• Applied Microscopy
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• v.12 no.2
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• pp.55-68
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• 1982

The study on the nerve cells in the pars intercerebralis(IP) of 5-day-old cabbage butterfly Pieris rapae L. was performed to observe their ultrastructures and classify them on the basis. of the differences in size, shape and relative distribution cf cell organelles. The brain-subesophageal ganglion complex was fixed in 1% paraformaldehyde-1% gluaraldehyde mixture and embedded in araldite mixture. The transverse thin sections of IP were stained with uranyl acetate and lead citrate and examined by Hitachi 500 and ]EM 100B electron microscope. Five distinct types. of nerve cells are recognized and are arbitrarily designated as Type I, Type II Type III, Type IV and Type V. Type I neurone: These neurones are neurosecretory cells. Several neurosecretory cells are. recognized in the pars intercerebralis. They are roughly round or peach-shaped cells measuring $13{\sim}25{\mu}m$ in diameter. The rounded nucleus shows about $5{\sim}10{\mu}m$ in diameter. The chromatin is predominantly diffused with only occasional dense patches. The perikaryon contains numerous. mitochondria, free polyribosomes and neurosecretory granules. The neurosecretory granules are relatively uniform in electron density, and each one is about $100{\sim}400{\mu}m$ in diameter and surrounded by a single membrane. The granules are also observed mostly as in groups. In one group of neurones the cisternae of endoplasmic reticulum are distended or in other group of neurones are not distended. Golgi saccules are slightly dilated at their lateral extremities and contains. frequenty dense rounded materials. Type II neurone: Thes have the largest soma in the pars intercerebralis about $30{\sim}35{\mu}m$ in diameter. They also show roughly polygonal in shape. The nucleus is elongated or sickle-shaped. The chromatin is mainly in the euchromatin form. The perikarya in these cells are well populated with populated with free ribosomes and contain numerous mitochondria and Golgi bodies. The cisternae of granular endoplasmic reticulum are also well distributed. Type III neurone: They are oval or spindle-shaped and also medium-sized. neurones approximately $15{\sim}17{\mu}m$ in length. The nucleus is oval or slightly elongated in shape and $8{\sim}9{\mu}m$ in length. The chromatin occurs in diffused form. The cytoplasm contains many filamentous or oval mitochondria. The perikaryon has also numerous free polyribosomes and cisternae of granular endoplasmic reticulum. Type VI neurone: They are roughly polygonal in shape probably due to the close approximation of the adjacent cells. The soma is about $7{\sim}8{\mu}m$ in diameter. The nucleus is round or oval in shape and $5.0{\sim}5.8{\mu}m$ in diameter. The necleus also occupies a large proprion of the cell body. The perikaryon is well populated with free ribosomes and contains several mitochondria and cistenae of granular endoplasmic reticulum. Type V neurone: These neurones are similar to Type VI neurones in various respects such as cell size and cell inclusion, but they differ from Type IV neurones in shape. The soma is oval or slightly elongated. The cell body contains several filamentous and oval mitochondria.

• Applied Microscopy
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• v.22 no.2
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• pp.1-14
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• 1992

This experiment was performed to study the morphological responses of the parafollicular cells of rat following X-ray irradiation. Male rats were divided into normal and experimental groups. The head and neck region of the rat, under sodium thiopental anesthesia, was exposed to 3,000 rads or 6,000 rads of radiation in a single dose, respectively. The source was a Mitsubishi Linear Accelerator ML-4MV. The target to skin distance was 80 cm, and the dose rate was 200 rads/min. The rate of experimental groups were sacrificed on the 6th hour, 2nd and 6th day after X-ray irradiation. Pieces of the tissue taken from the thyroid gland were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1M Millonig's phosphate buffer, pH 7.3), and in 1% osmium tetroxide (0.1M Millonig's phosphate buffer, pH 7.3), and embedded in araldite mixture. The ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100 CX-II electron microscope. The results were as follow; 1. Two types of the parafollicular cells, according to their electron densities, were found, i. e., light cells and dark cells. 2. Three types of the parafollicular cells, according to their sizes of secretory granules were found, i.e., small granule cells ($85nm{\pm}20.1;64{\sim}102nm$), medium granule cells ($120nm{\pm}26.5;77{\sim}179nm$), and large granule cells ($165nm{\pm}25.7;128{\sim}189nm$). 3. The differential ultrastructural changes of the cells according to their cell types, i.e., dark and light cell, or small, medium and large granule cells, were hardly observed in the time and dose range covered by this study. 4. The morphological changes of the parafollicular cells were not pronounced after exposure to 3,000 rads of X-ray. 5. Swollen cisternae of the granular endoplasmic reticulum and partial cytolysis were observed after exposure to 6,000 rads of X-ray. 6. Above results suggest that the parafollicular cells showed the alterations of mitochondrial and granular endoplasmic reticular swelling, and partial cytolysis, but only in doses of 6,000 rads.

• Applied Microscopy
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• v.22 no.2
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• pp.15-29
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• 1992

The gallbladder is known to have the function of the storage and the concentration of the bile produced by the liver. This function is carried out by the removal of water and inorganic electrolytes. Extrahepatic cholestasis or the impairment of excretion of the bile leads to the distension and loss of the function of the gallbladder. The purpose of this study was to examine the ultrastructural characteristics of the normal gallbladder epithelial cells, and their structural changes induced by the ligation of common bile duct of the rabbit. Common bile duct ligation was performed under ether anesthesia. The rabbits were sacrificed on the 1st, 3rd, 5th, 7th and 14th day, respectively after operations. The tissue blocks of the gallbladder were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde prior to fixation in 1% osmium tetroxide, and embedded in the araldite mixture, and observed with JEM 100 CX-II electron microscope. The results were as follows: 1. The normal gallbladder epithelium of adult rabbit demonstrated two cell types, the ordinary epthelial cell and the dark cell. The dark cells have electron dense cytoplasm, and were found much infrequently, whereas ordinary epthelial cells were found quite numerous. 2. The ordinary epthelial cells of normal gallbladder were provided with the regular microvilli at the free surface and the images of pinocytotic activities in the apical cytoplasm, and exhibit highly convoluted lateral surfaces with elaborated microfolds. These figures of the cells suggest that they are resorptive in functional activity. 3. In the early stages (1st, 3rd, 5th day groups) following the ligation, the apical cytoplasm of some cells is protruding from the free surface and lost their microvilli. Numerous mucous granules filled in the apical and supranuclear cytoplasm compactly. 4. In the late stages (7th, 14th day groups) following the ligation, many light cells containing mumerous mucous granules are seen, between the ordinary epthelial cells. Mucous granules are fused each other, and are discharged into the lumen from the apical cytoplasm. The lateral membranes are straight or undulating without any interdigitations. From the above results, it was concluded that in the cholestasis induced by the common bile duct ligation, there is a tendency for the mucosal epithelium of the rabbit gallbladder to have secretory rather than an absorptive function.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.37 no.3
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• pp.195-204
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• 2011

Introduction: This study examined the regulatory mechanism underlying the meal-induced changes in the hypothalamic-pituitary-adrenal gland (HPA) axis activity. Materials and Methods: Male Sprague-Dawley rats (250-300 g) were hired for two different experiments as follows; 1) rats received either 8% sucrose or 0.2% saccharin ad libitum after 48 h of food deprivation with the gastric fistula closed (real feeding) or opened (sham feeding). 2). rats received 5 ml of intra-oral infusion with 0.2% saccharin or distilled water after 48 h of food deprivation. One hour after food access, all rats were sacrificed by a transcardiac perfusion with 4% paraformaldehyde. The brains were processed for c-Fos immunohistochemistry and the cardiac blood was collected for the plasma corticosterone assay. Results: Real feedings with sucrose or saccharin and sham feeding saccharin but not sucrose, following food deprivation decreased the plasma corticosterone level. c-Fos expression in the nucleus tractus of solitarius (NTS) of the fasted rats was increased by the consumption of sucrose but not saccharin, regardless of the feeding method. On the other hand, the consumption of sucrose or saccharin with real feeding but not the sham, induced c-Fos expression in the paraventricular nucleus (PVN) of the fasted rats. The intra-oral infusion with saccharin or water decreased the plasma corticosterone level of the fasted rats. Intra-oral water infusion increased c-Fos expression in both the PVN and NTS, but saccharin only in the NTS in the fasted rats. Conclusion: Neither restoration of the fasting-induced elevation of plasma corticosterone nor the activation of neurons in the PVN and NTS after refeeding requires the palatability of food or the post-ingestive satiety and caloric load. In addition, neuronal activation in the hypothalamic PVN may not be an implication in the restoration of the fasting-induced elevation of the plasma corticosterone by oropharyngeal stimuli of palatable food.

• Journal of Periodontal and Implant Science
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• v.32 no.4
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• pp.733-744
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• 2002

The fibroblasts are the principal cells in the periodontal ligament of peridontium. As the periodontal ligament fibroblasts (PDLF) show similar phenotype with osteoblasts, the PDLF are thought to play an important role in alveolar bone remodeling. Cell-to-cell contacted signaling is crucial for osteoclast formation. Recently it has been reported that PDLJ enhance the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The aims of this study were to $clarify\;^{1)}$ the mechanism of PDLF-induced osteoclastogenesis $and\;^{2)}$ whether we can use preosteoclast cell line instead of primary hematopoietic preosteoclast cells for studying the mechanism of PDLF-induced osteoclastogenesis. Osteoclastic differentiation of mouse macrophage cell line RAW264.7 was compared with that of mouse bone marrow-derived M-CSF dependent cell (MDBM), a well-known hematopoietic preosteoclast model, by examining, 1) osteoclast-specific gene expression such as calcitonin receptor, M-CSF receptor (c-fms), cathepsin K, receptoractivator nuclear factor kappa B (RANK) ,2) generation of TRAP(+) multinucleated cells (MNCs), and 3) generation of resorption pit on the $OAAS^{TM}$ plate. RAW264.7 cultured in the medium containing of soluble osteoclast differentiation Factor (sODF) showed similar phenotype with MDBM-derived osteoclasts, those are mRNA expression pattern of osteoclast-specific genes, TRAP(+) MNCs generation, and bone resorbing abivity. Formation of resorption pits by osteoclastic MNCs differentiated from sODF-treated RAW264.7, was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for ODF, to the sODF-containing culture me야um. The effects of PDLF on differentiation of RAW264.7 into　the TRAP(+) multinucleated osteoclast-like cells were examined using coculture system. PDLF were fxed with paraformaldehyde, followed by coculture with RAW264.7, which induced formation of TRAP(+) MNCs in the absence of additional treatment of sODF. When compared with untreated and fixed PDLF (fPDLF), IL-1 ${\beta}$-treated, or lipopolysaccha-ride-treated and then fixed PDLF showed two-folld increase in the supporting activity of osteoclastogenesis from RAW264.7 coculture system. There were no TRAP(+) MNCs formation in coculture system of RAW264.7 with PDLF of no fixation. These findigs suggested that we can replace the primary hematopoietic preosteoclasts for RAW264. 7 cell line for studying the mechanism of PDLF-induced osteoclastogenesis, and we hypothesize that PDLF control osteoclastogenesis through ODF expression which might be enhanced by inflammatory signals.

• Applied Microscopy
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• v.37 no.3
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• pp.167-174
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• 2007

Heat shock protein (HSP) 70 functions as a molecular chaperon and reduces stress-induced denaturation and aggregation of intracellular proteins. Erythropoietin (EPO) plays an important role during acute renal failure repair process by rapidly correcting anemia and enhancing renal tubular regeneration. The purpose of this study was to examine the effect of EPO treatment on renal HSP70 expression. Male Sprague-Dawley rats were injected rHUEPO. Kidney were preserved by in vivo perfusion with paraformaldehyde-lysine-periodate (PLP) and processed for immunohistochemistry and electron microscopy. In control kidney, HSP70 was expressed in the cortex, outer medulla and inner medulla. Especially, HSP immunoreactiviy was mainly founded in descending thin limb of outer medulla and inner medullary collecting duct. In EPO treated kidney, HSP70 expression markedly increased in the descending thin limb of outer medulla and newly detected in cortical collecting duct. Electron microscopy showed the presence of HSP immunoreactivity on the intracelluar vesicles and Golgi complex of descending thin limb and cortical collecting duct. These findings suggest that EPO treatment leads to new production of HSP70 in renal tubular cells, and induction of HSP70 by rHuEPO is causally related to protective function.

• Parasites, Hosts and Diseases
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• v.23 no.2
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• pp.269-284
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• 1985

The metabolism of Entamoeba histolytica would be affected by various environmental factors, and alteration of the environment was known to afEect the fine structure of 5. histolytica. The present study was designed electronmicroscopically to investigate the ultrastructure and enzyme activities in the aEonic and conventional strains of 5. histolytica. The trophozoites of axenically cultivated HK-9 strain and conventional YS-27 and YS-49 strains of 5. histolytica were collected and liKed with 4% paraformaldehyde/0.1M cacodylate buffier(pH 74), After washing them by centrifugation, 1% warm agar was added in the sediment. Solidified agar with the trophozoites was cut into $lmm^3$ cubes, and incubated in the various substrates to observe enzyme activities. Then, the specimen was post-fixed with 3% glutaraldehyde/0.1M cacodylate buffer (PH 7.4) and 1% osmium tetroBide/0.1M cacodylate buffier (pH 7.4) , dehydrated in ascending ethanol series and embedded in epoxy resin. These were sectioned on an ultramicrotome and observed with a transmission electronmicroscope. The procedures for the observation of the fine structure were same as the above, except for the incubation in the substrate. The sections were stained with uranyl acetate and lead citrate. For the observation of the surface of the amoebae, scanning-electronmicroscopy was carried out. The results obtained in the present study are summarized as follows: 1. The fuzzy coat around double-layered plasma membrane of 5. histolytica was more irregularly and densely distributed in the conventional strains (YS-27, YS-49 strains) than in the axonic strain (HK-9 strain). 2. The endosomes, button bodies and chromatin material were surrounded by a double-layered nuclear membrane having scattered nuclear fores. The paranuclear body, mono- or double-layered vacuoles, vacuolar membrane whorls, rosette-like cylindrical bodies, aggregation of cylindrical bodies and helical bodies were found in the cytoplasm of the amoebae. Helical bodies and glycogen granules were generally abundant, while a few smooth endoplasmic reticula were observed in the cytoplasm. 3. Alkaline phosphatase activity was mainly demonstrated in the plasma membrane, limiting membranes of vacuoles and smooth endoplasmic reticula. ATPase activity was observed in the nucleus, limiting membranes of vacuoles and vacuolar membrane whorls. 4. Acid phosphatase activity was commonly demonstrated in the limiting membranes an contents of vacuoles, Iysosome-like organelles, plasma membrane and the button bodies in the nucleus. The activity was more weakly demonstrated in the HK-9 strain than in the other conventional strains of 5. histolytica. No peroBidase activity was observed in the amoeba strains employed in the present study. 5. With a scanning electron-microscope, no distinct structural differences were observed between the amoeba strains. All the trophozoite forms of the amoebae showed crater-like depressions and rugged features on the outer surface.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.1-12
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• 2002

The periodontal ligament(PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. PDL-specific protein;PDLs22 had been previously identified as a novel protein isolated from cultured human PDL fibroblasts using subtraction hybridization between human gingival fibroblasts and PDL fibroblasts. The aim of this study was to examine the expression pattern and tissue localization of PDLs22 protein in embryonic and various postnatal stages of developing mouse using immunohistochemical staining. Embryos (E18) and postnatal (P1, P4, P5, P15, P18) were decapitated and the heads were fixed overnight in a freshly prepared solution of 4% paraformaldehyde. Some specimens were decalcified for $2{\sim}4$ weeks in a solution containing 10% of the disodium salt of ethylenediamine-tetraacetic acid (EDTA). Next, tissues were dehydrated, embedded in paraffin and sectioned serially at $6{\mu}m$ in thickness. Polyclonal antiserum raised against PDLs22 peptides, ISNKYLVKRQSRD, were made. The localization of PDLs22 in tissues was detected by polyclonal antibody against PDLs22 by means of immunohistochemical staining. The results were as follows; 1. Expression of PDLs22 protein was not detected in the tooth germ of bud and cap stage. 2. At the late bell stage and root formation stage, strong expression of PDLs22 protein was observed in developing tooth follicle, osteoblast-like cells, and subodontoblastic cells in the tooth pulp, but not in gingival fibroblasts, ameloblasts and odontoblasts of tooth germ 3. In erupted tooth, PDLs22 protein was intensely expressed in PDL and osteoblast-like cells of alveolar bone, but not in gingival fibroblasts, mature osteocytes and adjacent salivary glands. 4. In the developing alveolar bone and mid-palatal suture, expression of PDLs22 protein was seen in undifferentiated mesenchymal cells and osteoblast-like cells of developing mid-palatal suture, but not in mature osteocytes and chondrocytes. These results suggest that PDLs22 protein may play an important role in the differentiation of undifferentiated mesenchymal cells in the bone marrow and PDL cells, which can differentiate into multiple cell types including osteoblasts, cementoblasts, and PDL fibroblasts. However, more researches should be performed to gain a better understanding of the exact function of PDLs22 protein which related to the PDL cell differentiation.

• Journal of Periodontal and Implant Science
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• v.35 no.4
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• pp.939-948
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• 2005

The role of the periosteum on osteointegration of $Bio-Oss^{(R)}$(Geistlich, Wolhusen/Switzerland) was studied in rabbit calvarial defect. 12 New Zealand white male rabbits between 2.8 and 4 kg were included in this randomized, blinded, prospective study. Each rabbit was anesthetized with Ketamine HCl(5 mg/kg) and Xylazine HCl(1.5 ml/kg). An incision was made to the bony cranium and the periosteum was reflected. Using a 6-mm trephine bur(3i. USA), four 8-mm defects were created with copious irrigation. The defects were classified into barrier membrane($Tefgen^{(R)}$, Lifecore Biomedical. Inc, U.S.A.) only group as a control, $Bio-Oss^{(R)}$ with barrier membrane group, $Bio-Oss^{(R)}$ with periosteum covering group, and $Bio-Oss^{(R)}$ without periosteum covering group. There were 2 rabbits in each group. The wound was closed with resorbable suture materials. Rabbits were sacrificed using phentobarbital(100 mg/kg) intravenously at 1, 2, and 4 weeks after surgery. The samples were fixed in 4% paraformaldehyde, and decalcified in hydrochloric acid decalcifying solution(Fisher Scientific, Tustin, CA) at $4^{\circ}C$ for 2-4 weeks. It was embedded in paraffin and cut into 6 ${\mu}m$ thickness. The sections were stained with H & E and observed by optical microscope. The results were as follows; 1. The periosteum played an important role in osteointegration of $Bio-Oss^{(R)}$ in bone defects. 2. When the periosteum remained intact and $Bio-Oss^{(R)}$ was placed on the defect, $Bio-Oss^{(R)}$ with periosteum covering has been incorporated into the newly formed bone from 2-week postoperatively. 3. When the periosteum was removed at the surgical procedure, invasion of connective tissue took place among the granules, and new bone formation was delayed compared to periosteum covering group. Therefore, when the bone grafting was performed with periosteal incision procedure to achieve tension-free suture, the integrity of the overlying periosteum should be maintained to avoid fibrous tissue ingrowth.

• Journal of Life Science
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• v.17 no.10
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• pp.1347-1353
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• 2007

The changes of glycoconjuagates (GCs) in rat kidney due to maturation were studied from samples of fetal and postnatal kidneys by lectin histochemistry. Rat kidneys of perinatal ages and adults were fixed in 4% paraformaldehyde and were stained with nine kinds of biotinylated lectins. The immature forms of the renal developmental stage such as vesicles and ureteric bud were observed in the cortex as late as day 14 of postnatal life, but the histological appearance of the weaning kidney was similar to that observed in adults. As for histochemical properties of GCs in the glomeruli, Con A affinity tended to increase with aging, but both RCA-1 and LCA affinities showed a transient increase in immature glomeruli of neonatal rats. DBA affinity with SBA, PNA, BSL-1 and RCA-1, additional Con A one in proximal tubule, were increased in both proximal and distal tubules according to maturation. In contrast to this, transient intensive LCA affinity were demonstrated in immature proximal and distal tubule of neonatal rats. In the collecting tubules, DBA, SBA, PNA and sWGA affinities tended to increase according to maturation, but transient increase for BSL-1, RCA-1 and LCA affinities were detected in neonatal rats. The present results suggest that the mature glycosylation pattern of the kidney undergoes profound changes during maturation and is probably associated with functional maturation of the kidney.