• KSBB Journal
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• v.19 no.2
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• pp.118-124
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• 2004

Melanocytes synthesize melanin within discrete organelle termed melanosomes which are transferred to the surrounding keratinocytes and can be produced in varying sizes, numbers and densities. Skin whitening products have become increasingly popular in the past few years. The most successful natural skin whitening agents are: Arbutin, Vitamin C, Kojic acid, Mulberry, which are all tyrosinase inhibitors. In this work, melanoston, a melanogenesis inhibitor isolated from yeast was studied to understand its mechanism of melanogenesis inhibition. It was found that melanoston was not a tyrosinase inhibitor, while when melanoston was applied to the B16 melanoma cell culture media, the intracellular tyrosinase activity was decreased by more than 30％, When B16 melanoma was stimulated with ${\alpha}$-MSH, cell morphololgy was dramatically changed to have lots of dendrites on the cell membrane surface. On the other hand, B16 was treated with ${\alpha}$-MSH and melanoston, simultaneously, the change of cell morphology was not so great. This inhibition effect of melanoston was found to be related to the inhibition of intracellular activation and transportation of tyrosinase, which was observed by immunostaining of B16 melanoma using anti-tyrosinase antibody. From these results, melanoston was regarded as an inhibitor to the differentiation of melanoma cells.

• Journal of Yeungnam Medical Science
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• v.2 no.1
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• pp.175-181
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• 1985

Agrobacterium tumefaciens induces cancerous growths called crown galls at wound sites on dicotyledonous plants. A large plasmid called Ti plasmid is responsible for virulence. Upon tumor induction, part of the plasmid, termed T-DNA, becomes integrated into plant genome and its genetic sequences are expressed. These properties allow Ti plasmids to be used as gene vectors in plants. Several in vitro methods for the transfer of Ti plasmid into plant cell have been developed. One of them is the treatment of bacterial spheroplasts and plant protoplasts mixture with polyethylene glycol that is generally used as fusogen in cell-to-cell fusion. Several workers investigated the interaction of bacterial spheroplasts with plant protoplasts in the presence of polyethylene glycol and suggested that the interaction is not fusion but endocytosis. In this report we observed the interaction of Agrobacterium tumefaciens spheroplasts with Nicotiana tabacum protoplasts by electron microscope. Agrobacterium tumefaciens spheroplasts and Nicotiana tabacum protoplasts were prepared and mixed in the presence of polyethylene glycol and high pH-high $Ca^{2+}$ buffer. Then the interaction of the spheroplasts with the protoplasts was examined by transmission electron microscope. After the treatment of polyethylene glycol the spheroplasts adhered to the surface of the protoplasts and then they were engulfed by the protoplasts. After the high pH-high $Ca^{2+}$ buffer treatment the engulfed spheroplasts lost their cell integrity. No fusion process was observed. Thus all these observations suggest that the introduction process of Agrobacterium tumefaciens spheroplasts into Nicotiana tabacum protoplasts with the aid of polyethylene glycol is endocytosis.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.195-198
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• 2006

Lethal toxin is a critical virulence factor of anthrax. It is composed two protein: protective antigen (PA) and lethal factor (LF). PA binds to specific cell surface receptors and, forms a membrane channel that mediates entry of LF into the cell. LF is a zinc-dependent metalloprotease, which cleaves MKKs [MAPK (mitogen-activated protein kinase) kinases] at peptide bonds very close to their N-termini. In this study, we suggest application of cell-based assays in the early phase of drug discovery, with a particular focus on the use of yeast cells. We constructed MEK1 expression system in yeast to determine LF activity and approached cell-based assay system to screen inhibitors, in which the results covering the construction of LF-substrate in yeast expression vector, expression, and LF-mediated proteolysis of substrate were described. These results could provided the basic steps in design of cell-based assay system with the high efficiency, rapidly and easy way to screening of inhibitors.

• Applied Microscopy
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• v.15 no.2
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• pp.111-129
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• 1985

The morphology and histology of the gastrointestinal mucosa of the mole, Talpa micrura coreana (Thomas), were studied using light and scanning electron microscopes. Tissue specimens were taken from body and pyloric portions of the stomach, and from the initial, proximal, middle, distal and terminal portions of the intestine. For light microscopy, tissue blocks were fixed in 10% buffered neutral formalin, embedded in paraffin wax, and sectioned at a thickness of $5{\mu}m$. These sections were stained with hematoxylin-eosin. For scanning electron microscopy, tissue blocks were fixed in 1% glutaraldehyde-1.5% paraformaldehyde, and postfixed in 1% osmium tetroxide, dehydrated in graded alcohol, transferred to isoamylacetate and dried by the critical point drier(Polaron E 3000). Subsequently, specimens were coated with gold and observed with a JSM-35C scanning electron microscope. The results were as follows: The mucous membrane of the body portion of the stomach had numerous irregular folds and the pyloric mucosa formed the strawberry-shaped folds, and general histological structures of each portion were similar to those of man. The intestine could not be differentiated macroscopically and microscopically into small and large intestines. There was no cecum, appendix, taenia coli, haustra coli or appendices epiploicae. In the initial portion (4 mm long), conical or tongue-shaped villi with the height of $143.3{\pm}10.7{\mu}m$ were present, and large mucous glands were seen in the submucosa. In the proximal, middle and distal portions, wavy folds composed of the epithelium and lamina propria were densely and transversely arranged, and their heights were $440.4{\pm}45.5{\mu}m,\;454.4{\pm}19.9{\mu}m\;and\;205.2{\pm}33.5{\mu}m$, respectively. The mucosa of the terminal portion (3 cm long) formed several longitudinal folds, and the intestinal glands were directly opened on the smooth surface of the folds. Aggregated lymphoid follicles were observed in the major portions of the intestine except the initial and terminal portions. There was no circular or semilunar fold throughout the intestine.

• Applied Microscopy
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• v.29 no.2
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• pp.177-187
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• 1999

Ultrastructure of gametes in the three-spine stickleback, Gasterosteus aculeatus aculeatus was observed, utilizing light, scanning and transmission electron microscopes. The egg of three-spine stickleback is spherical and demersal type. The eggs are highly adhesived to each other but not to substrates. There are many oil droplets in vitelline membrane. The outer surface of egg envelope is arranged by mushroom-like structures and pore canals. The egg have a micropyle, sperm entry site, in the area of the animal pole. The egg envelope consists of three layers, an outer layer with high electron density, a middle layer consisting two layers and an inner layer consisting of 16 to 20 layers. In the fertilized egg envelope, the molecular weights of these components ranged from 14 kDa to 205 kDa. The molecular weights of nam protein bands are 19.4 kDa, 36.7 KDa, 39.4 kDa, 42.9 kDa, 46.1 kDa and 53.0 kDa. The head of spermatozoa is spherical shape and the acrosome is absent. The mitochondria in midpiece are arranged from one to three layers and separated from the axoneme by the cytoplasmic canal. The tail has two lateral fins and the axoneme is of the 9+2 structure.

• Applied Microscopy
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• v.37 no.2
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• pp.65-72
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• 2007

The oogenesis and ultrastructure of fertilized egg envelope of false dace were investigated by light and electron microscope. The cytoplasm of false dace oogonia was basophilic and many nucleoli were located at inner side of nuclear membrane. In primary oocytes, yolk vesicles were distributed in marginal area only and egg envelope was not formed on egg outside. In secondary oocyte, the egg envelope was formed and yolk vesicles were increased than that of early stage in cytoplasm. The amount of basophilic substance was decreased. In case of matured egg, thickness of egg envelope and site of egg were increased, basophilic substance was distributed in egg envelope around only. The yolk vesicles were changed to yolk mass in accordance with development. The fertilized egg was of ellipsoidal, adhesive type and yellowish, have a single micropyle in the area of the animal pole. The fertilized egg envelope consisted of three layers, an outer adhesive layer, a middle layer consisting of 6 lamellae alternating layers and an inner electron dense layer. An outer surface of the fertilized egg envelope was arranged by adhesive fibrous structures. In conclusion, it is summarized that the oogenesis of false dace were the increase of cell size, the formation and accumulation of yolk, and decrease of basophilic intensity in cytoplasm. These ultrastructural characteristics of fertilized egg envelope from false dace can be utilized in taxonomy of teleost.

• Applied Microscopy
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• v.39 no.3
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• pp.237-243
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• 2009

Chinese minnow, Rhynchocypris oxycephalus is a teleost belonging to Leuciscinae, Cyprinidae. The oogenesis and ultrastructure of egg envelope in Chinese minnow were investigated by light and electron microscopes. The ovary was of white yellowish and ellipsoidal shape with the major axis 30 mm and the minor axis 7mm. Cytoplasm of oogonia was basophilic and many nucleoli were located at inside of nuclear membrane. In primary oocytes, yolk vesicles were distributed only in the marginal area and egg envelope was not formed on the outside of an egg. In secondary oocytes, the egg envelope was formed and yolk vesicles in the cytoplasm were increased than the earlier stage. The basophilic substance of cytoplasm was changed to acidic. In case of matured egg, thickness of egg envelope and size of egg were increased. The yolk vesicles were changed to yolk mass in accordance with development. The outer surface of egg envelope was covered by microvilli-structures, and had a micropyle on the area of animal pole. Egg envelope consisted with 2 layers, an adhesive outer layer with microvilli-structures and fibrillar inner layer. In conclusion, the oogenesis of Chinese minnow was characterized by the increase in cell size, the formation and accumulation of yolk, and the decrease of basophilic substance in the cytoplasm. The oogenesis of Chinese minnow seems to share common patterns in Cyprinidae, but these ultrastructural unique characters of egg envelope can be utilized in taxonomy of teleost.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1343-1349
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• 2012

Most of the Helicobacter pylori strains containing the cag pathogenicity island (PAI) have been associated with more severe gastric disease in infected humans. The cag PAI is composed of 27 proteins, and some of the components are required for CagA translocation into host cells as well as induction of proinflammatory cytokines, such as interleukin-8 (IL-8); however, the exact function of most of the components remains unknown or poorly characterized. In this study, we demonstrated that CagT (HP0532), which is an essential structural component of the cag PAI apparatus, plays an important role in the translocation of CagA into host epithelial cells. In addition to being located on the bacterial surface, CagT is also partially localized in the inner membrane, where it acts as a chaperone-like protein and promotes CagA translocation. However, CagT secretion was not detected by immunoprecipitation analysis of cell culture supernatants. Meanwhile, CagT was related to the introduction of IL-8 of the host cell. These results suggest that CagT is expressed on both the inner and outer bacterial membranes, where it serves as a unique type IV secretion system component that is involved in CagA secretion and cag PAI apparatus assembly.

• Mycobiology
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• v.45 no.4
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• pp.297-311
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• 2017

Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin $[IL]-1{\beta}$, tumor necrosis factor ${\alpha}$, IL-6, IL-8, interferon ${\gamma}$, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules ($CD8^+$ and $CD4^+$) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as $200{\mu}g/mL$ and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.

• Archives of Plastic Surgery
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• v.37 no.4
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• pp.469-472
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• 2010

Purpose: Aplasia Cutis Congenita (ACC) is a rare disease characterized by the focal defect of the skin at birth, frequently involving scalp, but it may affect any region of the body. There are no etiology known but some conditions such as intrauterine vascular ischemia, amniotic adherences and viral infections are associated. The ideal treatment for the ACC is not known. Superficial and relatively small sized defects (< $3{\times}5\;cm$) may heal spontaneously and large defects related with risks of infection and bleeding may require aggressive surgical treatment. Hyalomatrix$^{(R)}$ is a bilayer of an esterified hyaluronan scaffold beneath a silicone membrane. It has been used as a temporary dermal substitute to cover deep thickness skin defect and has physiological functions derive from the structural role in extracellular matrix and interaction with cell surface receptor. This material has been used for the wound bed pre-treatment for skin graft to follow and especially in uncooperative patient, like a newborn, this could be a efficient and aseptic way of promoting granulation without daily irritative wound care. For this reason, using Hyalomatrix$^{(R)}$ for the treatment of ACC was preferred in this paper. Methods: We report a case of a newborn with ACC of the vertex scalp and non-ossified partial skull defect. The large sized skin and skull defect ($6{\times}6\;cm$) was found with intact dura mater. No other complications such as bleeding or abnormal neurologic sign were accompanied. Escharectomy was performed and Hyalomatrix$^{(R)}$ was applied for the protection and the induction of acute wound healing for 3 months before the split-thickness skin graft. During the 3 months period, the dressing was renewed in aseptic technique for every 3 weeks. The skin graft was achieved on the healthy granulation bed. Results: The operative procedure was uneventful without necessity of blood transfusion. Postoperative physical examination revealed no additional abnormalities. Regular wound management was performed in out-patient clinic and the grafted skin was taken completely. No other problems developed during follow-up. Conclusion: Hyalomatrix$^{(R)}$ provides protective and favorable environment for wound healing. The combination of the use of Hyalomatrix$^{(R)}$ and the skin graft will be a good alternative for the ACC patients with relatively large defect on vertex.