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Ultrastructure of the Testis and Germ Cell Development During Spermatogenesis in Male Crassostrea gigas (Bivalvia: Ostreidae) in Western Korea

  • Kim, Jin-Hee;Chung, Ee-Yung;Choi, Ki-Ho;Lee, Ki-Young;Choi, Moon-Sul
    • The Korean Journal of Malacology
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    • v.26 no.3
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    • pp.235-244
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    • 2010
  • Ultrastructural characteristics of the testis and spermatogenesis of Crassostrea gigas were investigated by Transmission and Scanning Electron microscope observations. The testis is a diffuse organ consisting of branching acini containing differentiating germ cells in a variety of stages. The acinus is surrounded by an intermitent layer of myoepithelial cells andis divided into subcompartments that are partially separated by pleomorphic accessory cells which remain in close contact with germ cells until late stages of development. these accessory cells contain a large quantity of glycogen particles and lipid droplets in the cytoplasm. Therefore, it is assumed that they are involved in the supplying of the nutrients for germ cell development, while any phenomena associated with phagocytosis of undischarged, residual sperms by lysosomes could be find in the cytoplasm of the accessory cells. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves. Mature spermatozoa consist of broad, cap-shaped acrosomal vesicle, subacrosomal material (containing axial rod embedded in a granular matrix), a oval nucleus showing deeply invaginated anteriorly, two triplet substructure centrioles surrounded by four spherical mitochondria, and satelite fibres appear to the distal centriole and plasma membrane. Spermatozoa of C. gigas resemble to those of other investigated ostreids. In particular, the anterior region of the acrosomal vesicle is transversely banded. It is assumed that differences in this acrosomal substructure are associated with the inability of fertilization between the genus Crassostrea and other genus species in Ostreidae. Therefore, we can use sperm morphology in the resolution of taxonomic relationships within the Ostreidea. The spermatozoon is approximately $42-47{\mu}m$ in length including an oval sperm nucleus (about $0.91{\mu}m$ in length), an acrosome (about $0.42{\mu}m$ in length) and tail flagellum ($40-45{\mu}m$). The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9 + 2 structure. These morphological charateristics of acrosomal vesicle belong to the family Ostreidae in the subclass Pteriomorphia.

Effect of Additives on Paper Aging (종이 첨가제가 종이의 노화에 미치는 영향)

  • 윤병호;이명구;최경화
    • Journal of Korea Foresty Energy
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    • v.21 no.2
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    • pp.25-33
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    • 2002
  • One of the critical problems to preserve books and documents in libraries and archives is the deterioration. Some of previous results showed that the major cause of paper deterioration was the acid-catalyzed hydrolysis of the cellulose in paper fibres and aging rate of acidic paper was faster than that of alkaline paper. Therefore, It is necessary to remove the acid in the paper for reducing the rate of paper deterioration. It has been reported to extend the useful life of acidic paper by three to five times. Recently, It has been recognized the need for an effective method of deacidifying large quantities of books and document. However, in the previous many reports little attention was paid to the effect of paper additives. In this paper, We carried out experiment about the effect of additives on paper aging and the effect of deacidification by the gaseous ethanolamines (monoehtanolamine, diethanolamine, triehtanolamine). In result, it was found that the strength of aging was in the order of the alum+rosin>alum >AKD> control and the rate of deacidification was in the order of the monoethanolamine>diethanolamine>triethanolamine. The treatment with the gaseous ethanolamines caused decreasing of brightness and dropping of fold endurances. However, deacidification by combination treatment of the various gaseous ehtnaolamines prevented from decreasing of brightness and dropping of folding endurances.

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Transformation of Asbestos-Containing Slate Using Exothermic Reaction Catalysts and Heat Treatment (발열반응 촉매제와 열처리를 이용한 석면함유 슬레이트의 무해화 연구)

  • Yoon, Sungjun;Jeong, Hyeonyi;Park, Byungno;Kim, Yongun;Kim, Hyesu;Park, Jaebong;Roh, Yul
    • Economic and Environmental Geology
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    • v.52 no.6
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    • pp.627-635
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    • 2019
  • Cement-asbestos slate is the main asbestos containing material. It is a product made by combining 10~20% of asbestos and cement components. Man- and weathering-induced degradation of the cement-asbestos slates makes them a source of dispersion of asbestos fibres and represents a priority cause of concern. When the asbestos enters the human body, it causes cellular damage or deformation, and is not discharged well in vitro, and has been proven to cause diseases such as lung cancer, asbestos, malignant mesothelioma and pleural thickening. The International Agency for Research on Cancer (IARC) has designated asbestos as a group 1 carcinogen. Currently, most of these slats are disposed in a designated landfill, but the landfill capacity is approaching its limit, and there is a potential risk of exposure to the external environment even if it is land-filled. Therefore, this study aimed to exam the possibility of detoxification of asbestos-containing slate by using exothermic reaction and heat treatment. Cement-asbestos slate from the asbestos removal site was used for this experiment. Exothermic catalysts such as calcium chloride(CaCl2), magnesium chloride(MgCl2), sodium hydroxide(NaOH), sodium silicate(Na2SiO3), kaolin[Al2Si2O5(OH)4)], and talc[Mg3Si4O10(OH)2] were used. Six catalysts were applied to the cement-asbestos slate, respectively and then analyzed using TG-DTA. Based on the TG-DTA results, the heat treatment temperature for cement-asbestos slate transformation was determined at 750℃. XRD, SEM-EDS and TEM-EDS analyses were performed on the samples after the six catalysts applied to the slate and heat-treated at 750℃ for 2 hours. It was confirmed that chrysotile[Mg3Si2O5(OH5)] in the cement-asbestos slate was transformed into forsterite (Mg2SiO4) by catalysts and heat treatment. In addition, the change in the shape of minerals was observed by applying a physical force to the slate and the heat treated slate after coating catalysts. As a result, the chrysotile in the cement-asbestos slate maintained fibrous form, but the cement-asbestos slate after heat treatment of applying catalyst was broken into non-fibrous form. Therefore, this study shows the possibility to safely verify the complete transformation of asbestos minerals in this catalyst- and temperature-induced process.

Experimental Studies on Cardiac Catecholamine Content (Catecholamines에 관(關)하여 -제삼편(第三編) 심장(心臟) Catecholamines에 관(關)한 실험적(實驗的) 연구(硏究)-)

  • Lee, Woo-Choo
    • The Korean Journal of Pharmacology
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    • v.9 no.1
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    • pp.1-21
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    • 1973
  • During the last decade extensive studios on catecholamines have evolved new knowledge in the physiology and biochemistry of adrenergic mechanism. Cardiac muscle, receiving adrenergic fibres from the stellate, cervical and thoracic ganglia, has been repeatedly shown to have a specific capacity to uptake and to store catecholamines. The catecholamine stores in cardiac muscle have also been shown to be important sites for the action of numerous drugs. Under normal condition, a certain level of catecholamines is maintained in the stores and serves as the basis for studying the changes in the catecholamine content of the heart. Because myocardial catecholamines play such important role in the patho-physiology of the heart, it would be interesting to compare the normal level of myocardial catecholamines among various species of animals. An occasional study has dealt with myocardial catecholamines of several species add ages of animals but these have been insufficiently comprehensive to afford a basis for an understanding of the importance of these amines as related to species and ages. The present investigation was undertaken to determine whether or not there is any significance of myocardial catecholamines in the course of the evolution and development of animals. Seasonal changes, sex difference and regional and subcellular distribution of myocardial catecholamines were also examined. The concentration of cardiac catecholamines was determined by the spectrophotofluorometric procedure described by Shore and Olin. The results obtained were summarized as follows: 1. As animals phylogenetically progressed larger amounts of catecholamines were resent in their hearts. A negligibly small amount of catecholamine was present in the hearts of the clam, a non-vertebrate. Among the vertebrates, cold-blooded animals (snake, turtle, frog, eel and fish) had less myocardial catecholamines than warm-blooded animals, of which aves (fowl and duck) had less than mammalia (cat, dog, rabbit, rat, cow and pig). The ratio of norepinephrine to epinephrine also was greater as the animals progress phylogenetically. 2. Examination of the regional distribution of cardiac catecholamines in warm-blooded animals showed that the content of the auricle was generally higher than that of the septum and considerably than that of the ventricle, but the differences of contents among these regions were not so marked. 3. In the embryonic chick, cardiac catecholamines were firstly detected on the 4th day of incubation, the time before the cardiac innervation of sympathetic nerves. The concentrations of these catecholamines increased but not markedly on the 6th day of incubation, soon after the innervation of sympathetic nerves to the heart. The level of the cardiac catecholamines fluctuated throughout the remainder of embryonic development. 4. In newborn rat hearts, a considerable amount of catecholamines was present. With the development of the rats, the concentrations of myocardial catecholamines increased. The ratio of epinephrine and norepinephrine fluctuated within the range of 40 to 60 pervent. However, as development progressed, the percentage of norepinephrine continued to rise, attaining the adult value of $80{\sim}90%$ after $45{\sim}60$ days. In contrast, the total amount of epinephrine remained fairly constant throughout the animal's development. 5. No significant sexual differences were observed in the concentration of myocardial catecholamines in the developing rat. 6. The catecholamines in the rabbit hearts increased during the summer season (from May to August) and maintained a fairly constant level in the other seasons of the year. 7. The subcellular distribution of cardiac catecholamines was examined by differential centrifugation of homogenates of cardiac muscles in rabbits, cats and rats. The catecholamines were found to be present approximately 20% in particles of mitochondrial fraction, 45% in particles of microsomal fraction and 35% in soluble supernatant fraction. The particle containing catecholamines in cardiac muscle appears to be two different sizes.

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Structural Characteristics and Physical Properties of Wild Silk Fibres; Antheraea pernyi and Antheraea yamamai (야잠사의 구조특성 및 물리적 성질)

  • 권해용;박영환
    • Journal of Sericultural and Entomological Science
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    • v.36 no.2
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    • pp.138-146
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    • 1994
  • The structural characteristics of Antheraea yamamai and Antheraea pernyi silk were investigated by using x-ray diffraction method, IR spectroscopy and polarizing microscopy. The amino acid composition, fiber density, thermal decomposition temperature and glass transition temperature were also measured for relating these physical properties to the structure in comparison with those of Bombyx mori silk fiber. There was no significant structural difference between A. yamamai and A. pernyi silk fiber on an examination of x-ray diffraction curve and IR spectrum. Both of these wild silk fibers showed double diffraction peaks at the Bragg angle 2Θ16.7˚ and 20.5˚by x-ray diffraction analysis as well as IR absorption peaks for the bending vibration of specific groups related to ala-ala amino acid sequence. On the other hand, the x-ray diffraction curve and IR spectrum of Bombyx mori silk fiber are different from those of wild silk fibers, indicating different crystal structure as well as amino acid sequences. It showed under the polarizing microscope examination that the birefringence and optical orientation factor of wild silk fibers are much lower than those of B. mori silk. Also, the surface of degummed wild silk fibers was characterized by the longitudinal stripes of microfibrils in the direction of fiber axies. The amino acid composition, which is strongly related to the fine structure and properties, was not significantly different between these two wild silk fibers. However, the alanine content was somewhat less and polar amino acid content more for A. yamamai. As a result of fiber density measurement, the specific gravities of B. mori, A. pernyi and A. yamamai were 1.355~1.356, 1.308~1.311, 1.265~1.301g/㎤ in the order, respectively. The calculated crystallinity(%) was 64% for B. mori and 51~52% for wild silk fibers, which showed same trend by IR method in spite of somewhat higher value. The thermal decomposition behaviour was examined by DSC and TGA, showing that the degradation temperature was in the order of B mori, A. prernyi and A. yamamai at around 350$^{\circ}C$. It was also observed by TGA that the decomposition seems to proceed step by step according to their specific regions in the fiber structure, resulting the difference in their thermal stabilities. The glass transition temperature was turned out to be 220$^{\circ}C$ for B. mori, 240$^{\circ}C$ A. yamamai and 255$^{\circ}C$ A. pernyi by the dynamic mechanical analysis. It is expected that the chemical properties are affected by the dynamic mechanical behavior in accordance with their structural characters.

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Studies on Genetics and Breeding in Rainbow Trout(Oncorhynchus mykiss) VII. Fertilization of Fresh Egg with Co-Preserved Sperm and Ultrastructural Changes (무지개 송어의 유전 육종학적 연구 VII. 동결보존시킨 정자와 신선한 난모세포의 수정 및 미세구조적 변화)

  • PARK Hong-Yang;YOON Jong-Man
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.2
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    • pp.79-92
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    • 1992
  • This study was carried out to develop new techniques useful for cryopreservation, thawing and artificial insemination, and ultrastructural changes of cryopreserved spermatozoa in rainbow trout(Oncorhynchus mykiss) . Two extenders, such as Tyrode solution and Whittingham's $T_6$ solution, were used to preserve rainbow trout sperm in refrigerator $(-20,\;-40\;and\;-70^{\circ}C)$ or liquid nitrogen $%(-196^{\circ})$. Hand-stripped semen was diluted to 1:16 with two extenders, an then the semen were frozen after mixing semen and each extender containing 1M or 1.5M DMSO solution to 1:1. After 60 days cryopreserved semen was thawed in a $13^{\circ}$ water bath, and subsequently centrifugated. After centrifugation at 1,000 rpm for 5 min thawed semen was washed with extenders, and then fertilized with fresh eggs. The results obtained in these experiments were summarized as follows: After cryopreservation, over 75% of spermatozoa were appeared motile and the survival rate was high. Following cryopreservation by the addition of cryoprotectant such as DMSO, methanol and glycerol, the fertilization rate of the thawed spermatozoa appeared over $99\%$ compared with the control having $99\%$ of fertilization rate. There was no difference between the control and experimental groups such as $(-20^{\circ}C\;-40^{\circ}C\;and\;-70^{\circ}C)$ and $-196^{\circ}$ in fertilization rate. Following cryopreservation at $-196^{\circ}$ by the addition of 1M DMSO of cryoprotectant, each fertilization rate following 24 hours and hatching rate following 24 days showed $96\%$ and $8\%$ by the addition of BSA, but showed $98\%\;and\;10%$ by no addition of BSA. Following 2 months of cryopreservation by the addition of 1M DMSO of cryoprotectant, there were $10%$ of hatching rate at $-196^{\circ}\;and\;10\%\;and\;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1M methanol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C,\;and\;28\%,\;at\;-70^{\circ}C$ Following 2 months of cryopreservation by the addition of 1M glycerol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C$, and $33\%,\;at\;-70^{\circ}C$. pollowing 2 months of cryopreservation by the addition of 1.5M DMSO of cryoprotectant, there were $27\%$ of fertilization rate at $-20^{\circ}C,\;an\;36\%\;and \;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1.5M glycerol of cryoprotectant, there were $34\% \;of\;fertilization\;rate\;at\;-20^{\circ}C, \;and\;31\%\;and\;31\%,\;respectively,\;at \;-40^{\circ}C\;and\;-70^{\circ}$. Following 2 months of cryopreservation by the addition of 1.5M methanol of cryoprotectant, there were $28\%$ of fertilization rate at $-20^{\circ}C,\;and\;29\%\;and\;28\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C.$ From 10 days and 15 days following fertilization at $13^{\circ}C\;and\;10^{\circ}C$, respectively, the mortality rate of fertilized ova was markedly increased. The middle piece of spermatozoa had two set of central doublets, nine set of outer coarse fibres, and mitochondrial sheath. Spermatozoa went through morphological changes during storage, e.g. winding of flagella, detachment of the nuclear envelope and the plasma membrane from the nucleus of the sperm head. There were $1\%$ abnormal spermatozoa in fresh sperm and about $15\%$ during storage.

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