The thickness of the dermis (spongiosum) in aged frogs is decreased 25% on the abdominal side (from 267㎛ to 207㎛) but is increased by 11 % on the dorsal side (from 275㎛ to 305㎛). The nucleolar index and ³H-uridine incorporation, as judged by radioautography, by epithelial cells are drastically reduced in aged frogs.

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The Study of Iodine Metabolism IN VIVO Utilizing I-131 (방사선 동위원소 I-131을 이용한 요드의 IN VIVO 대사 연구)

  • Byun, Si-Myung
    • Applied Biological Chemistry
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    • v.19 no.2
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    • pp.70-74
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    • 1976
  • In order to study the mechanism of biosynthesis of thyroid hormones, radioactive iodine was injected into the rats and thyroid glands were removed. Iodine compounds hydrolyzed by pancreatin viokase were separated by paper chromatography and analyzed by radioautography. Radioautograms showed that the uptake of iodine starts immediately and forms diiodotyrosine through monoiodotyrosine. Evidence supported the possibility that diiodotyrosine is a precursor of thyrosine and triiodothyronine is a degradation product of thyroxine. The rat administered propylthiouracil showed inorganic iodine concentration activity, while the binding activity was prevented.

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An Electron Microscopic Radioautographic Study of the Synthesis and Migration of the Glycoproteins in the Osteoclast of the Mice Maxillary Alveolar Bone (생쥐 상악치조부에서의 파골세포의 당단백 합성 및 이동에 관한 전자현미경 자기방사법적 연구)

  • Kim, Myung-Kook
    • Applied Microscopy
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    • v.22 no.2
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    • pp.118-126
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    • 1992
  • The pathway and time course of fucose-containing glycoprotein synthesis and intracellular translocation in osteoclasts of the mice maxillary alveolar bone were investigated by electron microscopic radioautography. Male Balb-C mice weighing 17gm were anesthetized with Nembutal and injected via the external jugular vein with 2.5 mCi of $L-[6-^{3}H]-fucose$ (specific activity 16.8 mCi/mmol) in 0.1 ml of sterile saline solution. At 5, 10, 20, 35 minutes and 8 hours after administration of the $^{3}H-fucose$, animals were killed by intracardiac perfusion of 30ml of 2% glutaraldehyde in a modified Tyroid solution, pH 7.4. The maxillae were then removed and further fixed in Karnovsky fixative for an additional 3-4 hours. After rinsing in 0.1M cacodylate buffer for 10 minutes, the maxillae were demineralized for 2 weeks at $4^{\circ}C$ in ethylene diamine tetra acetate containing 2% glutaraldehyde. The first interdental areas were mesiodistally sectioned into slices of 1mm thickness and postfixed in osmium tetroxide. Tissues were then dehydrated and embedded in Poly Bed. To prepare electron microscopic radioautography, the dipping method of Kopriwa (1973) was employed. Thin sections were coated with a crystalline monolayer of ILford $L_4$ photographic emulsion. After exposure for 4 months at $4^{\circ}C$, the sections were developed Kodak Microdol-X and Phenidon (for compact grains), fixed in 30% sodium thiosulfate, stained with uranyl acetate and lead citrate and examined in the electron microscope (JEOL 1200 EX). At 5, 10 and 20 minutes after injection, $^{3}H-fucose$ was concentrated in Golgi cisternae of the osteoblasts. By 35 minutes the labels were observed over small vesicles in the suprannclear area of osteoclasts. At 8 hours, numerous silver grains were located on the ruffled border and cell membrane of osteoclasts. These results indicate that fucose molecules are added in the Golgi apparatus and small vesicles appear to be responsible for translocation of the glycoproteins to the marginal portion of osteoblasts. The glycoproteins are distributed on the osteoclast cell surface and especially over the ruffled border.

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Radioautographical observations of development and appearance of glia cells in brain II. Division and migration of ectodermal glial cell in the brain (뇌신경교세포(腦神經膠細胞) 집단(集團)의 발생(發生)과 이동(移動)에 대한 방사선(放射線) 자기법적(自記法的) 관찰 II. 뇌(腦) 외배엽성(外胚葉性) 신경교세포(神經膠細胞)의 분열(分裂)과 이동(移動)에 대하여)

  • Kwak, Soo-dong
    • Korean Journal of Veterinary Research
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    • v.32 no.4
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    • pp.489-496
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    • 1992
  • The division, distribution and migration of the macroglial cells in the juvenile mouse brain were investigated with the radioautography. Forty mice (ICR) were randomly subdivided into two groups. The twenty mice from group 1 were weighing initially 5 to 6g, aged 10 to 12 days and were sacrificied at 2 hrs, 2, 3, 5, 7, 10, 15 and 20 days after a single intraperitoneal injection of $^3H$-thymichine ($4{\mu}$ Ci/g of body weight). Twenty mice from group 2 were weighing intially 2.5 to 5g, aged 3 to 8 days and were sacrificed at 2 hrs, 2, 3, 5. 7, 10, 15 and 20 days after a single($4{\mu}$ Ci/g of body weight) and/or after intraperitoneal repeated injections($2{\mu}$ Ci/g of body weight/interval) at 2, 3 and 5 days after the first injection. The brain preparations were processed for autoradiogrouphy using Kodak NTB-3 emulsion following development in Kodak D-19, fixation in Kodak fixer, and then stained with cresyl echt violet or hematoxylin counterstain. The labeling index of the ectodermal glial cells in the subependymal layers of the lateral ventricles (SLLV), corpus callosum (CC), molecular layer of the neocortex (MLN ), inner layer except the molecular layer in the neocortex (ILN) and medulla of the cerebrum (MC) were invested. 1. Labeling cells appeared from 2 hour and some of them sustained in the 20 day after injection. In the single injection group, the peak of the labeling index reached a 7.6% at 3 day, 3.6% at 7 day, 3.3% at 2 day, 5.0% at 3 day and 2.3% at 2 day from the SLLV. CC, MLN, ILN and MC, respectively. In the repeated injecton group, the peak of the labeling index reached a 32.0 at 7 day, 11.0% at 10 day, 89% at 7 day, 16.0% at 10 day and 10.8% at 15 day from the SLLV, CC, MLN, ILM and MC, respectively. 2 The glial cells of the SLLV were recognized as to be migrated into the CC and to be not or less to be into the MC and ILN but to be not into the MLN. Glial cell aggregates in the neocotex and MC were recognized as to be proliferated and then disappeared in the itself regions.

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AN ELECTRON MICROSCOPIC RADIOAUTOGRAPHIC STUDY OF PROTEIN SYNTHESIS IN VITRO IN THE PALATAL MUCOSA OF THE RAT

  • KIM Hyun Joo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.17 no.1
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    • pp.51-87
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    • 1987
  • The incorporation of ³H-proline by epithelial and connective tissue elements of rat palatal mucosae was studied in order to investigate the relative levels of protein synthesis by the epithelium and underlying connective tissue cells. Following a sixty minutes incorporation of the radioactive tracer in vitro, it was found that the suprabasal cells had most grains per unit area. Furthermore, the grains were more concentrated over the cytoplasm than the nucleus. This was in contrast with the labeling of basal cells which had twice as many grains over the nucleoplasm than that over the cytoplasm. In intermediate cells; i.e., the spinous layer, the number of silver grains per unit area was decreased from that of the suprabasal cells. In areas where desmosomes were more prominent, many grains were in touch with such desmosomes. However, the labeling appeared to be reduced as soon as the cells became flattened. Moreover, the epidermal keratohyalin granules were relatively free of grains. Except for certain intercellular surfaces the keratinized cells were generally free of grains. On the connective tissue side, silver grains were primarily localized over the fibroblasts with occasional grains being found over palatal muscle cells, neural elements and so on. Most grains over collagenous fibers were found in relation to mature collagen fibrils. Thus, protein synthesis in isolated mucosae of the rat palate appeared to take place both in epithelial and connective elements. There were no apparent tissue alterations caused by the in vitro incorporation procedure utilized under conditions of this study.

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AN ELECTRON MICROSCOPIC, RADIOAUTOGRAPHIC STUDY OF ERYTHROPOIESIS IN VITRO

  • MYUNG No Chul
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.17 no.1
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    • pp.27-49
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    • 1987
  • Using ³H-proline as a radioactive tracer, the relationship between the ultrastructural differentiation and the site of protein synthesis has been investigated in developing red blood corpuscles. The general ultra-structure of erythropoietic cells in differentiation after 60 minutes of in vitro labeling has confirmed the results from previous investigations by Bessis, M., Thiery, J. and others. In dividing nuclei more than two-thirds of the labeling were present at the interface between heterochromatin and euchromatin. In less differentiated cells most of the grains in interphase cells was localized over the nucleus. As the cells continued to develope beyond a stage where cytoplasmic density was clearly increased over other cell lines in bone marrow, the majority of grains localized over the cytoplasmic area was decreased in more mature cells, as judged by the density of cytoplasm, and the structural changes in mitochondria, Golgi complex and polysomal configurations. These results show; 1) that the cytoplasm of erythroblast series does not change under in vitro conditions employed in the study; 2) that protein synthesis in the nucleus occurs largely at the interface between euchromatin and heterochromatin in active nuclei; and 3) that cytoplasmic synthesis of proteins continues to take place well into the normoblast stage solong as the physically visible polysomes are present in maturing red blood corpuscles.

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Enzymatic Radioiodination of Insulin for Radioimmunoassay Use

  • Awh, Ok-Doo;Kim, Jae-Rok
    • Nuclear Engineering and Technology
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    • v.12 no.2
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    • pp.81-87
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    • 1980
  • Insulin was labelled with $^{125}$ I using lactoperoxidase as an oxidizing agent. The reaction product was purified via two stages: a starch gel electrophoresis(SGE) and a Sephadex gel filtration (SF). Upon comparison of the labelling yields and the bindabilities of the labelled insulin to its antibody, it has been found that the enzyme method shows higher yields (50%) and the better bindability to its antibody than the conventional chloramine-T method (35%). By checking the insulin blank labelling mixture with a SGE, a paper chromatography, and a radioautography technique, a by-product in the lactoperoxidase method has been identified. The separated fractions in SGE and SF were also analyzed and discussed.

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