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Studies on Protoplast Formation and Regeneration of Coriolus versicolor (구름버섯의 원형질체(原形質體) 형성(形成)과 재생(再生)에 관한 연구(硏究))

  • Bok, Jin-Woo;Park, Seol-Hee;Choi, Eung-Chil;Kim, Byong-Kak;Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.18 no.3
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    • pp.115-126
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    • 1990
  • To establish basic techniques for protoplast fusion of Coriolus versicolor several factors affecting protoplast formation and regeneration were investigated. Protoplast isolation was at maximum with 2.5-day cultured mycelia of C. versicolor treated with the combination of two enzymes, Novozym 234 (10 mg/ml) and cellulase Onozuka R-10 (15 mg/ml), for 3-4.5 hours at $30^{\circ}C.$ As an osmotic stabilizer for stabilizing the protoplast, 0.6 M sucrose was the best for formation and regeneration of the protoplast from the mycelia of the fungus and the regeneration frequency was 3.48%. Protoplast fusion was made by a modified method of Peberdy using PEG (M.W. 4,000). The fusion frequency between two mutants of C. versicolor was 1.86% and the fusion products showed differences in growth rate and colony morphology.

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Strain Improvement of Aspergillus oryzae for Increasing Productivity of a Proteolytic Enzyme. (고활성 단백질분해효소 생산균주의 개발을 위한 Aspergillus oryzae의 원형질체 융합에 의한 변이)

  • 김두상;김형락;남택정;변재형
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.490-496
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    • 1998
  • Aspergillus oryzae producing high proteolytic enzyme was isolated from soybean koji and named tentatively A. oryzae O-1. A. oryzae U-1 was obtained by mutation of A. oryzae O-1 with ultraviolet (UV)-irradiation and produced 14 times higher pretense activity compared with A. oryzae O-1. A. oryzae E-1 was acquired by treatment of A. oryzae U-1 with 0.5 M ethylmethanesulfonate (EMS) for 6 min at 3$0^{\circ}C$ and produced 39 times higher proteolytic activity than A. oryzae O-1. With protoplast fusion between A. oryzae O-1 and A. oryzee E-1 in the presence of polyethylenegylcol (PEG)-CaCl$_2$, proteolytic activity was increased to 82 times compared to A. oryzae O-1, and the fusant was named A. oryzae PF. The activities of the cultures containing proteolytic enzymes produced by the strains were determined to be 0.23 U/$m\ell$ for A. oryzae O-1, 3.29 U/$m\ell$ for A. oryzae U-1, 8.91 U/$m\ell$ for A. oryzae E-1, and 19.0 U/$m\ell$ for A. oryzae PF.

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The Consideration for Optimum 3D Seismic Processing Procedures in Block II, Northern Part of South Yellow Sea Basin (대륙붕 2광구 서해분지 북부지역의 3D전산처리 최적화 방안시 고려점)

  • Ko, Seung-Won;Shin, Kook-Sun;Jung, Hyun-Young
    • The Korean Journal of Petroleum Geology
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    • v.11 no.1 s.12
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    • pp.9-17
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    • 2005
  • In the main target area of the block II, Targe-scale faults occur below the unconformity developed around 1 km in depth. The contrast of seismic velocity around the unconformity is generally so large that the strong multiples and the radical velocity variation would deteriorate the quality of migrated section due to serious distortion. More than 15 kinds of data processing techniques have been applied to improve the image resolution for the structures farmed from this active crustal activity. The bad and noisy traces were edited on the common shot gathers in the first step to get rid of acquisition problems which could take place from unfavorable conditions such as climatic change during data acquisition. Correction of amplitude attenuation caused from spherical divergence and inelastic attenuation has been also applied. Mild F/K filter was used to attenuate coherent noise such as guided waves and side scatters. Predictive deconvolution has been applied before stacking to remove peg-leg multiples and water reverberations. The velocity analysis process was conducted at every 2 km interval to analyze migration velocity, and it was iterated to get the high fidelity image. The strum noise caused from streamer was completely removed by applying predictive deconvolution in time space and ${\tau}-P$ domain. Residual multiples caused from thin layer or water bottom were eliminated through parabolic radon transform demultiple process. The migration using curved ray Kirchhoff-style algorithm has been applied to stack data. The velocity obtained after several iteration approach for MVA (migration velocity analysis) was used instead or DMO for the migration velocity. Using various testing methods, optimum seismic processing parameter can be obtained for structural and stratigraphic interpretation in the Block II, Yellow Sea Basin.

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Antioxidant Activity and Physiological Function of the Anomala albopilosa Extracts (청동풍탱이(Anomala albopilosa)추출물의 항산화성 및 생리기능)

  • Yoon, Weon-Jong;Lee, Jung-A;Kim, Ji-Young;Kim, Sang-Bum;Park, Soo-Yeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.6
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    • pp.670-677
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    • 2007
  • We analyzed antioxidant and physiological activities to investigate the functional effects of ethanol extracts of Anomala albopilosa imago and A. albopilosa larva. In order to effectively screen for anti-inflammatory agents, we first investigated the antioxidant activities such as DPPH radical scavenging capacity, superoxide radical scavenging capacity, xanthine oxidase inhibitory activity, and nitric oxide scavenging capacity of the A. albopilosa extracts. By the screening system, we found that A. albopilosa extracts had antioxidant activity which increased with increments of the extract concentration. Moreover, we examined the inhibitory effect of the A. albopilosa extracts on the production of anti-inflammatory factors that the nitric oxide (NO), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and prostaglandin $E_2\;(PGE_2)$ production activated with LPS ($1{\mu}g/mL$) in murine macrophage cell line RAW 264.7. A. albopilosa extracts potentially inhibited the iNOS and COX-2 in a dose-dependent manner. The inhibition of iNOS activity was correlated with the decrease in nitrite levels. Additionally, the $PEG_2$ production is markedly inhibited after a treatment with the A. albopilosa extracts.

Evaluation of Vascularization of Hydroxyapatite Ocular Implants by $^{99m}Tc-MDP$ Bone Scan : Preliminary Study ($^{99m}Tc-MDP$ 골주사를 이용한 Hydroxyapatite 안구보충물의 혈관신생에 관한 예비연구)

  • Kim, Jong-Deok;Shin, Sung-Gyun;Lee, Sung-Woo
    • The Korean Journal of Nuclear Medicine
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    • v.29 no.4
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    • pp.518-525
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    • 1995
  • 목적 : 안구제거술 후에 삽입한 hydroxyapatite 안구보충물내로 혈관신생이 완전하게 일어 나는 시기를 알아보기 위함이다. 방법 : 안구제거술을 받은 43예의 환자(남자 33예, 여자 10예, 14-56세)에게 대부분 변경되지 않은 방법으로 hydroxyapatite 안구보충물을 삽입한 후 10-23주 사이에 다양한 간격(10-12 주 2예, 13-16주 18예, 17-20주 20예, 21-23주 3예)을 두고 $^{99m}Tc-MDP$ 골주사를 시행하여 보충물내로의 혈관신생 정도를 전향적으로 측정하였다. 정면 및 측면촬영의 정적영상에서 안구보충물의 방사능섭취가 비교(nasal bridge)의 방사능섭취보다 약할 경우를 grade 1, 같을 경을 grade 2, 강할 경우를 grade 3로 나누었으며 grade 2와 3을 혈관신생이 완전하다고 정의하였다. 또한 43예 모두에서 방사능섭취 정도와 관계없이 안구보충물 삽입후 평균 21주되는 시기에 안구보충물에 구멍을 내어 이 구멍으로부터의 출혈여부를 확인한 후 운동성나사(motility peg)를 끼워 의안(prosthesis)과 연결하였다. 43예중 7예에서는 $^{99m}Tc-MDP$ 골주사와 조영제주입후의 자기공명영상(Gd-DTPA T1-weighted images)을 같은 날짜에 시행하여 각각의 예에서 방사능섭취 정도와 조영제증강 정도를 비교하였다. 결과 : 안구보충물내로 완전하게 혈관신생이 일어났던 grade 2와 3의 비율은 안구보충물삽입후 10-12주에서는 0%, 13-16주 33%, 17-20주 50%, 21-23주 67%로써 시간경과에 따라 점차 증가하였다. 골주사소견에 관계없이 안구보충물 삽입후 평균 21주되는 시기에 보충물 앞쪽에 구멍을 내었을 때 43예 모두에서 완전한 혈관신생을 의미하는 출혈을 볼 수 있었다. $^{99m}Tc-MDP$ 골주사와 조영제주입후의 자기공명영상을 둘 다 시행한 7예 각각에서 방사능섭취의 정도와 조영제증강의 정도가 거의 일치하였다. 결론 : 본 예비연구의 결과로써 hydroryapatite 안구보충물 삽입후 21주째에 $^{99m}Tc-MDP$ 골주사를 시행하는 것이 좋을 것으로 생각되며, hydroxyapatite 안구보충물내로의 완전한 혈관신생 여부를 보는 데에는 $^{99m}Tc-MDP$ 골주사가 조영제주입후의 자기공명영상보다 더 경제적이라고 할 수 있겠다.

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Protoplast Formation and Fusion between Saccharomyces cerevisiae D-71 and Zygosaccharomyces rouxii SR-S (Saccharomyces cerevisiae D-71과 Zygosaccharomyces rouxii SR-S의 원형질체 형성과 융합)

  • 이종수;김찬조
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.142-149
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    • 1988
  • This experiment was carried out to obtain a hybrid with potent ethanol fermenting ability, by means of protoplast fusion between a thermophilic strain (D-71) of Saccharomyces cerevisiae and an osmotolerant strain (SR-S) of Zygosaccharomyces rouxii. The conditions for formation of protoplasts from both strains and for their fusion and regeneration were studied. Favorable conditions for formation of protoplasts from Saccharomyces cerevisiae D-71 were : treatment of the cells at late-exponential phase with 2-mercaptoethanol (l% v/v) for 10 minutes in the presence of 0.5M sorbitol, then incubation for 60 minutes in the set medium containing Zymolyase-20T (4mg/$m\ell$) ; and from Zygosaccharomyces rouxii SR-S were : treatment of the cells at mid-exponential phase with 2-mercaptoethanol (1% v/v) for 10 minutes in the presence of 0.5M or 1M mannitol, then incubation for 120 minutes in the set medium containing Zymolyase-20T(4mg/$m\ell$). The protoplasts of parental cells were fused in the presence of 20mM CaCl$_2$, 0.5M sorbitol and 40% of polyethyleneglycol (M.W 4000), then fusants obtained were selected as regenerated colonies which embedded and grown in the minimal medium containing 3% of agar. The frequencies of fusant formation were 1.2$\times$10$^{-6}$ to 9.1$\times$10$^{-6}$ for the regenerated protoplast.

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Induction of Somatic Hybrid by Protoplast Fusion between Populus koreana × P. nigra var. italica and P. euramericana cv. Guardi (수원포플러와 구아디 포플러 원형질체(原形質體) 융합(融合)에 의한 체세포잡종체(體細胞雜種體) 유도(誘導))

  • Park, Young Goo;Kim, Jung Hee;Son, Sung Ho
    • Journal of Korean Society of Forest Science
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    • v.81 no.3
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    • pp.273-279
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    • 1992
  • Protoplasts isolated from leaf mesophyll tissues of Populus koreana ${\times}$ P. nigra var. italica were fused with those of P. euramericana cv. Guardi. Well expended healthy leaves of 5 to 7 week-old-plantlet grown in vitro were used as source materials. Leaves from P. koreana ${\times}$ P. nigra var. italica and P. euramericana cv. Guardi were digested in enzyme solution I (2.0% Cellulase, 1.2% Hemicellulase, 0.4% Macrozyme, 2.0% Driselase, 0.05% Pectolyase ; w/v) and enzyme solution II (1.0% Cellulase, 1.2% Hemicellulase, 0.4% Macrozyme, 2.0% Driselase, 0.05% Pectolyase ; w/v), respectively, The highest frequency of fusion among the protoplasts originated from the two source materials was approximately 21% using 40% PEG or 15% dextran. In addition, fusion frequency was enhanced by incorporating 30mM of $Ca^{2+}$ in eluting solution at pH 10.5. Dividing cells and/or mint-calli were obtained by culturing the fusion products in a liquid 8p-KM medium supplemented with 0.6M sucrose, $0.45{\mu}M$ 2, 4-D, and $0.5{\mu}M$ BA. Shoots were regenerated from the fusion product-derived calli after culture on MS medium containing $5.0{\mu}M$ zeatin. To verify the putative hybrid or cybrid, SDS-PAGE was carried out. From the 24 regenerants, just two plants showed intermediate protein band patterns compared with those of the original source plants.

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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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Study of the Sludge Formation Mechanism in Advanced Packaging Process and Prevention Method for the Sludge (어드밴스드 패키징 공정에서 발생할 수 있는 슬러지의 인자 확인 및 형성 방지법의 제안)

  • Jiwon Kim;Suk Jekal;Ha-Yeong Kim;Min Sang Kim;Dong Hyun Kim;Chan-Gyo Kim;Yeon-Ryong Chu;Neunghi Lee;Chang-Min Yoon
    • Journal of the Korea Organic Resources Recycling Association
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    • v.31 no.1
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    • pp.35-45
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    • 2023
  • In this study, the sludge formation in the wastewater drain from the advanced packaging process mechanisms are revealed as well as the key factors, materials, and sludge prevention methods using surfactant. Compared with that of conventional packaging process, advanced packaging process employ similar process to the semiconductor fabrication process, and thus many processes may generate wastewater. In specific, a large amount of wastewater may generate during the carrier wafer bonding, photo, development, and carrier wafer debonding processes. In order to identify the key factors for the formation of sludge during the advanced packaging process, six types of chemicals including bonding glue, HMDS, photoresist (PR), PR developer, debonding cleaner, and water are utilized and mixing evaluation is assessed. As a result, it is confirmed that the black solid sludge is formed, which is originated by the sludge seed formation by hydrolysis/dehydration reaction of HMDS and sludge growth via hydrophobic-hydrophobic binding with sludge seed and PR. For the sludge prevention investigation, three surfactants of CTAB, PEG, and shampoo are mixed with the key materials of sludge, and it is confirmed that the sludge formations are successfully suppressed. The underlying mechanism behind the sludge formation is that the carbon tails of the surfactant bind to PR with hydrophobic-hydrophobic interaction and inhibit the reaction with HMDS-based slurry seeds to prevent the sludge formation. In this regard, it is expected that various problems like clogging in drains and pipes during the advanced packaging process may effectively solve by the injection of surfactants into the drains.

Efficient Selection Method for Drought Tolerant Plants Using Osmotic Agents

  • Park, Dong-Jin;Im, Hyeon-Jeong;Jeong, Mi-Jin;Song, Hyeon-Jin;Kim, Hak-Gon;Suh, Gang-Uk;Ghimire, Balkrishna;Choi, Myung-Suk
    • Journal of Forest and Environmental Science
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    • v.34 no.3
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    • pp.224-234
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    • 2018
  • An efficient method to select drought tolerant Korean native plants using in vitro culture system was established in this study. While the plant growths and root inductions of each plant were proportionately affected by concentrations of mannitol on in vitro culturing seven plant species to test tolerance to osmotic stress, growth index (GI) and number of root induction of Chrysanthemi zawadskii var. latilobum and Dianthus chinensis var. semperflorens plantlets were higher than the others in 125mM mannitol. In test with polyethylene glycol (PEG), plantlets of C. zawadskii var. latilobum and D. chinensis var. semperflorens showed higher GI and number of root induction than the others in 33.3mM. On testing whether the well grown plants under osmotic stress are tolerant to virtual drought stress, there were significant differences in the withering rates of C. zawadskii var. latilobum and D. chinensis and those of were Aster yomena and Centaurea cyanus after 12 days without watering. It was found that significantly lower stomata numbers were shown in both drought tolerant plants than the sensitive plants. Averages of the stomata circumferences and the stomata area in the plantlets of the tolerant species were larger than those of the sensitive plants D. chinensis var. semperflorens showed the lowest transpiration level per unit area. The highest stomatal area per unit area was found in C. zawadskii, followed by D. chinensis var. semperflorens, Aster yomena and C. cyanus. In conclusion, C. zawadskii var. latilobum and D. chinensis var. semperflorens were more tolerant to drought than other two species. Furthermore in vitro selection was successfully used to screen drought tolerance species of native plant species.