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Studies on the Survival Rates after Ultrarapidly Frozen-Thawing of Porcine Embryos (돼지 수정난의 초급속 동결 융해후의 생존성에 관한 연구)

  • 김상근;이봉구
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.125-131
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    • 1992
  • This study was carried out to investigate the effects of concentration and equilibration time of cryoprotective agents on the survival rate of slowly and ultrarapidly frozen porcine embryos. The porcine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 38$^{\circ}C$ water bath. Survival rate was defined as development rat to the morula and blastocyst stage after in vitro culture or by FDA test. The results are summarized as follows : 1. The survival rates of porcine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0, 2.5, 3.0, 3.5, or 4.0M glycerol was 65.3, 61.8, 64.3, 59.4 or 39.4%, respectively. Addition of 0.25M sucrose into the freezing medium containing 2.0M glycerol showed higher survival rate than those of 2.5~4.0M glycerol. 2. The survival rates of porcine embryos after ultraradpid frozen-thawing in the freezing medium of 0.25M sucroese added 2.0, 2.5, 3.0, 3.5 or 4.0M DMSO was 65.6, 67.6, 68.6, 60.6 or 23.6%, respectively. However, addition of 0.25M sucrose into the freezing medium containing 3.0M DMSO showed higher survival rate than those of 2.0, 2.5, 3.5 or 4.0M DMSO. 3. The survival rates of porcine embryos after ultrapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0, 2.5, 3.0, 3.5 or 4.0M propanediol was 63.2, 60.3, 62.1, 52.3 or 24.3%, respectively. Addition of 0.25M sucroese into the freezing medium containing 2.0M propanediol showed higher survival rate than those of 2.5~4.0M glycerol. 4. The survival rates of porcine embryos after ultrarapid frozen-thawing the freezing medium of 2.0M glycerol added 0.10, 0.25, 0.50 or 0.75M sucrose was 61.8, 70.8, 67.6 or 52.2%, respectively. Addition of 2.0M glycerol into the freezing medium containing 0.25M sucreose showed higher survival rate than that those of 0.10, 0.50 or 0.75M sucrose. 5. The higher suvival rate of porcine embryos were attained at short period of equilibration time 92.5~5min.) in the freezing medium added 0.25M sucreose and 3.0M compared to those of 10 or 20min. equilibration time in the same condition.

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A Study on the Influence Area of Excavation around Railroads (철로 주변의 지하굴착 영향권에 관한 연구)

  • Park, Jong-Su;Jang, Jeong-Wook;Park, Choon-Sik
    • Proceedings of the Korean Geotechical Society Conference
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    • 2006.03a
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    • pp.1032-1037
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    • 2006
  • This thesis studied effects of the excavation around railroads on the deformation of the lateral ground and neighboring railroads. The conclusions of the study are as follows. 1. When the depth of excavationis 10m, the influential area should be 35m for soft clay, 20m for normal clay, 15m for hard clay, 15m for loose sand, 12m for slightly dense sand, and 8m for dense sand. 2. When the influential area is 10m, the allowable excavation depth should be 2.5m for soft clay, 4.8m for normal clay, 7.5m for hard clay, 7.2m for loose sand, 8.8m for slightly dense sand, and 10m for dense sand. 3. When the influential area is 20m, the allowable excavation depth should be 4.5m for soft clay, and up to 10m for the other five kinds of soil. 4. When the influential area is 30m, the allowable excavation depth should be 7.5m for soft clay, and up to 10m for the other five kinds of soil. 5. When the influential area is 35m, the allowable excavation depth should be up to 10m for all kinds of soil.

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Study on the Enzyme of Basidiomycetes(I) -The Effects of Iron Ions on the Light-Induced Mitochondrial $F_0F_1-ATPase$ of Lentinus edodes- (담자균류의 효소에 관한 연구(I) -표고버섯 중의 광감응성 Mitochondrial $F_0F_1-ATPase$의 철이온 효과-)

  • Min, Tae-jin;Lee, Mi-Ae;Bae, Kang-Gyu
    • The Korean Journal of Mycology
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    • v.21 no.3
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    • pp.165-171
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    • 1993
  • The effects of the iron ions for the light-induced mitochondrial $F_0F_1-ATPase$ of Lentinus edodes was studied. The enzyme activity was stimulated up to 202% by 0.1 mM $Fe^{2-}$ ion, but was inhibited by $Fe^{3+}\;and\;Mg^{2+}$. In the presence of 0.5 mM $Mg^{2+}$, the activity also increased 32% by 0.1 mM $Fe^{2+}$ ion, and decreased to a similar extent by $Fe^{3+}$ ion than by only $Fe^{3+}$ ion. Also, the activity was inhibited 53% by 5.0 mM $Fe^{2-}$ ion in the presence of 0.5 mM $Mg^{2+}$ ion and various concentration of $Fe^{3+}$ ion(mM). These results showed that $Fe^{2+}$ strongly stimulated the enzyme activity and its role for the enzyme was independent of $Mg^{2+}$ ion, but was dependent of $Fe^{3+}$ ion. From inactivation of the enzyme by addition of metal chelating agent, EDTA, it is suggested that the enzyme is to be metalloenzyme. The optimal pH and temperature of the enzyme in the presence of 0.1 mM $Fe^{2+}$ was 7.6 and $63^{\circ}C$, respectively.

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Effects of High Glucose on Na,K-ATPase and Na/glucose Cotransporter Activity in Primary Rabbit Kidney Proximal Tubule Cells

  • Han, Ho-Jae
    • The Korean Journal of Physiology
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    • v.29 no.1
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    • pp.69-80
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    • 1995
  • Renal proximal tubular hypertrophy and hyperfunction are known to be early manifestations of experimental and human diabetes. As the hypertrophy and hyperfunction have been suggested to be central components in the progression to renal failure, an understanding of their underlying causes is potentially important for the development of therapy. A primary rabbit kidney proximal tubule cell culture system was utilized to evaluate the possibility that the renal proximal tubular hypertrophy and hyperfunction observed in vivo in diabetes mellitus, can be attributed to effects of elevated glucose levels on membrane transport systems. Primary cultures of rabbit proximal tubules, which achieved confluence at 10 days, exhibited brush-border characteristics typical of proximal tubular cells. Northern analysis indicated $2.2{\sim}2.3$ and 2.0 kb Na/glucose cotransporter RNA species appeared in fresh and cultured proximal tubule cells after confluence, repectively. The cultured cells showed reduced Na/glucose cotransporter activity compared to fresh proximal tubules. Primary cultured proximal tubule cells incubated in medium containing 20 mM glucose have reduced ${\alpha}-MG$ transport compared to cells grown in 5 mM glucose. In the proximal tubule cultures incubated in medium containing 5 mM or 20 mM glucose, phlorizin at 0.5 mM inhibited 0.5 mM ${\alpha}-MG$ uptake by 84.35% or 91.85%, respectively. The uptake of 0.5 mM ${\alpha}-MG$ was similarly inhibited by 0.1 mM ouabain (41.97% or 48.03% inhibition was observed, respectively). In addition, ${\alpha}-MG$ uptake was inhibited to a greater extent when $Na^{+}$ was omitted from the uptake buffer (81.86% or 86.73% inhibition was observed, respectively). In cell homogenates derived from the primary cells grown in 5 mM glucose medium, the specific activity of the Na/K-ATPase $(6.17{\pm}1.27\;{\mu}mole\;Pi/mg\;protein/hr)$ was 1.56 fold lower than the values in cell homogenates treated with 360 mg/dl D-glucose, 20 mM $(9.67{\pm}1.22\;{\mu}mole\;Pi/mg\;protein/hr)$. Total $Rb^{+}$ uptake occurred at a significantly higher rate (1.60 fold increase) in primary cultured rabbit kidney proximal tubule cell monolayers incubated in 20 mM glucose medium $(10.48{\pm}2.45\;nM/mg\;protein/min)$ as compared with parallel cultures in 5 mM glucose medium. $Rb^{+}$ uptake rate in 5 mM glucose medium was reduced by 28% when the cultures were incubated with 1 mM ouabain. The increase of the $Rb^{+}$ uptake by rabbit kidney proximal tubule cells in 20 mM glucose could be attributed primarily to an increase in the rate of ouabain-sensitive $Rb^{+}$ uptake $(5\;mM\;to\;20\;mM;\;4.68{\pm}0.85\;to\;8.38{\pm}1.37\;nM/mg\;protein/min)$. In conclusion, the activity of the renal proximal tubular Na,K-ATPase is elevated in high glucose concentration. In contrast, the activity of the Nafglucose cotransport system is inhibited.

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Fruit Quality Characteristics of 'Fuji' Apple Fruits in Response to Air Storage after CA Storage (Controlled Atmosphere (CA) 환경설정 해제 후 저온저장이 '후지' 과실의 품질에 미치는 영향)

  • Kweon, Hun-Joong;Choi, Dong Geun;Lee, Jinwook;Kang, In-Kyu
    • Horticultural Science & Technology
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    • v.32 no.2
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    • pp.178-183
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    • 2014
  • This study aimed to evaluate the fruit quality characteristics and incidence of flesh browning in response to air storage at $0{\pm}1^{\circ}C$ a fter controlled atmosphere s torage (CA condition: $O_2$ $2.5{\pm}0.5$%, $CO_2$ $1.5{\pm}0.5$%) at $0{\pm}1^{\circ}C$ in 'Fuji' apples (Malus domestica Borkh.). The storage system was performed as followed: air storage for one month, CA storage 4 months + air storage 3 months ( CA 4M + A ir 3M), CA storage 5 m onths + air storage 2 months ( CA 5M + Air 2M) a nd C A storage 6 m onths + air storage 1months (CA 6M + Air 1M), while the control fruits were stored at CA storage for 8 months right after harvest. The incidence of flesh browning ranged from 17.1% to 30.2% during CA storage but not detected under the treatments of CA 4M + Air 3M and CA 5M + Air 2M. The respiration rate was not affected by storage treatments for 6M while the respiration rate was lower in the treatments of CA 4M + Air 3M and CA 5M + Air 2M than the other storage treatments after 7 months. Ethylene production and internal ethylene concentration were lowest in rapid CA storage and increased with a decreasing CA storage duration. Therefore, the results indicate that CA 5M + Air 2M storage treatment should be recommended to maintain the fruit quality and reduce the risk development of flesh browning rather than typical CA storage in 'Fuji' apples.

Bacillus stearothermophilus 에서 부분 정제한 Cytosine Deaminase 의 특성

  • 장영채;이경형;김성영;조윤래;김종규
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.305-309
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    • 1992
  • Cytosine deaminase (EC 3.5.4.1) from BaciNus stc~urorhermophilus was partially purified 7.2-fold with an overall yield of 52.7%. The partially purified enzyme deiiminated cytosine only.but not 5-methylcytosine and 5-fluorocytosine. The apparent Michaclis constant. Km valuefor cytosine was 5.9 mM. The enzyme was relatively stable in the range of pH 4.0 to 7.0.furthermore extremely thermo-stable : more than 75'X) of the activity was remained afterheating at 80$^{\circ}$C for I0 min at pH 6.5. The enzyme had a pH optimum at around pH7.0 to 7.5. and temperature optimum at 35 to 31$^{\circ}$C. And the activation energ (En value)determined from an Arrhenius plot was 26 Kcal/mol. The enzyme activity was stronglyinhibited by heavy metal ions such as Cd", Hg". Cut' at 1 mM, anJ by o-phenanthroline,and p-chloromcrcuribcnzoate at I mM. But the enrymc activity was activatetl increased byGMP, and CMP at 1 mM.ased by GMP, and CMP at 1 mM.

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Purification and Properties of Extracellular Cytosine Deaminase from Chromobacterium violaceum YK 391

  • Yu, Tae-Shick;Kim, Tae-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.173-178
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    • 1999
  • The extracellular cytosine deaminase (EC 3.5.4.1) from Chromobacterium violaceum YK 391 was purified 264.7-fold with an overall yield of 14.3%. The enzyme was for the first time homogeneous by the criteria of polyacrylamide gel electrophoresis performed in the absence and in the presence of sodium dodecyl sulfate. The molecular weight of the purified enzyme was estimated to be about 156 kDa. The enzyme consisted of two identical subunits of approximate molecular weight 78 kDa. The isoelectric point of the enzyme was pH 5.55. The enzyme had a pH optimum of 7.5 and a temperature optimum of around 40 to $45^{\circ}C$. Besides cytosine, the enzyme deaminated 5-fluorocytosine, cytidine, 5-methylcytosine, and 6-azacytosine, but not 5-azacytosine. The extracellular cytosine deaminase is believed to be unique because it was active not only on cytosine but also on cytidine. The apparent $K_m$ values for cytosine, 5-fluorocytosine, cytidine, and 5-methylcytosine were determined to be 1.55 mM, 5.52 mM, 10.4 mM, and 67.2 mM, respectively. The enzyme activity was strongly inhibited by heavy metal ions such as $Fe^{2+},Pb^{2+},Cd^{2+},Zn^{2+}, Hg^{2+}, and Cu^{2+}$ at 1 mM, and completely by $\alpha,\alpha$'-dipyridyl, and $\rho$-chloromercuribenzoate at 1 mM, and weakly inhibited by 1mM ο-phenanthroline. The enzyme activity was not affected by various nucleosides and nucleotides.

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The Evaluation of Safe Yield Considered Interference Drawdown between Hot Spring Wells at Yecheon Hot Spring Area (예천온천지구내 온천공간 수위간섭을 고려한 적정양수량 평가)

  • Lee, Chol-Woo
    • The Journal of Engineering Geology
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    • v.17 no.3
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    • pp.405-409
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    • 2007
  • There exist 5 wells drilled at the Yecheon hot spring area, the distance between the wells is about $159m{\sim}702.6m$. The safe yield of each well is controlled by interference drawdown between a pumping well and an observation well after pumping test. The well No. 2 and the well No. 5 are the closest, at the distance of 159 m; therefore interference drawdown between two wells was occurred considerably. The drawdown of the well No. 2 (an observation well) was 16.67 m; the drawdown of the well No. 5 (a pumping well) was 17.21 m. The degree of the interference is about 97% and the safe yield decreased from $750m^3/day\;to\;24m^3/day$. Significant interference between two wells is due to the fact that the aquifer has 1.5 dimensions.

Quality and Fruit Productivity of the Second Truss Blooming Seedlings Depending on Concentration of Nutrient Solution in Cherry Tomato (양액 농도에 따른 방울토마토 2화방 개화묘의 소질 및 과실 생산성)

  • Lee, Mun Haeng
    • Journal of Bio-Environment Control
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    • v.31 no.3
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    • pp.230-236
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    • 2022
  • This study was carried out to produce two-flowered seedlings, harvest them early in a greenhouse, and extend the harvest period. This study was carried out to effectively produce the second truss blooming seedlings to harvest tomatoes early and extend the harvest period. For production of the second truss blooming seedlings (one stem), the nutrient solution EC was supplied at 1.5, 2.0, 2.5 dS·m-1, and dynamic management (3.0 → 3.5 → 4.5 dS·m-1). The seedling period was 60 days, which was 20-40 days longer than conventional seedlings, and 10 days longer than the first truss blooming seedlings (cube seedlings). The plant height was 78 and 77 cm in EC 2.5 dS·m-1 and dynamic management respectively, which was shorter than EC 1.5 dS·m-1 with 88 cm. As for the EC in the cube before formulation, dynamic management had the highest EC 5.5 dS·m-1, and the cube supplied with EC 1.5 dS·m-1 had the lowest. The production yield by treatment did not a difference among in the second truss blooming seedlings, but the first truss blooming seedlings showed lower productivity than second truss blooming seedlings. The second truss blooming seedling were harvested 35 days after planting on June 4, the first harvest date, and the first truss blooming were harvested in 42 days on June 11th. There was no difference in plant height and root growth due to bending at frequency planting. In the study on the production of the second truss blooming seedlings (two stem), the nutrient solution EC was supplied under 2.0, 2.5, 3.0 dS·m-1, and dynamic management (3.0 → 3.5 → 4.5 dS·m-1). The seedling period was 90 days, which was 40-50 days longer than conventional seedlings and 10 days longer than the first truss blooming seedlings (cube seedlings). Plant height was 80 and 81 cm in EC 2.0 dS·m-1 and 2.5 dS·m-1 respectively, but was the shortest at 73 cm in dynamic management. EC in the medium increased as the seeding period increased in all treatments. The dynamic management was the highest with EC 5.1 dS·m-1. There was no difference in yield among EC treatments in the second truss blooming seedlings, which had a longer seeding period of about 10 days, produced 15% more than the first truss blooming seedlings. In order to shorten the plant height of the second truss blooming seedlings, it is judged that the most efficient method is increasing the concentration of nutrient solution.

The Combination Effect of Kinetin and Auxin on the Growth Root Development and Vitamin C Content of Soybean sprouts (콩나물의 성장과 Vitamin C 생성에 미치는 Kinetin과 Auxin의 혼합효과)

  • Kim, Sang-Ock
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.11 no.2
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    • pp.37-41
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    • 1982
  • This study was conducted to investigate the growth of soybean sprouts and the effective hormonal refutation of vitamin C content by the concentration and combination of kinetin and auxins such as IAA, 2.4-D and NAA. The results were as follows; 1. The growth of soybean sprouts were fine under the respective conditions of kinetin $10^{-8}M+IAA\;10^{-6}M$, kinetin $10^{-7}M+IAA\;10^{-5}M$, kinetin $10^{-6}M+IAA\;10^{-6}M$, kinetin $10^{-5}M+IAA\;10^{-7}M$, kinetin $10^{-8}M+2,4-D\;10^{-8}M$ and kinetin $10^{-7}M+NAA\;10^{-5}M$. 2. The root development of soybean sprouts were almost in accord with the growth of soybean sprouts by the concentration of kinetin and auxin. 3. The content of vitamin C were more increased under in kinetin $10^{-8}M+IAA\;10^{-8}M$, kinetin $10^{-6}M+2,4-D\;10^{-5}M$ and kinetin $10^{-8}M+NAA\;10^{-8}M$ and both growth and vitamin C content were more activated under kinetin $10^{-8}M+IAA\;10^{-6}M$ and $10^{-8}M+NAA\;10^{-8}M$. 4. The growth, root development and vitamin C content were increased in the low kinetin and high IAA, NAA concentration. Root development and vitamin C content of soybean hypocotyl were decreased in the low kinetin and high 2,4-D concentration.

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