• Title/Summary/Keyword: culture conditions

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The Effect of Platelets on Endothelin Production in Bovine Pulmonary Artery Endothelial Cells (혈소판이 소 폐동백 내피세포의 Endothelin 생산에 미치는 효과)

  • Lee, Sang-Do;Shim, Tae-Sun;Kwon, Seog-Woon;Ryu, Jin-Sook;Lee, Jae-Dam;Lim, Chae-Man;Koh, Youn-Suck;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.5
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    • pp.1114-1124
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    • 1997
  • Background : Endothelin(ET) is a very potent vasoconstrictive peptide produced by endothelial cells of pulmonary artery. The endothelin level was increased in plasma of primary pulmonary hypertension and acute pulmonary thromboembolism and it was suggested that the endothelin might do a critical role in the cardiopulmonary dysfunction in these two conditions. But the exact mechanism of increase of ET has not been known. In these two conditions, platelet activation and thrombosis are the main pathophysiologic findings. So there is a possibility that the platelet might stimulate endothelin secretion from endothelial cells. Therefore, we performed this study to evaluate the role of platelet and its mediators on endothelin production in bovine pulmonary artery endothelial(BPAE) cells. Method : Bovine pulmonary artery endothelial cells, ATCC certified cell line 209, were cultured and treated with human platelets($10^6{\sim}10^8/ml$), thrombin (0.1~10u/ml), TGF-${\beta}1$(1~100uM), serotonin(1~100uM), and endotoxin(1ug/ml) in a final volume of 500ul for 18 hours. Levels of ir(immunoreactive)-ET in each conditioned medium were measured by a radioimmunoassay specific for ET. Result : The increase of ir-ET levels was platelet number and time dependent over 18 hours. When washed human platelets were added($10^8/ml$), the ir-ET levels were significantly higher than that of control(p<0.05) at 8 and 18 hours after culture. Subthreshold concentration of platelets($10^7/ml$) coincubated with endotoxin(1ug/ml) or subthreshold dose of thrombin(0.1u/ml) stimulated ir-ET secretion from BPAE cells significantly(p<0.05) compared with control. Thrombin(1ug/ml, 10ug/ml) and TGF-${\beta}1$(100pM, 1000pM) significantly increased ir-ET secretion from BP AE cells(p<0.05) compared with control, but serotoin(1~100uM) and endotoxin(1ug/ml) did not stimulate the ir-ET secretion. Conclusions : Platelets stimulate endothelin secretion from bovine pulmonary artery endothelial cells. The mechanism of increase of endothelin secretion seems to be a stimulation by platelet itself or by mediators, such as TGF-${\beta}1$, secreted from activated platelets. And, in this study, the priming effect of platelets on endothelin secretion from BPAE cells could be another possibility.

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Culture Conditions of Aspergillus oryzae in Dried Food-Waste and the Effects of Feeding the AO Ferments on Nutrients Availability in Chickens (건조한 남은 음식물을 이용한 Aspergillus oryzae균주 배양조건과 그 배양물 급여가 닭의 영양소 이용률에 미치는 영향)

  • Hwangbo J.;Hong E. C.;Lee B. S.;Bae H. D.;Kim W.;Nho W. G.;Kim J. H.;Kim I. H.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.291-300
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    • 2005
  • Two experiments were carried out to assess the appropriate incubation conditions namely; duration, moisture content and the ideal microbial inoculant for fermented dried food waste(EW) offered to broilers. The nutrient utilization of birds fed the FW diets at varying dietary inclusion rates was also compared with a control diet. In Experiment 1, different moisture contents(MC) of 30, 40, 50 and $60\%$ respectively were predetermined to establish the ideal duration of incubation and the microbial inoculant. A 1mL Aspergillus oryzae(AO) $(1.33\times10^5\;CFU/mL)$ was used as the seed inoculant in FW. This results indicated that the ideal MC for incubation was $40\~50\%$ while the normal incubation time was > 72 hours. Consequently, AO seeds at 0.25, 0.50, 0.75 and 1.00mL were inoculated in FW to determine its effect on AO count. The comparative AO count of FW incubated for 12 and 96 hours, respectively showed no significant differences among varying inoculant dosage rates. The FW inoculated with lower AO seeds at 0.10, 0.05 and 0.01mL were likewise incubated for 72 and 96 hours, respectively and no changes in AO count was detected(p<0.05). The above findings indicated that the incubation requirements for FW should be $%40\~50\%$ for 72 hours with an AO seed incoulant dosage rate of 0.10mL. Consequently, in Experiment II, after determining the appropriate processing condition for the FW, 20 five-week old male Hubbard strain were used in a digestibility experiment. The birds were divided into 4 groups with 5 pens(1 bird per pen). The dietary treatments were; Treatment 1 : Control(Basal diet), Treatment 2 : $60\%$ Basal+4$40\%$ FW, Treatment 3 : $60\%$ $Basal+20\%\;FW+20\%$ AFW(Aspergillus oryzae inoculate dried food-waste diet) and Treatment 4: $60\%$ Basal+$40\%$ Am. Digestibility of treatment 2 was lowed on common nutrients and amino acids compared with control(p<0.05) and on crude fat and phosphorus compared with AFW treatments(T3, T4)(plt;0.05). Digestibility of treatment 3 and 4 increased on crude fiber and crude ash compared treatment 2 (p<0.05). Digestibility of control was high on agrinine, leucine, and phenylalnine of essential amino acids compared with treatment 3 and 4(p<0.05), and diestibility of treatment 3 and 4 was improved on arginine, lysine, and threonine of essential amino acids. Finally, despite comparable nutrient utilization among treatments, birds fed the dietary treatment containing AO tended to superior nutrient digestion to those fed the $60\%$ Basa1+$40\%$ FW.

A Study of Improvement Plans for Urban Parks through the Application of Barrier-free Living Environment Certification Standards - A Focus on Boramae Park- (장애물 없는 생활환경 인증기준을 적용한 도시공원 개선계획 - 보라매공원을 대상으로 -)

  • Seo, Eun-Sil;Koo, Bon-Hak
    • Journal of the Korean Institute of Landscape Architecture
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    • v.43 no.5
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    • pp.100-110
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    • 2015
  • With the trend for the construction of barrier-free living environments in accordance with the ever-increasing external activity of the aging population, multi-culture families and socially disadvantaged, there is also a demand for the facilities improvement at parks for their convenience of use. This study analyzed the possibility of Barrier-Free Certification of an existing open space and defined its prototype renovation plan. This study is intended to help develop Borame Park into a barrier-free park by securing a barrier-free line of movement linking the major facilities sites within the park, improving park facilities lest there should be inconvenience in the use of the major facilities, and a guidance system convenient for use by physically and mentally impaired people. The significance of this study lies in the fact that it diagnosed the possibility of the BF Certification of existing urban parks and established a barrier-free park, which is easy for anyone to use. In the course of doing research on this, this study was able to discover the limitations of the BF Certification system, and its related tasks to be solved in the future as follows: The first thing this study shows is that there is a lack of understanding and awareness of the BF Certification system. Basically, there is a need for giving consideration to the development of a barrier-free living environment from the planning and design phase of a park, but in actuality, an examination of the certification is undertaken at the time of the completion of design and construction, which causes a large loss both time-wise and economically. Second, as evaluative items for the BF Certification of a park, applied are detailed evaluation items that fail to reflect the characteristics of an outdoor space whose natural constraints are large, such as geographical highs and lows and existing trees, plants, etc. Third, guidelines for establishment of a barrier-free park must be created in the light of the locational conditions of the park, followed by improvements of its BF Certification evaluation items.

Applications of Fuzzy Theory on The Location Decision of Logistics Facilities (퍼지이론을 이용한 물류단지 입지 및 규모결정에 관한 연구)

  • 이승재;정창무;이헌주
    • Journal of Korean Society of Transportation
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    • v.18 no.1
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    • pp.75-85
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    • 2000
  • In existing models in optimization, the crisp data improve has been used in the objective or constraints to derive the optimal solution, Besides, the subjective environments are eliminated because the complex and uncertain circumstances were regarded as Probable ambiguity, In other words those optimal solutions in the existing models could be the complete satisfactory solutions to the objective functions in the Process of application for industrial engineering methods to minimize risks of decision-making. As a result of those, decision-makers in location Problems couldn't face appropriately with the variation of demand as well as other variables and couldn't Provide the chance of wide selection because of the insufficient information. So under the circumstance. it has been to develop the model for the location and size decision problems of logistics facility in the use of the fuzzy theory in the intention of making the most reasonable decision in the Point of subjective view under ambiguous circumstances, in the foundation of the existing decision-making problems which must satisfy the constraints to optimize the objective function in strictly given conditions in this study. Introducing the Process used in this study after the establishment of a general mixed integer Programming(MIP) model based upon the result of existing studies to decide the location and size simultaneously, a fuzzy mixed integer Programming(FMIP) model has been developed in the use of fuzzy theory. And the general linear Programming software, LINDO 6.01 has been used to simulate, to evaluate the developed model with the examples and to judge of the appropriateness and adaptability of the model(FMIP) in the real world.

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ICM - Trophectoderm Cell Numbers of Mouse IVF/IVC Blastocysts (체외생산된 생쥐 배반포기배의 ICM과 Trophectoderm 세포수에 관한 연구)

  • Kim, E.Y.;Kim, S.E.;Uhm, S.J.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.1
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    • pp.25-32
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    • 1996
  • This work has been carried out to examine the number of Total, ICM and TE cells of F1 mouse blastcysts at day 4 after IVF by differential labelling of the nuclei with polynucleotide-specific fluorochromes and to obtain a fundamental information of preimplantation mouse embryo development. Blastocysts produced by superovulated B6CBA F1(C57BL/${\times}$CBA) eggs were inseminated with $1{\times}10^6$spermatozoa/ml and cultured in M16 medium at $37^{\circ}C$, 5% $CO_2$ incubator for 95hrs. Blastocysts were classified as early, middle, expanded and hatching stage according to the developmental morphology; blastocoel expansion and zona thickness. The results obtained in these experiments were summarized as follows; 1) The development rate of blastocysts at 95hrs after IVF was 86.7% and classified blastocysts to early, middle, expanded and hatching were 16.3%, 18.9%, 10.5% and 40.9%, respectively. 2) The numbers of total blastomere using bisbenzimide in the classified blastocysts to early, middle, expanded and hatching were 35.6${\pm}$1O.4, 49.4${\pm}$8.6, 60.8${\pm}$1O.7 and 62.7${\pm}$13.9, respectively. 3) In ICM and TE cell number by using differential labelling with polynucleotide-specific fluorochrome in the classified blastocysts to early, middle, expanded and hatching; ICM numbers were 9.6${\pm}$3.0, 13.6${\pm}$3.9, 16.0${\pm}$3.3 and 19.5${\pm}$4.6, respectively and TE cell numbers were 30.6${\pm}$5.1, 39.9${\pm}$5.8, 42.2${\pm}$8.1 and 43.7${\pm}$11.1, respectively. These results showed the same increase pattern according to development advance level. Also, when compared with the results of total count were obtained between bisbenzimide only and differential labelling, both of them showed the same increase pattern according to development level and at the same time their cell numbers were almost the same. So, rapid and simple cell count method using differential labelling can be used for the examination of later preimplantation development or as an indicator of embryo quality according to the variables of culture conditions.

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Studies on the Use of Radioisotope Tracer Techniques to Investigate and Improve the Root Activities in Rice Plant(I) - Effect of Water Control in Soil of the Paddy Field Lacking in the Special Mineral Nutritions - (방사성(放射性) 동위체도입(同位體導入)과 그 추적기술(追跡技術)에 의(依)한 수도근계(水稻根系) 활성상(活性相)의 해명(解明)과 개선(改善)에 관(關)한 연구(硏究) - 특수성분(特殊成分) 결핍(缺乏) 답토양(沓土壤)에서의 용수조절(用水調節) 효과(效果)에 대(對)하여-(제1보)(第1報) -)

  • Ahn, Hak-Soo;Chung, Hee-Don;Kim, Kyu-Won;Shim, Sang-Chil
    • Applied Biological Chemistry
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    • v.15 no.1
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    • pp.77-84
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    • 1972
  • A field experiment was conducted to determine the factors responsible in limiting nutrient up take and root activity in low productive paddy fold. Radiosotope of phosphorus-32 was used as a tracer. Results of the study were as follows: 1. On yield components responsible for increase yield indicated that number of ears per panicle and ripening ratio were closely related to increase yield. 2. Root volume or root feeding area has significant influence in increasing rice yield. 3. Root volume indicative of root activity and nutrient uptake can be effected by reasonable water control. 4. The combined application of calcium, silica, and magnesium(as a fused magnesium phosphate. the Kyun-gi Chemical Co. products.) with water control, although under conditions of large amount application of nitrogen, was found to be increased the maturing rate. 5. In the plots of water control, the number of roots per one volume were less than that of the continuous flooding plots, but the weight per root was heavier than the flooding plot ones. 6. Improvement of the present native culture method could effectively increase paddy rice yield.

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The Optimal Production and Characteristics of an Alginate-degrading Enzyme from Vibrio sp. PKA 1003 (Vibrio sp. PKA 1003의 알긴산 분해 조효소 생산 최적 조건과 조효소의 특성)

  • Kim, Hyun-Jee;Kim, Koth-Bong-Woo-Ri;Kim, Dong-Hyun;SunWoo, Chan;Jung, Seul-A;Jeong, Da-Hyun;Jung, Hee-Ye;Lim, Sung-Mee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.434-440
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    • 2013
  • This study was conducted to screen the characteristics and alginate-degrading activity of marine bacterium isolated from brown seaweed (Sargassum thunbergii). The results of 16S ribosomal RNA sequence analysis the strain the genus Vibrio and the strain was subsequently named Vibrio sp. PKA 1003. The optimum culture conditions for the growth of Vibrio sp. PKA 1003 were at pH 7, 3% NaCl, $25^{\circ}C$, and 6% alginic acid, with a 48-hour incubation time. A crude enzyme preparation from Vibrio sp. PKA 1003, showed its highest levels of alginate-degradation activity when cultured at pH 9, $30^{\circ}C$, and 6% alginic acid, with a 63-hour incubation time. Thin layer chromatography analyses confirmed that the crude enzyme released monomers or oligomers from sodium alginate, and results from trypsin treatment showed that the alginate degrading activity depends on this enzyme produced by Vibrio sp. PKA 1003. These results suggest that Vibrio sp. PKA 1003 and its alginate-degrading crude enzyme is useful for the production of alginate oligosaccharides.

Mouse Embryonic Stem Cell Uptakes of Buforin 2 and pEP-1 Conjugated with EGFP (생쥐 배아 줄기세포의 Buforin 2 및 pEP-1에 결합된 EGFP의 세포 내 수송)

  • Jung, Su-Hyun;Park, Seong-Soon;Lim, Hyun-Jung;Cheon, Yong-Pil
    • Development and Reproduction
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    • v.11 no.2
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    • pp.111-119
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    • 2007
  • Differentiation of cells can be induced through modulation of endogenous regulators using exogenous factors. Useful transfection systems to transport a specific exogenous regulator into cell have been tried but still there are many obstacles to overcome. In this study, we examined the transfection efficiency of cell permeable peptides (CPPs) in mouse embryonic stem cell under the various conditions. To identify the CPP-mediated translocation of a protein, we employed recombinant CPP-enhanced green fluorescent protein (EGFP). Viability of R1 cells was different between experimental groups depending on the kind of CPP and the concentration of CPP-EGFP. Translocation of CPP-EGFPs into the R1 cells was not detected until 30 min after CPP-EGFPs treatment in all groups. After 1 hr, translocation of pEP-1-EGFP-N was detected, but it could not be detected in the other group. Transfection of pEP-1EGFP-N was independent on its concentration. The time course did not show saturation even after 24 hr in pEP-1-EGFP-N. These results showed that the permeability depended on the kind of CPP and the location of His-tag in the case of examined CPPs, and did not need biological energy. On summary, the efficiency of transfection of CPP-EGFP depends on the CPP sequences but the culture time is not a key factor in transfection for the mouse embryonic stem cell. For the future studies to improve the efficiency of translocation of protein into embryonic stem cells, it is needed to develop modified CPP or mediator. The studies would be very useful to induce the differentiation of embryonic stem cells.

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Analysis on Growth and Yield of Cherry Tomato Grown in a Two-Story Bed System Adapted to Strawberry Cultivation as Affected by the Planting Time during the Uncultivated Period (딸기 재배용 2단 베드 시스템에서 휴작기 이용 방울토마토 재배 시 정식 시기에 따른 생육과 생산성 분석)

  • Choi, Hyo Gil;Moon, Byoung Yong;Kang, Nam Jun;Ko, Dae Whan;Kwon, Joon Kook;Lee, Jae Han;Park, Kyoung Sub
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.228-235
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    • 2016
  • This study was conducted to determine the yield of cherry tomato (Solanum lycopersicum Mill.) grown at three planting times during the uncultivated period of strawberry. Cherry tomato was planted under condition filled with strawberry dedicated culture medium on a two-story bed with April 20, April 30, and May 10 at 2015. Fruit harvest was completed on July 31. The supply concentration of nutrient solution at the time of transplanting was started as EC $1.2dS{\cdot}m^{-1}$ and it was gradually increased to EC $2.5dS{\cdot}m^{-1}$ after blooming of the first flower cluster. Netherlands PBG solution was supplied for one minute six times per day. The heights of cherry tomato plants planted at earlier were significantly greater than those of tomato plants planted later. The heights of cherry tomato plants grown at the bottom of the bed were greater than those grown in the upper bed. The yield of cherry tomatoes planted on April 20 at the bottom of the bed was greatest with an average of 2,954 g of tomatoes per plant. There were no significant differences in the average weight and sugar content of fruit according to planting times and bed position. The yield of cherry tomato plants planted on April 20 was 18% and 34% higher than that of plants planted on April 30 and May 10. We confirmed to increase the yield of the cherry tomato when early plants planted on two-story bed. These results indicate that farmers can choose the best period of producing cherry tomato during the un-cultivated period of strawberry under two-story bed conditions.

Production and Characterization of Extracellular Polysaccharide Produced by Pseudomonas sp. GP32 (Pseudomonas sp. GP32에 의해 생산된 세포 외 다당류의 생산 및 특성)

  • Lee, Myoung Eun;Lee, Hyun Don;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.25 no.9
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    • pp.1027-1035
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    • 2015
  • A strain GP32 which produces a highly viscous extracellular polysaccharide was conducted with soil samples and identified as Pseudomonas species. The culture flask conditions for the production of extracellular polysaccharide by Pseudomonas sp. GP32 were investigated. The most suitable carbon and nitrogen source for extracellular polysaccharide production were galactose and (NH4)2SO4. The optimum carbon/nitrogen ratio for the production of extracellular polysaccharide was around 50. The optimum pH and temperature for extracellular polysaccharide production was 7.5 and 32℃, respectively. In batch fermentation using a jar fermentor, the highest extracellular polysaccharide content (15.7 g/l) was obtained after 70 hr of cultivation. The extracellular polysaccharide produced by Pseudomonas sp. GP32 (designated Biopol32) was purified by ethanol precipitation, cetylpyridinium chloride (CPC) precipitation, and gel permeation chromatography. Biopol32, which has an estimated molecular weight of over 3×107 datons, is a novel polysaccharide derived from sugar components consisting of galactose, glucose, gulcouronic acid and galactouronic acid in an approximate molar ratio of 1.85 : 3.24 : 1.00 : 1.42. The solution of Biopol32 showed non-Newtonian characteristics. The viscosity of Biopol32 exhibited appeared to be higher at all concentration compared to that of zooglan from Zoogloea ramigera. An analysis of the flocculating efficiency of Biopol32 in industry wastewater (food, textile, and paper wastewater) revealed chemical oxygen demand (COD) reduction rates 58.4-67.3% and suspended solid (SS) removal rates 82.6-91.3%. Based on these results, Biopol32 is a possible candidate for industrial applications such as wastewater treatment.