• Restorative Dentistry and Endodontics
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• v.29 no.2
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• pp.141-146
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• 2004

The purpose of this study was to evaluate the efficiency of the preparation of oval canals using hand and engine-driven instruments with SEM observation. Thirty single-rooted teeth with oval canal were used in this study. The teeth were divided into 3 groups. In group A the teeth were instrumented up to a size 35 K-file using RC-prep and irrigated with 5% NaOCl between each file size. In group B. the teeth were instrumented with Profile according to the manufacture's instructions using RC-Prep and irrigated with 5% NaOCl between each file size. In group C. the teeth were instrumented with GT file according to the manufacture's instructions using RC-prep and irrigated with 5% NaOCl between each file size. Then. in all teeth. a final flush of 5ml of distilled water delivered for 30s. Canals were dried with sterile standardized paper points. After preparing the canals, the teeth were sectioned along their mesial and diatal surfaces by using low-speed diamond disc. chisel and mallet. Each root section was then dehydrated in graded concentration of alcohol (70, 80, 90, 100%). mounted on an aluminum stub. sputter-coated with gold-palladium and observed with scanning electron microscope (HITACHI S-4200) in middle and apical area. The results of this study were as follows: 1. In the middle area. group B and group C showed less smear layer than group A and it was statistically significant (p < 0.05). 2. In the middle area. group B showed greater smear layer than group C. but it was not statistically significant (p > 0.05). 3. In the apical area, group C showed less smear layer than group A. and it was statistically significant (p < 0.05). 4. In the apical area. group A showed greater smear layer than group B. but it was not statistically significant (p > 0.05). 5. In the apical area. group B showed greater smear layer than group C. but it was not statistically significant (p > 0.05). 6. In all groups. the middle area was less smear layer than the apical area. and it was statistically significant (p < 0.05).

• Parasites, Hosts and Diseases
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• v.34 no.3
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• pp.169-176
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• 1996

A scanning electron microscopic study was performed to observe the surface ultrastructures of the third-stage larvae of Gnathostoma hispidun. The early third-stage larvae (EL3) were collected from the viscera of Chinese loaches by the artificial digestion method . The advanced third-stage larvae (AdL3) were recovered from mice experimentally infected with EL3. Both larval worms were fixed with 2.5% glutaraldehyde, dehydrated in graded alcohol. dryad in critical point dryer, and coated with gold. The specimens were observed with a SEM (DS- l30C). On the head bulb of both larval stage, the mouth had a pair of lateral lips of equal size and of half moon shape. Each lip had a couple of labial papillae and a small amphid located between the two papillae. The hooklets on the head bulb had single-pointed tips and curved posteriorly. The cuticular spines of EL3 were larger and more densely distributed in the anterior area (about 1.8 Mm in length) and gradually decreased in size and number posteriorly. The cuticular spines in the anterior area of AdL3 were sharp-pointed and about 4.5 Mm in length, and those in the middle area were about 1.75 Mm. The velvety cuticular folds and dot-like cuticular spines were distributed in the posterior area. A cervical papilla was located between the 7th and 8th transverse striations. A dome-like body papilla was located at the posterior 1/4 of body. An ellipsoidal excretory pore was located between the 17th and 18th striations. From the above results, it is suggested that the characteristic SEM findings obtained from this study may be helpful on the species identification of larval Gncthostomn.

• Applied Microscopy
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• v.7 no.1
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.