• Journal of Microbiology
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• v.45 no.2
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• pp.158-167
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• 2007

In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

• Bulletin of the Korean Chemical Society
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• v.35 no.9
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• pp.2615-2620
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• 2014

Ni/$La_2O_3$ with a high dispersion was prepared by reduction of $La_2O_3$ perovskite oxide to examine the catalytic activity for the $CO_2-CH_4$ reaction. The Ni/$La_2O_3$ catalyst was found to be highly active for the reaction. The ratios of $H_2$/CO were measured in a flow of the reaction mixture containing $CO_2/CH_4$/Ar using an on-line gas chromatography system operated at 1 atm and found to be varied with temperature between 0.66 and 1 in the temperature range of $500-800^{\circ}C$. A kinetic study of the catalytic reaction was performed in a static reactor at 40 Torr total pressure of $CO_2/CH_4/N_2$ by using a photoacoustic spectroscopy technique. The $CO_2$ photoacoustic signal varying with the concentration of $CO_2$ during the catalytic reaction was recorded as a function of time. Rates of $CO_2$ disappearance in the temperature range of $550-700^{\circ}C$ were obtained from the changes in the $CO_2$ photoacoustic signal at early reaction stage. The plot of ln rate vs. 1/T showed linear lines below and above $610^{\circ}C$. Apparent activation energies were determined to be 10.4 kcal/mol in the temperature range of $550-610^{\circ}C$ and 14.6 kcal/mol in the temperature range of $610-700^{\circ}C$. From the initial rates measured at $640^{\circ}C$ under various partial pressures of $CO_2$ and $CH_4$, the reaction orders were determined to be 0.43 with respect to $CO_2$ and 0.33 with respect to $CH_4$. The kinetic results were compared with those reported previously and used to infer a reaction mechanism for the Ni/$La_2O_3$-catalyzed $CO_2-CH_4$ reaction.

• The Korean Journal of Food And Nutrition
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• v.22 no.4
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• pp.625-632
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• 2009

This study determined the effects of added buckwheat extract on the rate of corn starch hydrolysis in vitro as well as blood glucose responses through its supplementation in healthy subjects. The rate of corn starch hydrolysis in the presence or absence of various buckwheat extracts was determined in an in vitro enzyme/dialysis system for 2 hr. The buckwheat was extracted by water, ethanol(40%, 70%, 100%) and methanol(40%, 70%, 100%), respectively. Twenty percent(w/w) additions of the ethanol, methanol and water buckwheat extract to corn starch solution significantly reduced the starch hydrolysis at every minute for 2 hr(p<0.05). The calculated hydrolysis indices of the buckwheat extracts were in the order of 100% ethanol extract(50), 100% methanol(54), 40% ethanol(58), 40% methanol(62), 70% methanol(64), 70% ethanol(68), water (82). For the blood glucose response study, groups of 12 volunteers were given 50 g of boiled rice with or without buckwheat extract(10% and 20% of starch weight) using the 100%, 70%, and 40% ethanol extracts, respectively. The addition of each buckwheat ethanol extract significantly reduced blood glucose concentrations at three or more points during 2 hr and also reduced the mean peak rise and area under the blood glucose curve(p<0.05). The calculated glycemic index(GI) values for all ethanol buckwheat extract groups were significantly decreased compared to the control(rice). At the concentrations of 20%, the buckwheat 100% ethanol extracts lowered the GI by 68%. The 100% ethanol extract was more effective than the 70% and 40% extracts for reducing GI. Therefore, the 100% ethanol buckwheat extract would be the most therapeutically useful in modifying postprandial hyperglycemia.

• BMB Reports
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• v.39 no.2
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• pp.197-207
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• 2006

Cajanus indicus is a herb with medicinal properties and is traditionally used to treat various forms of liver disorders. Present study aimed to evaluate the effect of a 43 kD protein isolated from the leaves of this herb against chloroform induced hepatotoxicity. Male albino mice were intraperitoneally treated with 2mg/kg body weight of the protein for 5 days followed by oral application of chloroform (0.75ml/kg body weight) for 2 days. Different biochemical parameters related to physiology and pathophysiology of liver, such as, serum glutamate pyruvate transaminase and alkaline phosphatase were determined in the murine sera under various experimental conditions. Direct antioxidant role of the protein was also determined from its reaction with Diphenyl picryl hydraxyl radical, superoxide radical and hydrogen peroxide. To find out the mode of action of this protein against chloroform induced liver damage, levels of antioxidant enzymes catalase, superoxide dismutase and glutathione-S-transferase were measured from liver homogenates. Peroxidation of membrane lipids both in vivo and in vitro were also measured as malonaldialdehyde. Finally, histopathological analyses were done from liver sections of control, toxin treated and protein pre- and post-treated (along with the toxin) mice. Levels of serum glutamate pyruvate transaminase and alkaline phosphatase, which showed an elevation in chloroform induced hepatic damage, were brought down near to the normal levels with the protein pretreatment. On the contrary, the levels of anti-oxidant enzymes such as catalase, superoxide dismutase and glutathione-S-transferase that had gone down in mice orally fed with chloroform were significantly elevated in protein pretreated ones. Besides, chloroform induced lipid peroxidation was effectively reduced by protein treatment both in vivo and in vitro. In cell free system the protein effectively quenched diphenyl picryl hydrazyl radical and superoxide radical, though it could not catalyse the breakdown of hydrogen peroxide. Post treatment with the protein for 3 days after 2 days of chloroform administration showed similar results. Histopathological studies indicated that chloroform induced extensive tissue damage was less severe in the mice livers treated with the 43 kD protein prior and post to the toxin administration. Results from all these data suggest that the protein possesses both preventive and curative role against chloroform induced hepatotoxicity and probably acts by an anti-oxidative defense mechanism.

• Journal of Korean Tunnelling and Underground Space Association
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• v.20 no.1
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• pp.161-182
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• 2018

We carried out a series of linear cutting machine tests to assess the cutting performance of a pick cutter in sedimentary rock. The specimens were Linyi sandstone from China and Concrete (rock-like material, conglomerate). Using the small scaled LCM system, we estimated the cutter force and specific energy under different cutting conditions. The cutter forces (cutting and normal) increased with penetration depth and cutter spacing in two rock types, and it was affected by the strength of specimens. On the other hand, the ratio of the peak cutter force to the mean cutter force was influenced by cutting characteristic and composition of rock rather than rock strength. The cutting coefficient was affected by the friction characteristic between rock and pick cutter rather than the cutting conditions. Therefore, the optimal cutting angle can be determined by considering of cutting coefficient and resultant force of pick cutter. The optimum cutting condition was determined from the relationship between the specific energy and cutting condition. For two specimens, the optimum s/p ratio was found to be two to four, and the specific energy decreased with the penetration depth. The result from this study can be used as background database to understand the cutting mechanism of a pick cutter, also it can be used to design for the mechanical excavator.

• Toxicological Research
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• v.33 no.3
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• pp.239-253
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• 2017

Neodymium is a future-oriented material due to its unique properties, and its use is increasing in various industrial fields worldwide. However, the toxicity caused by repeated exposure to this metal has not been studied in detail thus far. The present study was carried out to investigate the potential inhalation toxicity of nano-sized neodymium oxide ($Nd_2O_3$) following a 28-day repeated inhalation exposure in male Sprague-Dawley rats. Male rats were exposed to nano-sized $Nd_2O_3-containing$ aerosols via a nose-only inhalation system at doses of $0mg/m^3$, $0.5mg/m^3$, $2.5mg/m^3$, and $10mg/m^3$ for 6 hr/day, 5 days/week over a 28-day period, followed by a 28-day recovery period. During the experimental period, clinical signs, body weight, hematologic parameters, serum biochemical parameters, necropsy findings, organ weight, and histopathological findings were examined; neodymium distribution in the major organs and blood, bronchoalveolar lavage fluid (BALF), and oxidative stress in lung tissues were analyzed. Most of the neodymium was found to be deposited in lung tissues, showing a dose-dependent relationship. Infiltration of inflammatory cells and pulmonary alveolar proteinosis (PAP) were the main observations of lung histopathology. Infiltration of inflammatory cells was observed in the $2.5mg/m^3$ and higher dose treatment groups. PAP was observed in all treatment groups accompanied by an increase in lung weight, but was observed to a lesser extent in the $0.5mg/m^3$ treatment group. In BALF analysis, total cell counts, including macrophages and neutrophils, lactate dehydrogenase, albumin, interleukin-6, and tumor necrosis factor-alpha, increased significantly in all treatment groups. After a 4-week recovery period, these changes were generally reversed in the $0.5mg/m^3$ group, but were exacerbated in the $10mg/m^3$ group. The lowest-observed-adverse-effect concentration of nano-sized $Nd_2O_3$ was determined to be $0.5mg/m^3$, and the target organ was determined to be the lung, under the present experimental conditions in male rats.

• Journal of the Korean Society of Marine Environment & Safety
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• v.24 no.2
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• pp.157-162
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• 2018

The increase of risk in port due to the increase in ship size and sea level rises, the standard crown height will increase. In this study, cruise and container ships will need to raise their crown height due to the projected wind pressure areas becoming larger due to the ships' size increase. The mooring assessment was evaluated with the rise of the crown height. The cruise ship of GT 100,000 tons exceeded the permissible breaking force of the mooring line under the crown height conditions of wind speed of 30 kts when the wind direction was $45^{\circ}$ to the direction of the bow. Also, the elevation angle of the pier and mooring line was analyzed and exceeded the crown height, and it was determined that it is necessary to adjust the crown height. Container ships of DWT 100,000 tons were analyzed to exceed the limit of sway motion at the crown height and it was determined that they need to be adjusted to the minimum crown height standard.

• Weed & Turfgrass Science
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• v.4 no.4
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• pp.308-314
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• 2015

This study was conducted to establish a fluorescence assay system for high efficient mass screening of the herbicides causing rapid membrane peroxidation, based on the fact that peroxide in cellular leakage could be fluorometrically determined through the fuorescent compounds formed after reacting with homovanillic acid (HVA) and peroxidase (HRP). The assay procesure established in this study was as follows. Only single disc (4 mm diameter) excised from cucumber cotyledon is placed on the well containing test solution ($200{\mu}L$) with 96-well microplate. The plate is shaking-incubated for 8 h under light condition. Then after removing the cucumber disc, HVA and HRP are supplied in the medium buffer and incubated for 5 min at room temperature. Fluorescence values are determined at Ex 320 nm/Ex 425 nm. The higher fluorescence values are obtained in the treatment of chemical having higher herbicidal activity. Using this assay with 96-well microplates, a large number of herbicides inducing rapid membrane peroxidation seemed to be screened more efficiently than spectrophotometric microtiter assay reported previously.

• Journal of Climate Change Research
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• v.9 no.2
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• pp.181-195
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• 2018

On December 12, 2015, the Conference of the Parties to the United Nations Framework Convention on Climate Change (UNFCCC) adopted the Paris Agreement, in which several developed and developing countries all committed to participating in the reduction of greenhouse-gas (GHG) emissions. South Korea has submitted an intended nationally determined contribution (INDC) proposal with a target to cut down 37% greenhouse gas business as usual (BAU) until 2030 in preparation for the 2030 GHG BAU. Under the post-2020 regime, which will be launched from 2021 as the agreement entered into force early, it is expected that efforts to support GHG reduction and adaptation to climate change in developing countries will be accelerated with the utilization of technologies and financial resources of developed countries. South Korea has established the Basic Plan for Climate Change Response and the Basic National Roadmap for Greenhouse Gas Reductions by 2030 to promote the response to climate change at the government level. The Ministry of Science and ICT, as the National Designated Entity designated by the UNFCCC, has come up with middle and long-term strategies for climate technology cooperation. South-Korea has an abundance of energy-consuming industries to support its export-oriented industrial structure; it is thus expected that achieving the GHG reduction target will incur a considerable cost. Moreover, in order to meet the reduction target (11.3%) of the intended nationally determined contribution proposed by South Korea, it is necessary for South Korea to actively promote projects that can achieve GHG reduction achievements, and financial resources are needed as leverage to reduce risks that can occur in the early stages of projects and attract private sector investment. This paper summarizes the theoretical discussions on climate finance and conducted a comparative analysis on the status of the funds related to climate change response in the UK, Germany, Japan and Denmark. Through this, we proposed the legal and policy tasks that should be carried forward to raise public funds that can be used for creation of new industries related to climate change as well as to reduce GHG emissions in South Korea. The Climate Change Countermeasures Act, which has been proposed by the National Assembly of South-Korea, stipulates the establishment of funds but there is no additional funding except for general account. In this regard, it is also possible to take measures such as the introduction of carbon tax or the collection and use of royalties through technology research and development projects for climate change, such as Industrial Technology Innovation Promotion Act. In addition, since funds are used in various fields such as domestic greenhouse gas reduction, technology development, and overseas projects, it is necessary to establish a system in which various ministries cooperate with the operation of the fund.

• Journal of the Korean Society of Marine Environment & Safety
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• v.27 no.2
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• pp.228-237
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• 2021

In those day, the Busan local government had instituted a rule to support and enhance competition as well as improve respect for the ship management industry. This study aims to extract the cause factors to enhance such competition using intuitionistic decision making trial and evaluation laboratory (DEMATEL) and interpretive structural modeling (ISM) methods. First, eight factors were extracted from the specifications in the Ship Management Industry Development Act. Second, the intuitionistic fuzzy number was converted to a crisp number using the standard fuzzy number. Third, the influence relationship was analyzed using DEMATEL, and the priority ranks for the factors are determined using ISM. From the results of the impact relationship analysis, the three main cause factors were determined as improvement of technical ship management capability, improvement of expertise of manpower for onshore management, and improvement of the quality of the Korean seafarer. The priorities under the ISM method, in descending order, were as follows: improvement of the quality of Korean seafarers, improvement of professionalism among the manpower for shore management, improvement of technical ship management capability, improvement of commercial ship management capability, establishment of a comprehensive information system, improvement of the working conditions and employment environment for seafarers, financial support such as overseas orders, and strengthening the availability of foreign seafarers. Therefore, it is necessary to prioritize policy promotion based on these factors, especially the top three, as these have the highest impact.