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Immunogenicity of Synthetic Peptide Specific for Major Immunogenic Determinat of Hepatitis B Surface Antigen (B형간염(型肝炎) 표면항원(表面抗原)의 주면역원(主免疫原) 결정기(決定基)에 특이(特異)한 합성(合成) Peptide의 면역원성(免疫原性)에 관한 연구(硏究))

  • Shin, Kwang-soon;Han, Su-nam
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.7-17
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    • 1985
  • Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

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Improvement of an Simultaneous Determination for Clenbuterol and Ractopamine in Livestock Products using LC-MS/MS (LC-MS/MS를 이용한 축산물 중 clenbuterol과 ractopamine의 동시 분석법 개선)

  • Cho, Yoon-Jae;Chae, Young-Sik;Kim, Jae-Young;Kim, Hyung-Soo;Kang, Ilhyun;Do, Jung-Ah;Oh, Jae-Ho;Kwon, Kisung;Chang, Moon-Ik
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.25-33
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    • 2013
  • Clenbuterol and ractopamine, which are ${\beta}$-agonists, have been misused as a growth promoting agent in meat producing animals. Clenbuterol was banned for veterinary drug in Korea because of its problems regarding safety. Due to their adverse effects, such as cardiovascular and central nervous diseases on human health proper control and monitoring should be conducted. The existing analytical method of clenbuterol and ractopamine in the Food code was improved through our present study. The bovine muscle samples were subjected to enzymatic hydrolysis, extracted with ethyl acetate and defatted by hexane-methanol partitioning. A molecular imprinted polymer (MIP) solid phase extraction cartridge was used for clean-up and LC-MS/MS was operated in positive multiple reaction monitoring (MRM). Clenbuterol-$d_9$ and ractopamine-$d_3$ were used as an internal standard. The renewed method was validated according to the CODEX guideline. The limits of quantitation for clenbuterol and ractopamine were 0.2 and 0.5 ${\mu}g/kg$, respectively. The mean recoveries ranged in 104.2-113.5% for clenbuterol and in 107.6-118.1% for ractopamine. The improved method was able to save both time and expenses.

Laboratory Tests for Trichloroethylene (TCE) and Toluene Remediation in Soil Using Soil Vapor Extraction (토양증기추출(Soil Vapor Extraction)을 이용한 토양 내 Trichloroethylene (TCE)과 Toluene정화 실험)

  • 이민희;강현민
    • Economic and Environmental Geology
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    • v.35 no.3
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    • pp.221-227
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    • 2002
  • Column experiments were performed to evaluate the removal efficiency of soil vapor extraction (SVE) iota TCE (trichloroethylene) and toluene in soil. Homogeneous Ottawa sands and real soils collected from contaminated area were used to investigate the effect of soil properties and SVE operation conditions on the removal efficiency. In column teats with two different sizes of Ottawa sand, the maximum effluent TCE concentration in a coarse sand column was 442 mg/L and 337 mg/L in a fine sand column. However, after 20 liter gas flushing, the effluent concentrations were very similar and more than 90% of initial TCE mass were removed from the column. For two real contaminated soil columns, the maximum effluent concentration decreased 50% compared with that in the homogeneous Ottawa coarse sand column, but 99% of initial TCE mass were extracted from the column within 40 liter air flushing, suggesting that SVE is very available to remove volatile NAPLs in the contaminated soil. To investigate the effect of contaminant existing time on the removal efficiency, an Ottawa sand column was left stable for one week after TCE was injected and the gas extraction was applied into the column. Its effluent concentration trend was very similar to those for other Ottawa sand columns except that the residual TCE after the air flushing showed relatively high. Column tests with different water contents were performed and results showed high removal efficiency even in a high water content sand column. Toluene as one of BTEX compounds was used in an Ottawa sand column and a real soil column. Removal trends were similar to those in TCE contaminated columns and more than 98% of initial toluene mass were removed with SVE in both column.

Isolation and Characterization of a Marine Bacterium, Pseudomonas sp. YJ-1 with Anti-Methicillin Resistant Staphylococcus aureus Activity (항 Methicillin Resistant Staphylococcus aureus 활성을 가지는 해양미생물 Pseudomonas sp. YJ-1의 분리와 특성)

  • Woo, Ye-Ju;Jeong, Seong-Yun
    • Korean Journal of Environmental Biology
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    • v.35 no.4
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    • pp.694-705
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    • 2017
  • The aim of this study was to isolate and identify marine bacterium with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were $25^{\circ}C$ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a $C_{18}$ ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of $250{\mu}g\;mL^{-1}$ concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.

Identification of Fatty Acids in the Pulp Oils of Jujube and Their Compsitional Changes in the Ripening Period (대추의 과육지질(果肉脂質)에 존재(存在)하는 지방산(脂肪酸)의 동정(同定)과 숙성(熟成)에 따른 그 조성(組成)의 변화(變化))

  • Woo, Hyo-Kyeng;Kim, Seong-Jin;Park, Sung-Hea;Joh, Yong-Goe
    • Journal of the Korean Applied Science and Technology
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    • v.18 no.1
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    • pp.67-77
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    • 2001
  • In search for several fatty acid with unusual structure in vegetable oils, we have found that unknown peaks were shown on GLC in the analysis of fatty acids of the lipids from the pulp of ripened jujube (Zizypus jujuba var. inermis) fruits. These fatty acids were identified as a series of cis-monoenoic acids with ${\omega}-5$ double bond system such as $C_{14:1{\omega}5}$, $C_{16:1{\omega}5}$ and $C_{18:1{\omega}5}$, including ${\omega}-7$ fatty acid as $C_{16:1{\omega}7}$ and $C_{18:1{\omega}7}$, by GLC, solid-phase extraction silver ion-column chromatographic, GLC-mass spectrometric and IR techniques. First of all, total fatty acid methyl esters were resolved into saturated and branched fatty acid, monoenoic acid, dienoic acid, and trienoic acid fraction, respectively, with 100% dichloromethane (DCM), DCM/acetone (9:1, v/v) 100% acetone, and acetone/ acetonitrile (97:3, v/v) solvent system. Unknown fatty acids were included in the monoenoic fraction and were confirmed to have cis-configuration by IR. Picolinyl esters of monoenoic fatty acids gave distinct molecular ion peak and dominant diagnostic peaks, for example, m/z 317, 220 and 260 fragment for $cis-C_{14:1{\omega}5}$, m/z 345, m/z 248 and 288 fragment for $cis-C_{16:1{\omega}5}$ and m/z 373, m/z 276 and 316 fragment for $cis-C_{18:1{\omega}5}$. In this way the occurrence of $cis-C_{16:1{\omega}7}$ and $cis-C_{18:1{\omega}7}$ could be deduced from the appearance of prominent fragments as m/z 345, 220 and 260, and m/z 373, 248 and 280. Level of total ${\omega}-5$ fatty acids amounted to about 30% in the fatty acid composition with the predominance of $C_{16:1{\omega}5}$ $ (18.7{\sim}25.0%)$, in the semi-ripened and/or ripened samples collected in September 14 ($C_{16:1{\omega}5}$ ; 18.7%, $C_{14:1{\omega}5}$ ; 3.6% and $C_{18:1{\omega}5}$ ; 3.0%), September 22 ($C_{16:1{\omega}5}$ ; 25.0%, $C_{14:1{\omega}5}$ ; 1.4% and $C_{18:1{\omega}5}$ ; 2.6%), and October $7 (C_{16:1{\omega}5}$ ; 24.7%, $C_{14:1{\omega}5}$ ; 7.7% and $C_{18:1{\omega}5}$ ; 2.5%). However, the lipids extracted from unripened jujube in July and August contain these unusual fatty acids as low as negligible. It could be observed that the level of ${\omega}-5$ fatty acids in the pulps increased sharply with an elapse of ripening time of jujube fruits. Other monoenoic fatty acids with ${\omega}-7$ series, $C_{16:1{\omega}7}$ (palmitoleic acid) and $C_{18:1{\omega}7}$ (cis-vaccenic acid) could be detected. And in the lipids of the kernel and leaf of jujube, none of ${\omega}-5$ fatty acids could be detected.

Channel Innovation through Online Transaction processing System in Floral Wholesale Distribution: FLOMARKET Case (화훼도매 온라인 거래처리 시스템을 통한 유통경로 개선방안 연구: (주)플로마켓 사례)

  • Lee, Seungchang;Ahn, Sunghyuck
    • Journal of Distribution Science
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    • v.8 no.1
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    • pp.21-33
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    • 2010
  • The ICT(information & communication technology) led to a dramatic change of floral distribution service, a phase of competition between wholesales and retail stores, and distribution channels in floral industry. It was expected that a role of the intermediaries in this industry would have reduced due to the improvement of transaction process by ICT. However, the ICT made to overcome a regional limit of the floral retail distribution service leading to an increase in sales and enlargement of the stores. And even it made possible to bring out another type of intermediaries such as private associations. This case study focuses on what kinds of efforts the floral wholesale distributors have made to enable a distribution process more smoothly between the wholesale distributors and retail stores through the information system, and what the failure factors in adopting the information system have been. This paper is also to examine how the wholesale distributors have changed themselves to gain dominant positions in distribution channels. As a result of the study, it was found that the intermediaries mostly failed in successfully achieving the distribution channel innovation through the information system because of several main reasons. FLOMARKET Inc. tried to innovate a distribution channel to obtain high quality goods through consolidating a wholesale distribution market in that segregated both floral joint market from free markets. after implementing the information system with consideration of the failure factors, FLOMARKET Inc. was able to minimize goods in stock and make a major purchase of various goods. In addition, it made a possible pre-ordering process and an exact calculation of purchasing goods so they could provide their products with market price in real time, which helped for the company to gain credits from their customers. Also, FLOMARKET Inc. established the information system which well suited to its business stage in order to deal with a rapidly changing distribution environment. It's so obvious that the transaction processing system of FLOMARKET Inc. definitely helped to share information among traders more seamlessly and smoothly in realtime, standardize goods, and make a transaction process clearer. Besides, the transaction information helped the wholesale distributors and retail stores to make more strategic decisions in their business because through the system they enabled to gather the marketing intelligence information more easily and convenient. If we understand that the floral distribution market is characterized by the low IT- based industry, it's worth to examine a case study proving that the information system actually increases the productivity of the transaction process in the floral industry.

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Effects of acid-treatment conditions on the surface properties of the RBM treated titanium implants (산-처리 조건이 RBM처리한 티타늄 임플란트의 표면 특성에 주는 영향)

  • Lee, Han-Ah;Seok, Soohwang;Lee, Sang-Hyeok;Lim, Bum-Soon
    • Korean Journal of Dental Materials
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    • v.45 no.4
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    • pp.257-274
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    • 2018
  • The purpose of this study was to evaluate the effect of acid-treatment conditions on the surface properties of the RBM (Resorbable Blast Media) treated titanium. Disk typed cp-titanium specimens were prepared and RBM treatments was performed with calcium phosphate ceramic powder. Acid solution was mixed using HCl, $H_2SO_4$ and deionized water with 4 different volume fraction. The RBM treated titanium was acid treated with different acid solutions at 3 different temperatures and for 3 different periods. After acid-treatments, samples were cleaned with 1 % Solujet solution for 30 min and deionized water for 30 min using ultrasonic cleanser, then dried in the electrical oven ($37^{\circ}C$). Weight of samples before and after acid-treatment were measured using electric balance. Surface roughness was estimated using a confocal laser scanning microscopy, crystal phase in the surface of sample was analyzed using X-ray diffractometer. Surface morphology and components were evaluated using Scanning Electron Microscope (SEM) with Energy Dispersive X-ray spectroscopy (EDX) and X-ray Photoemission Spectroscopy (XPS). Values of the weight changes and surface roughness were statistically analyzed using Tukey-multiple comparison test (p=0.05). Weight change after acid treatments were significantly increased with increasing the concentration of $H_2SO_4$ and temperature of acid-solution. Acid-treatment conditions (concentration of $H_2SO_4$, temperature and time) did not produce consistent effects on the surface roughness, it showed the scattered results. From XRD analysis, formation of titanium hydrides in the titanium surface were observed in all specimens treated with acid-solutions. From XPS analysis, thin titanium oxide layer in the acid-treated specimens could be evaluated. Acid solution with $90^{\circ}C$ showed the strong effect on the titanium surface, it should be treated with caution to avoid the over-etching process.

Recent research activities on hybrid rocket in Japan

  • Harunori, Nagata
    • Proceedings of the Korean Society of Propulsion Engineers Conference
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    • 2011.04a
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    • pp.1-2
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    • 2011
  • Hybrid rockets have lately attracted attention as a strong candidate of small, low cost, safe and reliable launch vehicles. A significant topic is that the first commercially sponsored space ship, SpaceShipOne vehicle chose a hybrid rocket. The main factors for the choice were safety of operation, system cost, quick turnaround, and thrust termination. In Japan, five universities including Hokkaido University and three private companies organized "Hybrid Rocket Research Group" from 1998 to 2002. Their main purpose was to downsize the cost and scale of rocket experiments. In 2002, UNISEC (University Space Engineering Consortium) and HASTIC (Hokkaido Aerospace Science and Technology Incubation Center) took over the educational and R&D rocket activities respectively and the research group dissolved. In 2008, JAXA/ISAS and eleven universities formed "Hybrid Rocket Research Working Group" as a subcommittee of the Steering Committee for Space Engineering in ISAS. Their goal is to demonstrate technical feasibility of lowcost and high frequency launches of nano/micro satellites into sun-synchronous orbits. Hybrid rockets use a combination of solid and liquid propellants. Usually the fuel is in a solid phase. A serious problem of hybrid rockets is the low regression rate of the solid fuel. In single port hybrids the low regression rate below 1 mm/s causes large L/D exceeding a hundred and small fuel loading ratio falling below 0.3. Multi-port hybrids are a typical solution to solve this problem. However, this solution is not the mainstream in Japan. Another approach is to use high regression rate fuels. For example, a fuel regression rate of 4 mm/s decreases L/D to around 10 and increases the loading ratio to around 0.75. Liquefying fuels such as paraffins are strong candidates for high regression fuels and subject of active research in Japan too. Nakagawa et al. in Tokai University employed EVA (Ethylene Vinyl Acetate) to modify viscosity of paraffin based fuels and investigated the effect of viscosity on regression rates. Wada et al. in Akita University employed LTP (Low melting ThermoPlastic) as another candidate of liquefying fuels and demonstrated high regression rates comparable to paraffin fuels. Hori et al. in JAXA/ISAS employed glycidylazide-poly(ethylene glycol) (GAP-PEG) copolymers as high regression rate fuels and modified the combustion characteristics by changing the PEG mixing ratio. Regression rate improvement by changing internal ballistics is another stream of research. The author proposed a new fuel configuration named "CAMUI" in 1998. CAMUI comes from an abbreviation of "cascaded multistage impinging-jet" meaning the distinctive flow field. A CAMUI type fuel grain consists of several cylindrical fuel blocks with two ports in axial direction. The port alignment shifts 90 degrees with each other to make jets out of ports impinge on the upstream end face of the downstream fuel block, resulting in intense heat transfer to the fuel. Yuasa et al. in Tokyo Metropolitan University employed swirling injection method and improved regression rates more than three times higher. However, regression rate distribution along the axis is not uniform due to the decay of the swirl strength. Aso et al. in Kyushu University employed multi-swirl injection to solve this problem. Combinations of swirling injection and paraffin based fuel have been tried and some results show very high regression rates exceeding ten times of conventional one. High fuel regression rates by new fuel, new internal ballistics, or combination of them require faster fuel-oxidizer mixing to maintain combustion efficiency. Nakagawa et al. succeeded to improve combustion efficiency of a paraffin-based fuel from 77% to 96% by a baffle plate. Another effective approach some researchers are trying is to use an aft-chamber to increase residence time. Better understanding of the new flow fields is necessary to reveal basic mechanisms of regression enhancement. Yuasa et al. visualized the combustion field in a swirling injection type motor. Nakagawa et al. observed boundary layer combustion of wax-based fuels. To understand detailed flow structures in swirling flow type hybrids, Sawada et al. (Tohoku Univ.), Teramoto et al. (Univ. of Tokyo), Shimada et al. (ISAS), and Tsuboi et al. (Kyushu Inst. Tech.) are trying to simulate the flow field numerically. Main challenges are turbulent reaction, stiffness due to low Mach number flow, fuel regression model, and other non-steady phenomena. Oshima et al. in Hokkaido University simulated CAMUI type flow fields and discussed correspondence relation between regression distribution of a burning surface and the vortex structure over the surface.

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A Study on Documentation Strategy for Archiving Locality (지역 아카이빙을 위한 기록화방안 연구)

  • Kwon, Soon-Myung;Lee, Seung-Hwi
    • The Korean Journal of Archival Studies
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    • no.21
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    • pp.41-84
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    • 2009
  • Lots of cultures, memories, histories of the local life have disappeared. Some sectors of universities and religion have keep their records in manuscript archive only. On the other hand records of public sectors were at least able to be managed by the records management law. Citizen's groups and academic bounds were also roles to get public records strong. However can we just describe whole body with only public records? As records management law a record of private sector which has value of preserving can be managed under national protection. Yet establishment of local archive is not obligate. Only stressing on public records is like what dictatorial government acted in past years. It is what we ignore diversity and request of community. We need to move our view that we have focused on public and central sectors to private and local sectors. Local records management based on locality could help to complete the entire puzzle. The way complete the puzzle is various and wide spheres including from cultural space to being extinct village. Locality is defined as the property in certain area or distinctiveness of locals. Establishing production strategies is as important as collecting records produced over the past years for local archiving. Local archiving has to be regionally conducted in phase. Moreover common wealth and recognition of communities are reflected in the acquisition process. In next to archiving local organizations and private records according to collection policy, methodology on local archiving needs for archive management and use in various public and private fields. This methodology could be possible by building a local archive networking tool. It is true that Local archiving is not familiar and clear yet. If we can turn the effort for public records we have made to endeavor for private sectors, we might expect big fruits in private sectors. We easily emphasis on globalization or internationalization, our daily lives start on our villages. Setting aside our small communities, such a puzzle of the whole would never be completed. This is good time to begin finding lost puzzle for future. The key that can find lost puzzles be held in archiving localities.

A study on the change effect of emission regulation mode on vehicle emission gas (배기가스 규제 모드 변화가 차량 배기가스에 미치는 영향 연구)

  • Lee, Min-Ho;Kim, Ki-Ho;Lee, Joung-Min
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.4
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    • pp.1108-1119
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    • 2018
  • As the interest on the air pollution is gradually rising at home and abroad, automotive and fuel researchers have been studied on the exhaust and greenhouse gas emission reduction from vehicles through a lot of approaches, which consist of new engine design, innovative after-treatment systems, using clean (eco-friendly alternative) fuels and fuel quality improvement. This research has brought forward two main issues : exhaust emissions (regulated and non-regulated emissions, PM particle matter) and greenhouse gases of vehicle. Exhaust emissions and greenhouse gases of automotive had many problem such as the cause of ambient pollution, health effects. In order to reduce these emissions, many countries are regulating new exhaust gas test modes. Worldwide harmonized light-duty vehicle test procedure (WLTP) for emission certification has been developed in WP.29 forum in UNECE since 2007. This test procedure was applied to domestic light duty diesel vehicles at the same time as Europe. The air pollutant emissions from light-duty vehicles are regulated by the weight per distance, which the driving cycles can affect the results. Exhaust emissions of vehicle varies substantially based on climate conditions, and driving habits. Extreme outside temperatures tend to increasing the emissions, because more fuel must be used to heat or cool the cabin. Also, high driving speeds increases the emissions because of the energy required to overcome increased drag. Compared with gradual vehicle acceleration, rapid vehicle acceleration increases the emissions. Additional devices (air-conditioner and heater) and road inclines also increases the emissions. In this study, three light-duty vehicles were tested with WLTP, NEDC, and FTP-75, which are used to regulate the emissions of light-duty vehicles, and how much emissions can be affected by different driving cycles. The emissions gas have not shown statistically meaningful difference. The maximum emission gas have been found in low speed phase of WLTP which is mainly caused by cooled engine conditions. The amount of emission gas in cooled engine condition is much different as test vehicles. It means different technical solution requires in this aspect to cope with WLTP driving cycle.