• Journal of Life Science
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• v.32 no.12
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• pp.956-961
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• 2022

Plant Chloroplast have several advantages as an expression platform of biopharmaceuticals over conventional expression platforms such as mammalian cells, yeast and bacteria. First, plants do not serve as a host for mammalian infectious virus and have endotoxin like bacteria which can cause anaphylactic shock. In addition, high copy number of chloroplast genome allows for chloroplast transformants to reach the high level of expression of heterologous genes. Moreover, the integration of transgenes into specific region of chloroplast genomes makes chloroplast transformants unaffected by positional effect which can be frequently observed from nuclear transformants, resulting in loss of transgene expressions. Antimicrobial peptides (AMPs) are a kind of innate immunity which is found from bacteria to humans. Unlike conventional antibiotics, very less dosage of AMPs can have catastrophic effect on bacterial survival. Further, the repeated use of AMPs does not trigger the development of bacterial resistance. Moricin, one of the AMPs, was isolated from Bombyx mori, a silkworm moth. The C-terminal of moricin consists largely of basic amino acids, and the N-terminal has an α-helix structure. Moricin was chosen and expressed in a SUMO/SUMOase without leaving any unwanted amino acids which could potentially affect the anti-bacterial activity of the moricin. The transformation vector used in this study has already been created in this lab for the expression in both prokaryotic systems such as E. coli and chloroplast. The expressed moricin was purified using Ni columns and SUMOase, and the antibacterial activity of the purified moricin was confirmed using an agar diffusion assay.

• Journal of Life Science
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• v.27 no.1
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• pp.15-22
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• 2017

Streptococcus mutans (S. mutans), which plays a major role in the etiology of human dental caries, is able to tolerate exposure to acid shock in addition to its acidogenicity. We investigated the effects of pH stress on membrane fluidity, activities and expression levels of F-ATPase, and proton permeability in S. mutans. Using 1,6-diphenyl-1,3,5-hexatriene, we observed membrane ordering at pH 4.8 and pH 8.8. The ordering effects were larger at pH 4.8 in cytoplasmic membranes isolated from S. mutans (CMSM). Increasing pH resulted in a decrease in the activities and expression levels of F-ATPase. The proton permeability was decreased at both acidic and alkaline pHs, and the lowest permeability was observed at pH 4.8. The lower permeability at pH 8.8 than pH 6.8 is likely to be caused by the decreased proton influx due to the decreased CMSM fluidity. In addition, it seems to be evident that extremely low permeability at pH 4.8 was caused by the decreased proton influx due to the decreased CMSM fluidity as well as the increased proton efflux due to the increased activity and expression level of F-ATPase. It is likely that CMSM fluidity and F-ATPase activity are two major key factors that determine proton permeability in S. mutans. We suggest that CMSM fluidity plays an important role in the determination of proton permeability, which sheds light on the possibility of using nonspecific membrane fluidizers, e.g., ethanol, for anti-caries purposes.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.33-45
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• 2011

Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of cancer cell. Inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the cancer cell was reported. However, its role during oocyte maturation and early embryo development is very insufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on meiotic maturation and early embryonic development in pigs. We also investigated several indicators of developmental potential, including structural integrity, gene expression (Hsp90-, cell cycle-, and apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Then, we examined the roles of Hsp90 inhibitor on viability of primary cells in pigs. Porcine oocytes were cultured in the NCSU-23 medium with or without 17-AAG for 44 h. The proportion of GV arrested oocytes was significantly different between the 17-AAG treated and untreated group (78.2 vs 34.8%, p<0.05). After completion of meiotic maturation, the proportion of MII oocytes was lower in the 17-AAG treated group than in the control group (27.9 vs 71.0%, p<0.05). After IVF, the percentage of penetrated oocytes was significantly lower in the 17-AAG treated group (25.2%), resulting in lower normal pronucleus formation (2PN of 14.6%). Therefore, the inhibition of meiotic progression by Hsp90 inhibitor played a critical role in fertilization status. Porcine embryo were cultured in the PZM-3 medium with or without 17-AAG for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (7.5 vs 4.4, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. The mRNA expressions of cell cycle-related genes were down-regulated in the 17-AAG treated group compared with control. Also, the expression of the pro-apoptotic gene Bax increased in 17-AAG treated group, whereas expression of the anti-apoptotic gene Bel-XL decreased. However, the expression of ER stress-related genes did not changed by 17-AAG. Cultured pESF cells were treated with or without 17-AAG and used for MIT assay. The results showed that viability of pESF cells were decreased by treatment of 17-AAG ($2{\mu}M$) for 24 hr. These results indicated that 17-AAG decreased cell proliferation and increased cell death. Expression patterns Hsp90 complex genes (Hsp70 and p23), cell cycle-related genes (cdc2 and cdc25c) and apoptosis-related genes (Bax and Bcl-XL) were significantly changed by using RT-PCR analysis. The spliced form of pXbp-1 product (pXbp-1s) was detected in the tunicamycin (TM) treated cells, but it is not detected in 17-AAG treated cells. In conclusion, Hsp90 appears to play a direct role in porcine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with cell cycle- and apoptosis-related genes expression in developing porcine embryos.

• Tuberculosis and Respiratory Diseases
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• v.41 no.5
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• pp.484-488
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• 1994

Background : The antigen-specific receptor on the surface of most peripheral T lymphocytes is a disulfide-linked heterodimer composed of $\alpha$ and $\gamma$ subunits, noncovalently associated with CD3 polypeptides. Recently, a novel type of CD3-associated heterodimer was described on a T cell subset that does not express CD4 or CD8 molecules. This second type of TCR dimer is composed of chains encoded for by the $\gamma$- and $\delta$-TCR genes. These cells may exert both cytotoxic and lymphokine producing functions. Although it was reported that some ${\gamma}{\delta}$-TCR might recognize an MHC-linked determinant, the funεtion or physiologic ligand for this new receptor is not yet clear. It was found that ${\gamma}{\delta}$-TCR can react with 65 kD heat shock protein of M. tuberculosis, which suggests the possible protective role of ${\gamma}{\delta}$ T lymphocytes against tuberculosis. In our previous study, there was neither the increase in number nor the functional activation of ${\gamma}{\delta}$ T cells in the peripheral blood from patients with pulmonary tuberculosis. Now we report the distribution of ${\gamma}{\delta}$ T cells in the regional sites of M. tuberculosis infection, especial1y tuberculous lymphadenitis. Methods : Lymph nodes from patients with pathologically-proven tuberculous lymphadenopathy (n=5) and reactive hyperplasia (n=3) were used. Tissues were frozen in liquid nitrogen immediately after removal and stored below $-70^{\circ}C$. The cryostat sections of these frozen specimens were stained with anti-Leu-4 Ab, Identi-T TCR ${\delta}1$, and Identi-T ${\beta}F1$. The number of positively stained cells were counted at high power field. Results : The infiltration of ${\gamma}{\delta}$ T cells was significantly higher in the lymph nodes from patients with tuberculous lymphadenopathy than that with reactive hyperplasia ($16.3{\pm}10.3%$ vs. $1.7{\pm}1.5%$). Conclusion : These results suggest that ${\gamma}{\delta}$) T cells may play a role in the defense against M. tuberculosis infection, especially in the regional sites of infection.

• Proceedings of the Ginseng society Conference
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• 1988.08a
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• pp.92-98
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• 1988

The present study was performed to find the effects of ginseng and its saponins. which is written in Chung Yao Ta Tsu Tien as anti-amnesia in its chief indication. on experimental amnesia in mice. In the step through test. ginsenoside $Rb_1\;(GRb_1)\;and\;GRg_1$ facilitated the registration of memory and antagonized the electroconvulsive shock (ECS)-induced inhibition of the retention of memory. Moreover. $GRg_1$ antagonized the EtOH-induced inhibition of the retrieval of memory. In the step down test. $GRb_1\;GRb_2\;and\;GRg_1$ antagonized the ECS-induced inhibition of the retention of memory. Moreover. $GRg_1$ antagonized the EtOH-induced inhibition of the retrieval of memory and facilitated the acquisition of short term memory. In the shuttle hox and lever press tests. they have no effects on acquisition and retrieval of memory. except $GRb_1\;GRb_1$ depressed the retrieval of conditioned avoidance response in the shuttle box test. After the end of four tests. the effects of these orally administered drugs on sedative. analgesic. antipyretic and anticonvulsant actions. and on spontaneous and exploratory movements were tested in doses of less than 500mg/kg. but they had none of these effects. Present study may indicate that $GRg_1$ had effects on the retrieval of memory and on the acquisition process of learning response. The recent research on the role of NGF for the survival. regeneration and regulation of brain in adult animals. indicated the importance of NGF on dementia and amnesia. During our research on the specificity of the neurite out growth induced by NGF. we found that the effect of NGF was potentiated by $GRb_1$ in organ cultures of chick embryonic dorsal root ganglia. Then. the effect of $GRb_1$ on neuronal cell survivalin cell culture system was studied. $GRb_1$ potentiated the NGF-mediated increase of neurofilaments in cell cultures of chick embryonic sensory and sympathetic neurons. NGF with $GRb_1$ also showed a tendency to increase the number of surviving neurons of rat embryonic cerebral cortex. NGF increased choline acetyl transferase activity in cell cultures of rat embryonic septum area neurons. but $GRb_1$ did not potentiate NGF activity in cell cultures of rat embryonic septum area neurons. Present study may indicate that $GRb_1$ plays an important role for the survival or regeneration of neurons in the brain.

• Journal of the Korean Institute of Landscape Architecture
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• v.40 no.2
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• pp.1-14
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• 2012

The purpose of this study is to derive a desirable riverfront construction plan to me for the activities of citizens through the evaluation and analysis of the urban riverfront space from the perspective of universal design. Therefore, previous studies were examined in order to induce evaluation tools that bhve been grafted from the universal design concept; in addition, a field survey was conducted in Shincheon, Daegu, which was selected as the study target, in order to induce problems and improvement directions from the perspective of universal design. The major results can be summarized as follows. (1) In the 'fairness' aspect, all the items such as installation of integrated functional signage, showing pictures, symbols, foreign language signs, and restroom entrances signage were determined to be 'All Unsuitable' for all sections; and therefore, it is necessary to improve the fairness of usage for everybody. (2) In the 'Functionality(Usability)' aspect, all items such as installation of bicycle paths beside access roads, installation of integrated functional signs, and night light signs were determined to be 'All Unsuitable' for all sections; therefore, it is necessary to improve the functionalities of these facilities. (3) In the 'Convenience' aspect, all items such as the installation of bicycle parking areas, continuous rest facilities, and back and ann support(handles) at resting facilities were determined to be 'All Unsuitable' for many sections; therefore, it is necessary to improve these facilities for the convenience of usage. (4) In the 'Information(Recognizability)' aspect, all items such as showing pictures, symbols, foreign languages and installation of night light signs, and restroom entrances signage were determined to be 'All Unsuitable' for all sections; therefore, it is necessary to improve the recognizability to minimize misunderstandings and confusion. (5) In the 'Safety' aspect, all items such as the installation of safe pedestrian paths in parking areas, using anti-slip and shock absorption materials on restroom floors, and the continuous installation of pedestrian paths that are separate from bicycle paths were determined to be 'All Unsuitable' for all sections; therefore, it is necessary to improve the safety to prevent accidents. (6) In the 'Amenity' aspect, access roads, parking areas, hygiene facilities, convenience facilities, and waterside facilities for many sections were determined to be 'All Unsuitable'; therefore, it is necessary to conduct more concentrated hygiene management. (7) In the 'Accessibility(Mobility)' aspect, all items such as the installation of safe pedestrian paths in parking areas, and continuous pedestrian paths that are separate from bicycle paths were determined to be 'All Unsuitable' for all sections; therefore, it is necessary to improve the accessibility to provide safety and convenience. (8) In the 'Durability' aspect, access roads, parking areas, rest facilities, convenience facilities, fitness facilities, and waterside facilities were determined to be 'All Unsuitable' for many sections, therefore, it is necessary to improve sunken or damaged areas by inspecting facilities by section.