• The Korean Journal of Zoology
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• v.26 no.4
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• pp.271-282
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• 1983

The authors observed the ultrastructure of the pigment cells of the frog, Rana nigromaculata Hallowell, during the hibernation. The specimens from the skin were fixed in 2.5% glutaraldehyde-paraform-aldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 2% osimium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with LKB-ultramicrotome. the ultrathin sections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follows. In hibernating phase, pigment cells of the frog were consisted of the three kinds of chromatophores (xanthophore, iridophore and melanophore) in their dorsal skin. The traits of these cells were as follows. 1. Xanthophores A. Xanthophores were filled with pterinosomes and carotenoid vesicles. Many ribosomes, a few mitochondria and glycogen particles were dispersed in the cytoplasm. B. Pterinosomes were spherical or ellipsoidal in shape. They were divided into 6 types (type I, type II, type III, type IV, type V, type VI pterinosomes) by the their inner structure and especially, type I, type II, type III pterinosomes were well developed.

• Journal of Veterinary Clinics
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• v.25 no.4
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• pp.268-273
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• 2008

For evaluation of immune stimulation effect of lacquer tree extracts, lymphocytes were counted by labeling of FITC-conjugated monoclonal antibody in the pheripheral blood of pigs that fed with a fodder supplemented by lacquer tree extracts. Populations of MHC-II+, CD4+, and CD8+T lymphocytes were increased more than 2% level after 1 week feed supply of the lacquer tree extracts. The increase of those T cells reached at maximum level after 2 weeks in the tested group. B lympyocytes with surface IgM were increased 5% after 1 week feed supply of the lacquer tree extracts, and their numbers reached maximum after 2 weeks in the tested group. For the assessment of cytotoxicity of the lacquer tree extracts, morphological changes were examined on the epithelial cells of small intestine from pigs fed with a fodder supplemented by 0.1 % lacquer tree extracts for 6 weeks (the tested group). Thin-sectioned tissue of small intestine was fixed with glutaraldehyde, then coated with gold particles, and the specimen was examined under scanning electron microscope. The villi on the mucus membrane of jejunum and ileum from the tested pigswere enlarged on the tip and were linked each other.

• Korean Journal of Veterinary Research
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• v.43 no.4
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• pp.541-549
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• 2003

The present study investigated the effects of aging on Leydig cells of Sprague Dawley rats. Rats of 3, 6, 12 and 18 months of age were used. Testes of rat were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in epon-araldite. Using $1{\mu}m$ sections stained with methylene blue, qualitative and quantitative morphological studies were performed. Testis incubations were used to determine luteinizing hormone (LH; 100 ng/ml) stimulated testosterone secretory capacity per testis in vitro. Testosterone levels in the incubation medium, and testosterone and luteinizing hormone levels in serum of these four groups of rats were determined by radioimmunoassay. Morphological studies revealed that Leydig cells were more abundant in the testis interstitium at 6, 12 and 18 months when compared with 3 months. The volumes of Leydig cells per testis was significantly higher, at 6, 12 and 18 months of age than those at 3 months. The number of Leydig cells per testis was doubled at 6, 12 and 18 months of age compared with 3 months. The average volume of a Leydig cell was not significantly different between 3 and 6 months of age, however, at 12 and 18 months a significantly lower value was observed. LH-stimulated testosterone production per testis in vitro was reduced by 45% at 6 months of age compared with 3 months; a further significant reduction was observed at 12 and 18 months. Serum testosterone and LH levels were not significantly different between 3 and 6 months of age but at 12 and 18 months a significantly lower value was observed in both groups for these hormones. These results showed that signs of aging are apparent in Leydig cells of Sprague Dawley rats at 12 months of age.

• Journal of Chest Surgery
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• v.29 no.10
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• pp.1111-1117
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• 1996

Structural deterioration of the bioprosthetic xenograft valves due to primary tissue failure occurs in two modes: from fatigue lesions with tear and wear with or without calcification and from calcification with obstruction. Two groups of consecutive 56 patients with the Hancock porcine ortic valve(HM) and of 1 13 patients with the standard-profile onescu-Shiley bovine pericardial valve(ISM) explanted from mitral position at the time of re-replacement surgery for primary tissue failure at Seoul national University Hospital until 1994, were studied for clinical and pathological features. Their ages at primary implant were 31.9 $\pm$9.2 years In HM and 30.4$\pm$ 12.5 years in ISM. Hemodrnamic dysfunction of the failed mitral bloprostheses were predominantly insufficiency in HM(64.3%) and stenosis in ISM(51.3%)(p<0.001). Pathologic findings of the explanted mitral valves reflected these hemodynamic changes, revealing failure more often from tissue damage(tears and wears) in HM and more often from calcification in ISM(p< 0.001). Explant period(from primary implant to explant) was relatively short in ISM(8.7$\pm$2.6years), compared with the one in HM(10.4 $\pm$2.6 years)(p<0.001). In conclusion, both the Hancock and the lonescu-shiley valves would fail from calcification as well as issue damage. However, while the Hancock porcine valves in mitral position failed more frequently from tissue failure and insufficiency, the standard-profile lonescu-Shiley pericardial valves did from calcification and stenosis, especially in young pAtients . Although the possibility of less occurrence of valve failure from mechanical reasons may be expected with newer generation bloprostheses, it does not seem to Improve durability significantly unless further refinement in antimineralization is achieved. Therefore, clinical use of the glutaraldehyde-treated bioprosthetic valves is, at present, limited to the patients of advanced age groups.

• Journal of Periodontal and Implant Science
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• v.33 no.2
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• pp.193-213
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• 2003

The purpose of this study is to evaluate the phenomenon of attachment and spreading of the cultured rat　calvarial cell inoculated on their surface of different kinds of biodegradable membrane which had been used on tissue regeneration on periodontal defects by using scanning electron microscope. In this experiment 30 Sprague-Dawley male rats (mean BW 150gm) were used to harvest abundant number of cell in the short period. The rats were sacrificed by decapitatioan to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Biodegradable barrier membrane were collected with collagen type, and were divided into 3 different kind of surface such as scattered, polarized and fine-net type as their surface texture. Microcover plate which usually used for cell culture was used as control for smooth surface. All the membrane were seeded with cultured calvarial cell on their surface. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. After the culture as designed time, all the membrane were washed with 0.1 M Phosphate Buffered saline and fuxed with 2.5% Glutaraldehyde. And all specimen were treated with $OsO_4$, and Tannic acid before drying the cell for coating the cell with gold. Scanning Electron Microscope was used to observation. The following results were obtained. I. During the whole period of experiment, the phenomenon of cell attachment and spreading were revealed similar pattern to compare with smooth surface culture plate and ordinary culture dish. 2. The shape of cell attachment and spreading on the surface of barrier membrane were observed no remarked difference pattern between smooth surface culture plate and ordinary culture dish. 3. The cytoplasmic process of cultured calvaria cell extent to the deep portion of barrier membrane like as their own proper shape. 4. There were no remarkable relationships between the degree of cultured cell spreading and surface structure of barrier membrane. 5. Slight starified layer of cultured calvaria cell were observed on the scattered type of resorbable membrane, Conclusively, this study thus suggest that cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of carrier for many cell which could be used as new tissue regeneration, and those tissue engeering technique may become an new method in the approach to the repair of bone defects.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.301-320
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• 1995

The purpose of this study was to evaluate a new biodegradable membrane - atelocollagen as a guided tissue regeneration barrier on the dehiscence defects adjacent to the dental implants. 3 beagle dogs were selected for this study and all the mandibular premolars($P_1,P_2,P_3&P_4$) were extracted. Twelve weeks after the extraction, the edentulous ridges were formed to be placed the titanium plasma-sprayed IMZ implants. Four implant osteotomies were performed on each side of the mandible. The osteotomies were placed facially in the edentulous ridges to approximate an actual dehiscence defect as closely as possible, The standardized dehiscence defects were created 3 mm in width and 4 mm in height by osteotomy. A total 24 implants were placed. e-PTFE, ateloco11agen and $Collatape^{(R)}$ were placed to cover the defects and the one defect served as a control, not covered any membrane. By random selection, three dogs were sacrificed at 2 weeks, 4weeks and 8 weeks after fixation with 3% glutaraldehyde. A week before sacrificing, 8-week dog was infused intravenously with oxy-tetracycline 30mg/kg. The left mandibular blocks were used for full decalcified histologic preparation and the right mandibular blocks were selected for undeca1cified preparation, At 2 weeks, the regenerated bone of e-PTFE and atelocollagen groups appeared to be more dense than other groups and the percentage of bone defect fill was highest for e-PTFE and follwed by ateloco1lagen group. However, the $Collatape^{(R)}$ and control groups showed a little new bone formation. $Collatape^{(R)}$ was almost degraded within 2 weeks. At 4 weeks, the regenerated new bone were much greater and denser than at 2 weeks for e-PTFE and ateloco11agen group. Although a part of atelocollagen bagan to be degraded at the margin and surrounded by foreign body giant cells related to foreign body reaction, it was generally intact and the regenerated new bone was shown much more than at 2 weeks. The amount of new bone in $Collatape^{(R)}$ and control groups at 4 weeks were similar to that of 2 weeks group. At 8 weeks, the regenerated bone was matured and observed along the implant fixture. Direct new bone formation and calcium deposits beneath the e-PTFE were observed. No further bone growth was seen in the $Collatape^{(R)}$ and control groups. In reflected fluoromicrcocopic observation, the osteogenic activity was pronounced between e-PTFE membrane and the old bone. High osteogenic activity was also observed in atelocol1agen group. This study suggested that the ateloco11agen as well as e-PTFE could be used for guided tissue regeneration on dehiscence defects adjacent to the dental implants. But the $Collatape^{(R)}$ was completely resorbed within 2 weeks and was not a suitable membrane for guided bone regeneration.

• Applied Microscopy
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• v.17 no.1
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• pp.47-64
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• 1987

Morphological difference of the renal glomerulus at different age groups have been studied in young (three month-old), adult (twelve month-old) and old (thirty month-old) Fisher strain 344 rats. Pieces of the tissues were taken from renal corticies prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1 M Millonig's phosphate buffer, pH 7.3), following by post-fixation with 1% osmium tetroxide (0.1 M Millonig's phosphate buffer, pH 7.3) and embedded within Araldite. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed under a JEM 100CX electron microscope. The mean thickness of glomerular basal lamina and Bowman's capsule were determined by measuring the thinnest portion of basal lamina, and by taking the average of 50 readings from electron micrographs at different ages. The numerical changes of the slit pores were compared based upon the numbers over the length of 10um of glomerular basal lamina. The results were as follow: 1. The thickness of glomerular basal lamina is increased during aging; 140.4 nm in young rats, 270.0 nm in adult ones, and 437.8 nm in old ones. 2. The thickness of basal lamina of parietal cells of Bowman's capsule is 187.5 nm in young rats, 914.0 nm in adult ones, and 2850.0 nm in old ones. 3. The numbers of the slit pores of basal lamina are reduced during aging, 30.3 slit pores/$10{\mu}m$ in adult ones, and 24.2 slit pores/$10{\mu}m$ in old ones. 4. Accumulation of dense intracytoplasmic filamentous material in the parietal cells of Bowman's capsule is increased in the vicinity of the basal lamina during aging. The proximal tubule-like epithelial cell in Bowman's capsule is observed at one glomerulus in a young rat. 5. The endothelial cells are edematous and form balloon-like structure protruding into capillary lumen in young and old rats. 6. Cytoplasm of the podocyte shows a variety of alteration during aging, such as swelling of mitochondria and of endoplasmic reticulum, and increase of microtubules, microfilaments, lysosomes and lamellated myelin structures, etc. Accumulation of dense intracytoplasmic material in the foot processes is increased in the vicinity of the basal lamina during aging. The podocytic membrane-like structures are seen in young and o]d rats. 7. The mesangial matrices and mesangial cells are increased during aging, and slight swelling of endoplasmic reticulum and Golgi cisternae in young and old rats.

• Applied Microscopy
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• v.17 no.1
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• pp.83-97
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• 1987

In order to discriminate the enteroendocrine cell types in the mucosal epithelium of the normal duodenum of the Korean hedgehog (Erinaceus koreanus). The tissues were fixed in the mixture of 1% paraformaldehyde and 1% glutaraldehyde in phosphate buffer (pH 7.2), and postfixed in 2% osmium tetroxide (phosphate buffer, pH 7.2). They were embedded in Araldite, and the ultrathin sections were made by LKB-V ultratome following the inspection of semithin sections stained with toluidine blue-borax solutions. Ultrathin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100B electron microscope. At least six types of enteroendocrine cells distributed in the mucosal epithelium of the duodenum were identified according to their morphological characteristics mainly based on the size, shape, number and electron density of the secretory granules. Type I cells had moderately developed organelles. The secretory granules were pleomorphic ($370X510nm$), and the granule cores with high electron density were enveloped in limiting membrane and characterized by a narrow halo. Type II cells contained an indented nucleus and well-developed organelles. The secretory granules were round (350 nm) and classified in two kinds by electron density, moderate and high. Both granules were surrounded by limiting membrane and those with high electron density showed often a wide halo. Type III cells had an indented nucleus. The secretory granules with various electron density were round (220 nm) in shape. The granules with high electron density were enveloped in limiting membrane and characterized by a narrow halo, but those with low or moderate electron density had not been observed the limiting membrane. Type IV cells contained an indented nucleus and moderately developed organelles. The secretory granules were round (180 nm) in shape, and the granule cores with high electron density were enveloped in limiting membrane and showed often a wide halo. Type V cells had a large amount of rough endoplasmic reticulum. Secretory granules with low or moderate electron density were round (230 nm) in shape, and surrounded by limiting membrane and showed a narrow halo. Type VI cells contained an oval nucleus and well-developed organelles, especially Golgi complex. The secretory granules with high electron density were round (210 nm) in shape. The granules were enveloped in limiting membrane and showed often a wide halo.

• Polymer(Korea)
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• v.24 no.1
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• pp.82-89
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• 2000

The blood compatibilities of PU/PVA polymer blends with different mixing ratios were evaluated using various methods, such as fibrinogen adsorption, plasma recalcification time, platelet adhesion, whole blood clotting time, and complement activation. In addition, PVA on the surface of the polymer blends was crosslinked by glutaraldehyde to restrain the mobility of PVA molecules for characterizing the effect of PVA in the polymer blends on blood compatibility. The fibrinogen adsorption on the polymer blends decreased with the increase of PVA amount in the polymer blends. The plasma recalcification times of the polymer blends with 10-50 wt% PVA were longer than those of PU, PVA, and polymer blends with higher amount of PVA. The morphological changes and adhesion of platelets on the polymer blends with 30-50 wt% PVA were less than those on the other materials. The blood clotting times and complement activation on the polymer blends with 30-50 wt% PVA were reduced, compared to the other materials. On the other hand, the blood compatibility of the crosslinked polymer blends was relatively decreased, compared to the non-crosslinked ones. According to these experimental results, the blood compatibility of the polymer blends with 30-50 wt% PVA was better than that of the other materials and such a blood compatibility of the polymer blends might be related to the mobility of PVA molecules on the surface.

• Applied Microscopy
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• v.31 no.2
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• pp.185-197
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• 2001

Human gastric adenocarcinomas are fixated with low energy of microwave (LEM) to study fixation effects in level of ultrastructure and antigenicity of the cancer. For the Ag-Ab reactions , the LEM fixated sdenocarcinomas are incorporated with monoclonal mouse anti-human p53 (IgG2b, kappa) and rabbit anti human cerbB-2. The retrieval of antigenicity are easily recognizable in the LEM fixated sections compared with that of frozen sections which show often diffused colour reactions. And the LEM fixation methods have preserved ultrastructures of the adenocarcinoma, but it was often difficult to maintain constancy in fixation effects. For the constancy, LEM was coupled with low concentration of chemical fixatives, such as glutaraldehyde (<1%) and $OsO_4$ (<0.5%). The results were acceptable, but there are tendencies that the adenocarcinoma requisitioned rather weak microwave energy to come into the optimal fixation effects. Therefore , cultured HeLa cells were fixated with lower energy of microwave than that used to the adenocarcinoma. The ultrastructures of the single HeLa cell have been preserved. The results may imply that a different energy levels of microwave are requisitioned in accordance with kinds of cells and tissues for the optimal fixation effects. It is reported and discussed that the fixation methods of LEM used in this work could be applied routinely to conceal a insufficient diffusion rate of chemical fixatives into some kinds of cancer without compromising the ultrastructures as well as to improve antigenic quality of frozen sections.