• Title/Summary/Keyword: removal of cells

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Comparison of the Methods of Zona Pellucida Removal and Inner Cell Mass Isolation for the Generation of Parthenogenetic Embryonic Stem Cells in HanWoo Cattle

  • Kim, Dae-Hwan;Park, Sang-Kyu;Kim, Se-Woong;Jung, Yeon-Gil;Roh, Sang-Ho
    • Journal of Embryo Transfer
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    • v.26 no.2
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    • pp.111-115
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    • 2011
  • In general, zona pellucida (ZP) of the blastocyst has to be removed first, then either isolated the inner cell mass (ICM) or ZP-removed whole blastocyst, which is then cultured on the feeder layer to induce ICM outgrowth for the generation of embryonic stem cells (ESC). However, it is unclear whether ICM isolation before seeding on feeder layer is beneficial or not because the interaction between ICM and trophoblasts may affect cellular growth and/or pluripotency during the culture on the feeder. In the present study, two ZP removal methods (mechanically by splitting with a 28-gauge needle versus chemically by the treatment of acid-Tyrode's solution) and two ICM isolation methods (ZP-free whole blastocyst seeding versus mechanical isolation of ICM) were evaluated for the efficient isolation and culture of putative parthenogenetic bovine ESC. The number of maintained outgrown colonies was counted in each experimental group. As the result, mechanical removal of ZP with a needle and followed by whole ZP-free blastocyst seeding on feeder cells tended to attach more on the feeder layer and resulted in more outgrown colonies with its simple and less time-costing benefits. Currently we are generating ESC lines in HanWoo cattle by using this method for initial outgrowth of the parthenogenetic bovine blastocysts.

The Effect of Dimethyl-Sulfoxide and Sucrose as a Cryoprotectant on the Adenosine Triphosphate and Ultrastructure of Bovine Oocytes Matured In Vitro

  • Tsuzuki, Yasuhiro;Hisanaga, Mio;Ashizawa, Koji;Fujihara, Noboru
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.10
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    • pp.1353-1359
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    • 2001
  • The present study was undertaken to assess the influence of dimethyl-sulfoxide plus sucrose solution as a cryoprotectant on the adenosine triphosphate (ATP) content, the ultrastructure and the embryonic development of bovine oocytes matured in vitro. We measured the amount of ATP in cumulus cells enclosed oocytes (CO) or denuded oocytes (DO) equilibrated with or removed from the cryoprotectant (1.5 M DMSO + 0.25 M sucrose + 20% fetal bovine serum in physiological saline). As a result, the ATP contents in both CO and DO, equilibrated with the cryoprotectant, were significantly lower (p<0.05) than that of the each control group. However, ATP content of DO was recovered to the level of the control group ailer removal of the cryoprotectant, but failed to restore for CO. In the observation of the ultrastructure by a transmission electron microscope, all of the mitochondria in the ooplasm of CO and DO equilibrated with the cryoprotectant were swollen with peripherally located cristae following decreased electron density. However, a large proportion of these swollen mitochondria were restored to the normal shape which can be observed usually in the control group after removal from the cryoprotectant. To the contrary, the morphology of many mitochondria of the cumulus cells in CO were not recovered to that of the control group after removal of the cryoprotectant. CO with removed cryoprotectant had significantly lower embryonic development up to the blastocysts stage (p<0.05) after in vitro fertilization compared with that in the control group. These results suggest that the addition and removal of a cryoprotectant has a negative effect for the ATP content of cumulus enclosed oocytes. One of the factor(s) causing the lower embryonic development after removal of cryoprotectant, may be associated with ATP metabolism.

Microbial Community Analysis of 5-Stage Biological Nutrient Removal Process with Step Feed System

  • Park, Jong-Bok;Lee, Han-Woong;Lee, Soo-Youn;Lee, Jung-Ok;Bang, Iel-Soo;Park, Eui-So;Park, Doo-Hyun;Park, Yong-Keun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.929-935
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    • 2002
  • The 5-stage biological nutrient removal (BNR) process with step feed system showed a very stable organic carbon and nutrient removal efficiency ($87\%\;COD\,;79\%\;nitrogen,\;and\;87\%$ phosphorus) for an operation period of 2 years. In each stage at the pilot plant, microbial communities, which are important in removing nitrogen and phosphorus, were investigated using fluorescence in-situ hybridization (FISH) and 165 rDNA characterization. All tanks of 5-stage sludge had a similar composition of bacterial communities. The totat cell numbers of each reactor were found to be around $2.36-2.83{\times}10^9$ cells/ml. About $56.5-62.0\%$ of total 4,6-diamidino-2-phenylindol (DAPI) cells were hybridized to the bacterial-specific probe EUB388. Members of ${\beta}$-proteobacteria were the most abundant proteobacterial group, accounting for up to $20.6-26.7\%$. The high G+C Gram-positive bacterial group and Cytophaga-Flexibacter cluster counts were also found to be relatively high. The beta subclass proteobacteria did not accumulate a large amount of polyphosphate. The proportion of phosphorus-accumulating organisms (PAOs) in the total population of the sludge was almost $50\%$ in anoxic-1 tank. The high G+C Gram-positive bacteria and Cytophaga-Flexibacter cluster indicate a key role of denitrifying phosphorus-accumulating organisms (dPAOs). Both groups might be correlated with some other subclass of proteobacteria for enhancing nitrogen and phosphorus removal in this process.

Biosorptive capacity of Cd(II) and Pb(II) by lyophilized cells of Pleurotus eryngii

  • Joo, Jin-Ho;Hussein, Khalid A.;Hassan, Sedky H.A.
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.4
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    • pp.615-624
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    • 2011
  • last few decades. In this study, the lyophilized cells of Pleurotus eryngii (mushroom) were used as an inexpensive biosorbent for Cd(II) and Pb(II) removal from aqueous solutions. The effect of various physicochemical factors on Cd(II) and Pb(II) biosorption such as pH (2.0-7.0), initial metal concentration ($0.0-300mg\;L^{-1}$), temperature, fungal biomass and contact time (0-120 min) were studied. Optimum pH for removal of Cd(II) and Pb(II) was 6.0, and the contact time was 45 min at room temperature. The nature of biosorbent and metal ion interaction was evaluated by Infrared (IR) spectroscopic technique. IR analysis of mushroom biomass revealed the presence of amino, carboxyl, hydroxyl and methyl groups, which are responsible for biosorption of Cd(II) and Pb(II). The maximum adsorption capacities of P. eryngii for Pb(II) and Cd(II) calculated using Langmuir adsorption isotherm were 82.0 and $16.13mg\;g^{-1}$, respectively. The adsorption isotherms for two biosorbed heavy metals were fitted well with Freundlich isotherm as well as Langmuir model with correlation coefficient ($r^2$>0.99). Thus, this study indicated that the P. eryngii is an efficient biosorbent for the removal of Cd(II) and Pb(II) from aqueous solutions.

Effect of Extracellular Polymeric Substances(EPS) on the Biosorption of Lead by Microorganisums (납의 생물흡착에 미치는 세포외고분자물질의 영향)

  • 서정호;김동석;송승구
    • KSBB Journal
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    • v.14 no.1
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    • pp.66-70
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    • 1999
  • Comparison of lead removal characteristics between two strains, Aureobasidium pullulans and Saccharomyces cerevisiae, and effects of extracellular polymeric substances(EPS) excreted by microorganisms on the removal of lead were investigated. The capacity of lead biosorption to A. pullulans which had EPS was increased as the storage time of the cells increased, due to the increased amounts of excreted EPS. When the EPS were removed from A. pullulans cells, the amounts of adsorbed lead were very small(10% of the cell with EPS). In the case of s. cerevisiae which had no EPS, the lead removal capacity was nearly constant with storage time except early stage, but the spending time to reach an equilibrium state decreased with increasing storage time because of lowering the function of cell membrane. Therefore, it seems that the phenomena of lead biosorption were remarkably affected by the presence of extracellular polymeric substances.

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Investigation of Wet Chemical Etching for Surface Texturing of Multi-crystalline Silicon Wafers (다결정 실리콘 웨이퍼의 표면 텍스쳐링을 위한 습식 화학 식각에 대한 연구)

  • Kim, Bum-Ho;Lee, Hyun-Woo;Lee, Eun-Joo;Lee, Soo-Hong
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2006.11a
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    • pp.19-20
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    • 2006
  • Two methods that can reduce reflectance in solar cells are surface texturing and anti-reflection coating. Wet chemical etching is a typical method that surface texturing of multi-crystalline silicon. Wet chemical etching methods are the acid texturization of saw damage on the surface of multi-crystalline silicon or double-step chemical etching after KOH saw damage removal too. These methods of surface texturing are realized by chemical etching in acid solutions HF-$HNO_3$-$H_2O$. In this solutions we can reduce reflectance spectra by simple process etching of multi-crystalline silicon surface. We have obtained reflectance of 27.19% m 400~1100nm from acidic chemical etching after KOH saw damage removal. This result is about 7% less than just saw damage removal substrate. The surface morphology observed by microscope and scanning electron microscopy (SEM).

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Activity of Chlorelaa vulgaris Associated by Escherichia coli W3110 on Removal of Total Organic Carbon in Continuous River Water Flow System

  • Kong, Surk-Key;Nakajima Toshiuki
    • ALGAE
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    • v.17 no.3
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    • pp.195-199
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    • 2002
  • We investigated the association of Chlorella vulgaris and E. coli W9110 in removal of total organic carbon with the lab-scaled continuous river water flow system (CRWFS). Artificial wastewater was applied at two levels of organic carbon concentration; 1,335 $mg{\cdot}l^{-1}$ in the treatment (T)-1 and 267 $mg{\cdot}l^{-1}$ in T-2. The highest densities of C. vulgaris were $8.3{\times10^6\;cells{\cdot}ml^{-1}$ in T-1 and $6.9{\times}10^6\;cells{\cdot}ml^{-1}$ in T-2. The maximum densities of E. coli W3110 were $2.0{\times}10^8$ clony forming unit (CFU)${\cdot}ml^{-1}$ in T-1 and $3.9{\times}10^8\;CFU{\cdot}ml^{-1}$ in T-2. The densities increased during the first 11 days in T-q and 4 days in T-2, and decreased rapidly till 35th day, then increased slightly afterwards. This trend was prominent in T-2. It was inplied that wider range of nutrients was required in the growth of heterotrophic bacteria in T-2 than in T-1. The algal biomass should be increased effectively for the successful removal of organic carbon.

백서악하선적출이 갑상선에 미치는 영향

  • Cheong, Dong-Kyun;Park, No-Hee;Kim, Byoung-Gill;Kim, Chang-Whe
    • The Journal of the Korean dental association
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    • v.12 no.12
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    • pp.925-927
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    • 1974
  • It has been said that thyroid gland controls the function of salivary gland and vice versa. In an attempt to investigate the role of submaxillary gland on the function of thyroid gland, the authors have observed the changes of thyroid gland histologically and histochemically at 70 days after removal of submaxillary gland in rats. The results are as follows: 1. There are no significant changes of thyroid cells after submaxillary gland removal. 2. The colloidal concentration of thyroid follicles is markedly decreased after submaxillary gland removal. 3. The colloids of thyroid follicles show intensive reaction to PAS staining in intact rats, but moderate or minimal reaction in submaxillary removal rats.

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Behavior of Indoor Organic Pollutants Dissolved into the Ringer's Solution (수액제에 유입되는 실내 유기오염물질의 거동)

  • 김만구;정영림
    • Journal of Korean Society for Atmospheric Environment
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    • v.12 no.3
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    • pp.315-322
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    • 1996
  • Recently, hospital acquired infections have an increase interest as a public problems, which are caused of indoor pollutants in hospital. Microorganisms, ethylene oxide, formaldehyde, and anesthetic gases are main hazardous pollutants in hospital. The possible pathways of the infection are a respiratory channel as well as a blood channel. The blood channel is concerned since these pollutants might be dissolved into the Ringer's solution. The objective of this research was to evaluate the removal efficiencies of adsorption trap for formaldehyde and microorganisms as indoor pollutants which permeated into the Ringer's solution. Dissolved formaldehyde in the solution was increased with the injection dose time. The amount of dissolved formaldehyde was 67.5 $\pm$ 9.5% in Ringer's solution when injection dose time was controlled about 7hrs. An adsorption trap was designed for preventing formaldehyde and microorganisms to be permeated into Ringer's solution. The adsorption trap was packed with 0.4g of active carbon (60/80 mesh) in a sterilized plastic tube (7.79 cm length, 0.46 cm i.d.) and both ends were packed with glass wool. Devised infusion set equipped with the adsorption trap showed 99.9% of removal efficiency for formaldehyde. Microorganism numbers detected on sterilized water for injection and 5% dextrose infusion used in the hospital were 2,695 $\times 10^3$ cells/l and 4,190 $\times 10^3$ cells/l, respectively. Removal efficiency by the adsorption trap was 92.3 $\pm$ 8.5% as for microorgnisms.

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Continuous Deodorization of Malodorous Sulfur Compounds Using Immobilized Riohacillus neapolitanuts R-10 (고정화 Thiobacillus neapolitanus R-10를 이용한 유황계 악취물질의 연속제거)

  • 원용돈;박상보
    • Journal of Environmental Science International
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    • v.4 no.3
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    • pp.295-301
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    • 1995
  • Continuous deodorization of malodorous sulfur compounds by Thiobaillus neapolitanusts R-10 immobilized onto a polypropylene pellet was studied using a column reactor at 30$^{\circ}$C. The maximum amounts of immobilized cells was 5.3 gall polypropylene with 5$\times$7.5mm in pellet size, and the amounts of immobilized cells in the higher part of the column was as twice as in the lower part. The optimum pH and temperature for removal of dimethyl sulfide were 6.0 and 30$^{\circ}$C, respectively. When 5-20 ${mu}ell$/l of hydrogen sulfide and methylmercaptan were employed 98% of removal efficiency were achieved. In contrast, lower concentrations of dimethyl sulfide and dimethyldisulfide should be supplied to meet satisfactory deodorization efficiency. The immobilized cell column was successfully operated for the deodorization of mixture of sulfur compounds over 15 days without significant loss of initial activity achieving high efficiency.

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