• Journal of Life Science
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• v.19 no.12
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• pp.1685-1689
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• 2009

Phytochelatins (PCs) are small polypeptides synthesized by PC synthase (PCS). They are present in various living organisms including plants, fission yeast, and some animals. The presumed function of PCs is the sequestration of cytosolic toxic heavy metals like cadmium (Cd) into the vacuoles via vacuolar membrane localized heavy metal tolerance factor 1 (HMT-1). HMT-1 was first identified in fission yeast (SpHMT-1), and later in Caenorhabdtis (CeHMT-1). Recently, its homolog has also been found in PC-deficient Drosophila (DmHMT-1), and this homolog has been shown to be involved in Cd detoxification, as confirmed by the heterologous expression of DmHMT-1 in fission yeast. Therefore, the dependence of HMT-1 on PC in Cd detoxification should be re-evaluated. I heterologously expressed SpHMT-1 in cytosolic PC-deficient yeast, Saccharomycea cerevisiae, to understand the dependence of HMT-1 on PC. Yeast cells expressing SpHMT-1 showed increased tolerance to Cd compared with control cells. This result indicates that SpHMT-1 is not strictly correlated with PC production on its function. Moreover, yeast cells expressing SpHMT-1 showed increased tolerance to exogenously applied glutathione (GSH) compared with control cells, and the tolerance to Cd was further increased by exogenously applied GSH, while tolerance in control cells was not. These results indicate that the function of SpHMT-1 in Cd detoxification does not depend on PCs only, and suggest that SpHMT-1 may sequester cytosolic GSH-Cd complexes into the vacuole.

• Journal of Food Hygiene and Safety
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• v.28 no.2
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• pp.188-193
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• 2013

Anatoxin-a, saxitoxin and neosaxitoxin are produced by Aphanizomenon flos-aquae that is a sort of the cyanobacteria phylum. Therefore, it is not permitted for food materials in Korea. Traditionally, the classification of cyanobacteria has been based on morphological characters such as trichome width, cell size, division planes, shape, and the presence of character such as gas vacuole. But, some diagnostic features, such as gas vacuole or akinetes, can show variation with different environmental or growth conditions and even be lost during cultivation. Therefore, we developed detection method for functional foods containing Aph. flos-aquae by PCR. To design the primer, 16S rRNA region of Aph. flos-aquae, Spirulina laxissima, and Spirulina spp. registered in the GeneBank (www.ncbi.nlm.nih.gov) have been used and for comparative analysis, BioEdit ver. 7.0.9.0. was used. As a result, we was design AFA-F1/AFA-R1 (363 bp) primer for the differentiation Aph. flos-aquae from chlorella, spirulina, green tea, and spinach. Also, it could be distinguished chlorella and spirulina products those are made to contain 1% Aph. flos-aquae.

• Korean Journal of Environmental Biology
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• v.32 no.3
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• pp.225-233
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• 2014

In this study, we investigated the changes in the physiological and histological traits of a sea squirt (Halocynthia roretzi) with the emergence of the soft tunic syndrome induced by the water temperature control (6, 9, 12, 15, 18, 21, 24 and $27^{\circ}C$). It was observed that the induction rate of the soft tunic syndrome was highest at $15^{\circ}C$, but lowest at $24^{\circ}C$. Based on the tunic color condition and contraction strength, the whole process were classified into 4 stages as S0, S1, S2 and S3. Interestingly, there were significant differences in oxygen consumption and filtration rate were observed during S0-S3. The most distinctive aspects were change of blood cell composition at stage S3, whereas multi-vacuole cell ratio was decreased by 1/2 and morula cell ratio expanded about 10 times during S0-S3. Further, change of organ structure started following the syndrome such as degeneration of epithelial cells, microfilaments, increment in hemocytes and damage in muscle fiber have been detected in tunic, siphon, branchial sac, body wall musculature and pyloric gland. Briefly, our study results indicated that the normal physiological functions of the sea squirt can be affected due to the soft tunic syndrome induced by water temperature.

• Applied Microscopy
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• v.28 no.4
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• pp.603-613
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• 1998

The present study was designed in order to observe relationship between the endoplasmic reticulum and the Golgi complex during spermiogenesis of the long-fingered bat (Miniopterus schreibersi fuliginosus). The testes were obtained from adult bats and treated with the prolonged osmification or fixed with ferrocyanide reduced osmiun. In the Golgi phase, The Golgi complex shows an oval shape, and was composed of a cortex and a medullar enclosing acrosome. The Golgi vacuoles with electron-dense granules of crescent shape were fused with each other. The smooth endoplasrnic reticulum was scattered in all the area of the cytoplasm. In the cap phase, The Golgi complex was crescent in shape, and faced to a nucleus. Large and small vesicles were fused with each other, and then fused with a acrosomal vacuole. The rough endoplasmic reticulum was close to the large Golgi vacuole. In the acrosome phase, The Golgi complex was moved to behind of the acrosome face. Small vesicles were fused with an acrosome, and cisternae of the trans-face of Golgi complex was connected with an acrosome in the early acrosome phase. The smooth endoplasmic reticulum was distributed in the cytoplasm. The annulate lamellar was originated from a radial body-annulate lammellae complex. In the maturation phase, The Golgi complex with dilated cistrern appeared in the cytoplasm, and also, annulate lamellar was observed in the cytoplasm. The connection of the annulate lamellar with the cistern of radial body suggests that an annulate lamellar seems to be closely related to radial body. The smooth endoplasmic reticulum was scattered in the cytoplasm in the early Golgi phase, but annulate lamellar-radial body complex which might be a residual and disappearing form of the smooth endoplasmic reticulum appeared in the acrosome phase. The Golgi complex steadily remained in the late maturation phase when the endoplasmic reticulum began to disappear from the cytoplasm: the Golgi complex was still occurred after acrosome formation. The observations obtained in the present study, which was characterized by the presence of the Golgi complex in the late maturation phase, suggests that the Golgi complex may play an important role also even after the acrosome formation.

• Korean Journal of Oriental Medicine
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• v.1 no.1
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• pp.477-493
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• 1995

In order to facilitate the molecular characterization of the Herpes simplex Virus types 1 and types 2 genome DNAs, a gene library of cloned restriction frtgments have been produced. The Vero cells were infected with HSV-1 and HSV-2. 48 hours after infection, the infected cells Ivere Iysed, and multinucleated giant cells were observed approximately at seventy-two hours postinfection. The multiplication of HSV-1 and HSV-2 was observed in Vero cells using electromicroscopy. The nucleocapsids in nuclei were obseryed, and the assembled virions were budded out through the vacuole, and the virions were released from the cells. HSV-1 and HSV-2 was analyzed by digestion of their genome DANs with restriction ensymes. HSV-1 and HSV-2 genome DNAs were digested with BarnHI, Bgfl respectively. The BarnHI rlestriction fragments of HSV-1 and HSV-2 genome DNAs were twenty-seven fragments and thair molecular sizes were ranging $0.70{\sim}15.08$, $4.4{\sim}31.0$ tilobases. The BglII restriction fragments of HSV-1 and HSV-2 genome DNAs were sixteen, eighteen fragments and thair molecular sizes were ranging $4.8{\sim}30.0$, $1.2{\sim}25.0$ kilobases. And then BglII restriction frgments were cloned in Escherichia coli(E.coil) using the plasmid vector pBacPAK9.

• Applied Microscopy
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• v.12 no.1
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• pp.49-56
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• 1982

A case of systemic cryptococcosis developed in 4 year old boy was described and illustrated by light and electron microscope. Light microscopically, the upper dermis of the skin showed chronic nonspecific inflammation with numerous spherical spores surrounded by a clear halo created by the wide gelatinous capsule. Ultrastructurally, the C. neoformans showed the wide capsule containing microfibrils that appeared to radiate from the cell wall and to coil and interwine in various directions. The cell was uninucleate with a single nucleolus. Along the inner nuclear envelope, numerous small vesicles were present. In addition, C. neoformans presented membranous organelles derived from the plasma membrane and comparable to bacterial mesosomes.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.2
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• pp.189-195
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• 2017

This study aimed to compare the cellular toxicities of three clinically used dry eye treatments; 3% diquafosol tetrasodium and hyaluronic acid at 0.3 and 0.18%. A methyl thiazolyltetrazoiun (MTT)-based calorimetric assay was used to assess cellular proliferation and a lactate dehydrogenase (LDH) leakage assay to assess cytotoxicity, using Human corneal epithelial cells (HCECs) exposed to 3% diquafosol tetrasodium, 0.3% hyaluronic acid (HA), or 0.18% HA or 1, 6 or 24 h. Cellular morphology was evaluated by inverted phase-contrast light microscopy and electron microscopy, and wound widths were measured 24 h after confluent HCECs were scratched. Diquafosol had a significant, time-dependent, inhibitory effect on HCEC proliferation and cytotoxicity. HCECs treated with diquafosol detached more from the bottoms of dishes and damaged cells showed degenerative changes, such as, reduced numbers of microvilli, vacuole formation, and chromatin of the nuclear remnant condensed along the nuclear periphery. All significantly stimulated reepithelialization of HCECs scratched, which were less observed in diquafosol. Therefore, epithelial toxicity should be considered after long-term usage of diquafosol and in overdose cases, especially in dry eye patients with pre-existing punctated epithelial erosion.

• Biomedical Science Letters
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• v.23 no.1
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• pp.44-47
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• 2017

S100A8 and S100A9 are associated with myeloid cell differentiation, chemotactic activities, adhesion of neutrophils, and apoptosis. In this study, we investigated the contribution of S100A8 and S100A9 to differentiation of the human eosinophilic leukemia cell line, EoL-1. S100A8 and S100A9 increased the number of vacuole per one cell and the protein expression of EPO and MBP. Rottlerin, an inhibitor of protein kinase C delta ($PKC{\delta}$), inhibited the EoL-1 cell differentiation induced by S100A8 and S100A9. These results suggest that S100A8 and S100A9 may regulate the differentiation of eosinophilic progenitors. Moreover, these findings may shed light on elucidation of eosinophil differentiation due to S100 proteins.

• Mycobiology
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• v.46 no.2
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• pp.114-121
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• 2018

Mon1 is a guanine nucleotide exchange factor subunit that activates the Ypt7 Rab GTPase and is essential for vacuole trafficking and autophagy in eukaryotic organisms. Here, we identified and characterized the function of Mon1, an ortholog of Saccharomyces cerevisiae Mon1, in a human fungal pathogen, Cryptococcus neoformans. Mutation in mon1 resulted in hypersensitivity to thermal stress. The mon1 deletion mutant exhibited increased sensitivity to cell wall and endoplasmic reticulum stress. However, the mon1 deletion mutant showed more resistance to the antifungal agent fluconazole. In vivo studies demonstrated that compared to the wild-type strain, the mon1 deletion mutant attenuated virulence in the Galleria mellonella insect model. Moreover, the mon1 deletion mutant was avirulent in the murine inhalation model. These results demonstrate that Mon1 plays a crucial role in stress survival and pathogenicity in C. neoformans.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.75.2-75
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• 2003

To develop an effective protection strategy against tomato bushy stunt virus (TBSV), tobacco plants expressing single-chain Fv antibodies (scFv), were established. A previous had shown that the replication activity of viral replicase was inhibited by the selected scFvs. Moreover, no systemic symptom was found after virus inoculation on leaves of wt N. benthamiana infiltrated with an Agrobacterium suspension resulting i3l expression of the scFvs. However, control plants showed systemic symptoms. In this study the localization of the scFvs within two transgenic plant lines, (CP28H3, CP-P55) was demonstrated using immunogold labelling. The gold particles, indicating the presence of scFv, were mostly found In the cytoplasm of the plant cells including chloroplasts and in the cell walls. However, they were hardly found in the vacuole, nucleoplasm and intercellular spaces. Gold particles often accumulated in either the cytosol or chloroplasts showing a specific labeling, There was no difference in type of gold labeling between both transgenic lines. The localization of the scFv in the cytoplasm further conforms the inhibition of the RNA-dependent RNA polymerase (RdRp) by the selected scFv because it is known that the RdRp is localized to membraneous cytosolic structures.