• Analytical Science and Technology
• /
• v.31 no.5
• /
• pp.208-218
• /
• 2018

Autophagy is a cellular process whereby cytosolic materials or organelles are taken up in a double-membrane vesicle structure known as an autophagosome and transported into a lysosome for degradation. Although autophagy has been studied at the genetic, cellular, or biochemical level, systematic ultrastructural quantitative analysis of autophagosomes during the autophagy process by using transmission electron microscopy (TEM) has not yet been reported. In this study, we performed ultrastructural analysis of autophagosomes in wild-type (WT) mouse embryonic fibroblasts (MEFs) and autophagy essential gene (atg5) knockout (KO) MEFs. First, we performed ultrastructural analysis of autophagosomes in WT MEFs compared to atg5 KO MEFs in basal autophagy or starvation-induced autophagy. Although we observed phagopore, early, late autophagosomes, or autolysosomes in WT MEFs, atg5 KO MEFs had immature autophagosomes that showed incomplete closure. Upon starvation, late autophagosomes accumulated in WT MEFs while the number of immature autophagosomes significantly increased in atg5 KO MEF indicating that atg5 plays an important role in the maturation of autophagosomes. Next, we examined autophagosomes in the cell model expressing polyQ-expanded N-terminal fragment of huntingtin. Our TEM analysis indicates that the number of late autophagosomes was significantly increased in the cells expressing the mutant huntingtin, indicating that improving the fusion of autophagosome with lysosome may be effective to enhance autophagy for the treatment of Huntington's disease. Taken together, the results of our study indicate that ultrastructural and quantitative analysis of autophagosomes using TEM can be applied to various human cellular disease models, and that they will provide an important insight for cellular pathogenesis of human diseases associated with autophagy.

• Current Research on Agriculture and Life Sciences
• /
• v.12
• /
• pp.57-68
• /
• 1994

This experiment was conducted to isolate the mutants from S118 and to investigate the physiological characteristics of R. japonicum mutants. The results obtained were as follows; Based on nodulation and acetylene reduction, nodulation of rhizobia was divided into 4 groups, i.e. slow-nodulation, earlier-nodulation, infrequent-nodulation and non-nodulation. At 5% significant level, the growth of inoculated plant with SM255 was bad, but that of HP277 was good. Root-hairs curling was induced by strains S118 and HP277 on soybean, but not by strain SM255. S118 and SM255 were found to be slow-gorwers and produced alkali, whereas strain HP277 was fast-grower and produced acid in YEM broth. In litmus milk reaction, all strains indicated alkaline reaction, and serume-zone was induced weakly by HP277. All of the strains tested in this experiment utilized sucrose. HP277 and LP268 utilized xylose, whereas S118 and SM255 did not. SM255 showed bad growth in nitrogen carriers however utilization of $Ca(NO_3)_2{\cdot}4H_2O$ by HP277 was possible at 25mM and 10mM level. To compare with S118, the protein band of SM255's cell protein electrophoresis was not developed at 0.62 Rm position.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.31 no.3
• /
• pp.429-436
• /
• 1998

A bacteriological study of sea water and oyster in Charan Bay was conducted to evaluate sanitary conditions of the bay and compliance of waters with the recommended bacteriological criteria for the designated area of shellfish cultivation, The Samples were collected at 23 sampling stations(Fig. 1 and Fig. 2) estaslished once a month from January 1997 to December 1997, During the study period, temperature ranged from 4.7 to $25.6^{\circ}C$, transparency ranged from 3.3 to 6.2m chemical oxygen demand ranged from 1.67 to 2.18 mg/$\ell$, dissolved oxygen demand ranged from 5.4 to 10.0 mg/$\ell$ dissolved nitrogen ranged from 1.65 to 7.88 $\mu$g-at/$\ell$, phosphate ranged from 0.15 to 1.16 $\mu$g-at/$\ell$, Chlorophylla-a ranged from 0.95 to 12.69mg/$\ell$. The coliform group and fecal coliform MPN's of sea water were ranged from <1.8$\~$l,600 and <1.8$\~$540, respectively. The coliform group and fecal coliform MPN's of oysters were ranged from <18$\~$16,000 and <18$\~$1,400, respectively. The viable cell counts in oyster ranged from $1.5\times10^2$ to $7.5\times10^3$. The bacteriological criteria of sea water in shellfish growing area should be less than 70 per 100 ml of sea water for median value of coliform MPN, and below $10\%$ of the samples which contain over than 230 for coliform MPN or over than 43 for fecal coliform MPN. The sea water from 432 samples were complied water coliform criteria recommended for designated shellfish growing area. The coliform group, fecal coliform, classification of coliform group with IMViC reactions and pathogenic vibrios were analyzed. During the study period, infectious bacteria such as Vibrio cholerae, Salmonella sp, and Shigella sp, were not detected from the samples, but detection ratios of Vibrio parahaemolyticus and Vibrio vulnifirus were $7\~17\%$ in summer months.

• Current Research on Agriculture and Life Sciences
• /
• v.8
• /
• pp.45-50
• /
• 1990

This study was carried out ot investigate the effects of fetal calf serum (FCS) and gonadotropins supplemented to the medium on maturation and fertilization in vitro of porcine follcular oocytes. Ovaries were obtained from gilts at local slaughter-house. Oocyte-cumulus complexes were recovered by puncturing the ovarian follicles(3~5 mm in diameter). The complexes from individual ovaries were pooled in a $0.4m{\ell}$ droplet of medium covered with paraffin oil, then washed twice in fresh droplet and cultured for 36hrs in culture media according to experimental conditions. Boar epididymal spermatozoa were capacitated by preincubation for 4hrs in m-KRB medium and the preincubated spermatozoa were insemenated in the fertilization medium containing the cultured oocytes. The results obtained in this study are summarized as follows: 1. The maturation rates of oocytes cultured in m-KRB and m-KRB supplemented to 10% FCS were 82 and 37%, respectively. When PMSG, hCG. and PMSGt hcG($10Iu/m{\ell}$) were added to the media supplemented to 10% FCS, the maturation rates were 66, 58 and 68%, respectively. 2. Expansion of cumulus cells was not occured in m-KRB and m-KRB supplemented to 10% FCS. However, when PMSG, hCG and PMSG+hCG($10Iu/m{\ell}$) were added to m-KRB supplemented to 10% FCS, the expansion rates of cumulus cell layers were 92, 13 and 91%, respectively. 3. When oocytes were mltured in m-KRB, the rates of penetration and formation of male pronucle: were 93 and 7%, respectively. By adding FCS and gonadotropin to m-KRB, the penetration and formation of male pronuclei were 100 80%, respectively.

• Journal of Korean Society of Environmental Engineers
• /
• v.39 no.10
• /
• pp.591-598
• /
• 2017

Power overshoot phenomenon was observed in microbial fuel cells (MFCs) used non-catalyzed graphite felt as cathode. Voltage loss in MFCs was mainly caused by cathode potential loss. Cheap stainless steel scrubber, which has high conductivity, and Pt/C coated graphite felt as cathode were used for overcoming power overshoot and reducing the cathode potential loss in MFCs. The MFCs used stainless steel scrubber showed no power overshoot even slow catholyte flow rate and produced 29% enhanced maximum current density ($23.9A/m^3$) than MFCs used non-catalyzed graphite felt while the power overshoot phenomenon was existed in Pt/C coated MFCs. Increasing catholyte flow rate resulted in disappearing power overshoot of MFCs used non-catalyzed graphite felt. In addition, maximum power density and current density of both MFCs used non-catalyzed graphite felt and stainless steel scrubber increased by 2-3.5 times. Cathode potential losses in all region of activation loss, ohmic loss, and mass transport loss were reduced according to increase of catholyte flow rate. Therefore, stainless steel scrubber has advantages that are economical materials as electrode and prevents power overshoot, leading to enhance electricity generation. In addition, increasing catholyte flux is one of great solution when power overshoot caused by cathodic overpotential is observed in MFCs.

• Korean Security Journal
• /
• no.9
• /
• pp.99-131
• /
• 2005

Terrorism which became today's common phenomena over the world is one of the most serious threats the world confront. Although International society make and operate outstanding anti-terrorism system, terror would never end without solving fundamental problems. The main body of terrorism converts from nation to organization and from organization to cell, which makes it difficult for us to recognize the main body. Since the target of today's new terrorism is many and unspecified persons, terrorists will never hesitate to use mass destruction weapons such as nuclear, biological, chemical weapons, and also use cyber-technique or cyber-terrorism. So, effective counter-terrorism measures should be performed as follows. First, it must be better for international society should make long-time plan of solving fundamental problems of terrorism other than to operate directly on terror organization and its means. Second, preventive method should be made. The most effective method of eradicating terrorism is prevention. For this, it is necessary to remove environmental elements of terrorism and terrorist bases, and to stop inflow of money and mass destruction weapons to terrorists. Third, integrated anti-terror organization should be organized and operated for continuous counter-terrorism operations. Also international alliance for anti-terrorism should be maintained to share informations and measures. Fourth, concerned department in the government should prepare counter-terrorism plans in their own parts as follows and make efforts to integrate the plans. - Ministry of Government Administration and Home Affairs : conventional terror - Ministry of Health and Welfare : bio-terror - Ministry of Science and Technology : nuclear-terror Especially, they should convert their policy and operation from post-terror actions to pre-terror actions, designate terror as national disaster and organize integrated emergency response organization including civil, government, and military elements. In conclusion, pre-terror activities and remedy of fundamental causes is the best way to prevent terror. Also, strengthening of intelligence activities, international cooperations, and preventive and comprehensive counter-measures must not ignored.

• Journal of the Korean Crystal Growth and Crystal Technology
• /
• v.24 no.6
• /
• pp.229-236
• /
• 2014

A stoichiometric mixture of evaporating materials for $MnAl_2S_4$ single crystal thin films was prepared from horizontal electric furnace. To obtain the single crystal thin films, $MnAl_2S_4$ mixed crystal was deposited on thoroughly etched semi-insulating GaAs(100) substrate by the Hot Wall Epitaxy (HWE) system. The source and substrate temperatures were $630^{\circ}C$ and $410^{\circ}C$, respectively. The crystalline structure of the single crystal thin films was investigated by the photoluminescence and double crystal X-ray diffraction (DCXD). The temperature dependence of the energy band gap of the $MnAl_2S_4$ obtained from the absorption spectra was well described by the Varshni's relation, $E_g(T)=3.7920eV-5.2729{\times}10^{-4}eV/K)T^2/(T+786 K)$. In order to explore the applicability as a photoconductive cell, we measured the sensitivity (${\gamma}$), the ratio of photocurrent to dark current (pc/dc), maximum allowable power dissipation (MAPD) and response time. The results indicated that the photoconductive characteristic were the best for the samples annealed in S vapour compare with in Mn, Al, air and vacuum vapour. Then we obtained the sensitivity of 0.93, the value of pc/dc of $1.10{\times}10^7$, the MAPD of 316 mW, and the rise and decay time of 14.8 ms and 12.1 ms, respectively.

• Journal of the Korean Electrochemical Society
• /
• v.15 no.3
• /
• pp.172-180
• /
• 2012

The $LiMn_2O_4$ cathodes for lithium ion battery were synthesized from various precursors of manganese oxides and manganese hydroxides. As the first step, nanosized precursors such as ${\alpha}-MnO_2$ (nano-sticks), ${\beta}-MnO_2$ (nano-rods), $Mn_3O_4$ (nano-octahedra), amorphous $MnO_2$(nano-spheres), and $Mn(OH)_2$ (nano-plates) were prepared by a hydrothermal or a precipitation method. Spinel $LiMn_2O_4$ with various sizes and shapes were finally synthesized by a solid-state reaction method from the manganese precursors and LiOH. Nano-sized (500 nm) octahedron $LiMn_2O_4$ showed high capacities of 107 mAh $g^{-1}$ and 99 mAh $g^{-1}$ at 1 C- and 50 C-rate, respectively. Three dimensional octahedral crystallites exhibit superior electrochemical characteristics to the other one-dimensional and two-dimensional shaped $LiMn_2O_4$ nanoparticles. After 500 consecutive charge discharge battery cycles at 10 C-rate with the nano-octahedron $LiMn_2O_4$ cathode, the capacity retention of 95% was observed, which is far better than any other morphologies studied in this work.

• Journal of Life Science
• /
• v.28 no.4
• /
• pp.389-397
• /
• 2018

The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

• Journal of Life Science
• /
• v.30 no.8
• /
• pp.701-707
• /
• 2020

Gene editing technology using the clustered regularly interspaced short palindromic repeat (CRISPR) and the CRISPR associated protein (Cas)9 has been highly anticipated in developing breeding techniques. In this study, we discuss gene scissors as a tool for silkworm molecular breeding through analysis of Bombyx mori Kynurenine 3-Monooxygenase (BmKMO) gene editing using the CRISPR/Cas9 system and analysis of generational transmission through mutagenesis and selective crossing. The nucleotide sequence of the BmKMO gene was analyzed, and three guide RNAs (gRNAs) were prepared. Each synthesized gRNA was combined with Cas9 protein and then analyzed by T7 endonuclease I after introduction into the BM-N silkworm cell line. To edit the silkworm gene, K1P gRNA and Cas9 complexes were subsequently microinjected into the silkworm embryos; the hatching rate was 18% and the incidence of mutation was 60%. The gene mutation was verified in the heterozygous G0 generation, but no phenotypic change was observed. In homozygotes generated by self-crossing, a mutant phenotype was observed. These results suggest that silkworm molecular breeding using the CRISPR/Cas9 system is possible and could be an effective way of shortening the time required.