• Title/Summary/Keyword: suspension volume

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Development of In Vitro Porcine Oocytes Following Intracytoplasmic Injection of Sperm-Mediated GFP Gene

  • Kim, J.H.;Seong, H.H.;Park, J.K.;Im, S.K.;Kim, S.W.;Lee, Y.K.;Lee, P.Y.;Choi, Y.J.;Kim, Y.K.;Kim, J.H.;Chang, W.K.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.69-69
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    • 2002
  • Transgenic animals production tools have been valuable for research and purpose. The current methods of gene transfer, microinjection and nuclear transfer, which are widely used in transgenic animal production, but all most methods has only had limited success in production of larger species. Here, we report the possibility of a sperm-mediated gene transfer method in porcine embryos. Oocytes were collected from ovaries harvested at a local slaughterhouse were matured in 500${mu}ell$ drops of TCM-199 under mineral oil at 38.5$^{\circ}C$ in a humidified atmosphere of 5%CO2 in air. After 42-43h of in vitro maturation oocytes were denuded. for sperm injection into the cytoplasm of the porcine oocytes, sperm suspension in NIM medium are subjected extraction with TritonX-100 before mixing with a green fluorescent gene (GFP). Sperm with Tritonx-100 were prepared by adding TritonX-100 to a final volume of 0.05% in the sperm suspension and mixing by trituration for 60s before two wishes in NIM medium at 2$^{\circ}C$. A(ter wishing, sperm were mixed with TritonX-100 at $25^{\circ}C$ followed by washes at 2$^{\circ}C$. Sperm were resuspended in ice cold NIM to a final volume of 400${mu}ell$ and 2-20ng/${mu}ell$ DNA were triturated on ice for 60s. All microinjection was performed in HEPES-buffered CZB medium at room temperature within 2h. After culture in NCSU-23 for 72h, percent of porcine embryos transfected GFP gene are 20.7%(6/29) in 20ng/${mu}ell$ sperm-DNA mixed group and other groups were 3.7 %(2/54)and 4.7%(3/67). These data suggests that sperm-mediated gene transfer method should be used to the production tool of transgenic pig efficiently.

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Effects of Mixing Performance and Conditioned Medium on hCTLA4Ig Production in Transgenic Rice Cell Suspension Cultures (형질전환 벼 현탁세포 배양에서 혼합효율과 조정배지가 hCTLA4Ig 생산에 미치는 영향)

  • Choi, Hong-Yeol;Park, Jun-Yong;Nam, Hyung-Jin;Gong, Mi-Kyung;Yoo, Ye-Ri;Kim, Dong-Il
    • KSBB Journal
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    • v.30 no.6
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    • pp.307-312
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    • 2015
  • Transgenic rice cells using RAmy3D promoter can provide high productivity, and the production of recombinant protein is induced by sugar starvation. In this system, productivity was reduced during the scale-up processes. To ensure the influences of shear stress and oxygen transfer rate, working volume and mixing performances were investigated under various agitation speeds and working volumes. In addition, inoculation methods including suspended cells and filtered cells were compared. Working volumes and shaking speeds were 300, 450 mL and 80, 120 rpm, respectively. Hydrodynamic environment of each condition was measured numerically like mixing time and $k_La$. Good mixing performance and high shear stress were measured at high agitation speed and low volume. The highest level of hCTLA4Ig was 30.7 mg/L at 120 rpm, 300 mL. When conditioned medium was used for inoculation, increased cell growth was noticed during the day 0~4 and decreased slower than filtered cells. Compared with filtered cells, the maximum hCTLA4Ig level reached 37.8 mg/L at 120 rpm, 300 mL and lower protease activity level was observed. In conclusion mixing performance is critical factor for productivity and conditioned medium can have a positive effect on damaged cells caused by hydrodynamic shear stress.

Evaluation of Antitumor and Antioxidant Activity of Sargassum tenerrimum against Ehrlich Ascites Carcinoma in Mice

  • Patra, Satyajit;Muthuraman, Meenakshi Sundaram;Prabhu, A.T.J. Ram;Priyadharshini, R. Ramya;Parthiban, Sujitha
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.915-921
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    • 2015
  • Context: In the last half century, discovering, developing and introducing of clinical agents from marine sources have seen great successes, with examples including the anti-cancer compound trabectedin. However, with increasing need for new anticancer drugs, further exploration for novel compounds from marine organism sources is strongly justified. Objective: The major aim of this study was to evaluate the antitumor and antioxidant potential of Sargassum tenerrimum J.Agardh (Sargassaceae) on Ehrlich ascites carcinoma (EAC) in Swiss albino mice. Materials and Methods: An ethanol extract of S. tenerrimum (EEST) from whole algae was used to evaluate cytotoxicity followed by in vivo assessment of toxicity, using biochemical parameters including hepatic and non-hepatic enzymes. Antioxidant properties were examined in animals bearing EAC treated with daily oral administration of 100-300 mg/kg extract suspension. Results: Antitumor effects of EEST in EAC bearing mice was observed with LD50 1815 mg/kg. Parameters like body weight, tumor volume, packed cell volume, tumor cell count, mean survival time and increase in life span in animals in the EAC bearing animals treated with EEST 300 mg/kg was comparable with control group. Significant differences were also seen with changes in total protein content, hepatic enzymes contents, MDA level, and free radical scavenging enzymes in untreated vs. EEST treated group animals. Conclusions: Evaluation of antioxidant enzymes and hepatic enzymes in the EAC animal model treated with EEST exhibited similar effects as the positive control drug 5-flurouracil. S. tenerrimum extracts contain effective antioxidants with significant antitumor activity.

Delivery System of Daunorubicin by Red Blood Cells (적혈구를 이용한 Daunorubicin의 배송시스템)

  • Ham, Seong-Ho;Song, Kyung;Ko, Gun-Il;Kim, Jae-Baek;Sohn, Dong-Hwan
    • Journal of Pharmaceutical Investigation
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    • v.24 no.3
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    • pp.131-137
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    • 1994
  • Drug delivery system by the use of red blood cells was established to sustain the release of drugs in the circulatory system by the intravenous injection. The entrapment method by the preswelling technique was re-examined and evaluated for searching the new entrapping conditions without hemolysis. The addition of 4 volume of $0.6{\times}\;hank's$ balanced salt solution (HBSS) into 1 volume of 50% red blood cells suspension did not induce the hemolysis and change the hematocrit level in this experimental condition (within 15 min). Most of daunorubicin could be entrapped into red blood cells within 15 min. While the intracellular adenosine triphosphate (ATP) level followed by the entrapment was reduced to 86% of normal ATP level, the membrane fluidity and the shape factor of red blood cells were not altered. The release rate of daunorubicin from red blood cells was affected by the hemolysis under this condition. To maintain the intracellular ATP in red blood cells, the new reaction buffer was made With the addition of ATP and sodium pyruvate during the entrapment procedure because the hemolysis during the release test would reflect the loss of intracellular ATP that might result in the decrease of the viability in vivo. The addition of ATP raised the intracellular ATP level, which protect the hemolysis during the release test.

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Preparation and Evaluation of Ketoprofen-incorporated Solid Lipid Nanoparticles (SLN) (케토프로펜을 함유하는 고형 지질 나노파티클의 제조 및 평가)

  • Baek, Myoung-Ki;Lee, Sang-Young;Jee, Ung-Kil
    • Journal of Pharmaceutical Investigation
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    • v.26 no.4
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    • pp.245-256
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    • 1996
  • Solid lipid nanoparticles (SLN) have been developed as a new drug delivery system. Although many particulate drug carriers, such as microsphere, liposome, niosome, emulsion, etc. have been introduced, they have some disadvantage; low efficiency of incorporation and stability, lack of reproducibility, and so on. Meanwhile, SLN as a new drug delivery system is known to entrap rugs with a high efficiency and a good reproducibility. Moreover, small size SLN can circulate in blood for a prolonged time. Although many preparation methods were introduced, microfluidization method is recommended to be the most useful. This study was attempted to prepare and evaluate ketoprofen-incorporated SLNs (keto-SLN), which were prepared by two methods, ultrasonication and microfluidization. Keto-SLN was evaluated by measurement of particle size and zeta potential, efficacy of entrapment, sedimentation volume, in virto release pattern. The mean particle size was about $0.1\;{\mu}m$, and the size was dependent on the type and the amount of emulsifier. Zeta potential was negative, $-9{\sim}-13mV$ and entrapment efficacy was very high and stability was good for at least 60 days in the respect of particle size and sedimentation volume ratio. Analgesic effect was also determined as well as pharmacokinetic parameters. The former was comparable to that of that of ketoprofen loaded suspension (keto-sus) and the latter revealed that consistent with the delayed release of keto-SLN. $T_{max}$ was longer than keto-sus. Therefore, keto-SLN was favourable dosage forms in the field of drug delivery system such as anti-cancer, analgesics and anti-inflammatory agents.

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Cold Gelatinization of Naked Barley Starches (쌀보리 전분의 화학적 호화)

  • Park, Yang-Kyun;Lho, Il-Hwan;Kim, Kwan;Kim, Sung-Kon
    • Korean Journal of Food Science and Technology
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    • v.18 no.3
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    • pp.192-196
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    • 1986
  • Cold gelatinization of two naked burley (Songhak and Youngsan) starches in NaOH (0.14-0.21N) and KSC N (1.5-4.5M) was compared. Light transmittance of starch suspension in aqueous NaOH solution was higher in Youngsan starch than in Songhak starch. Viscosity of Youngsan starch was also higher at various starch concentrations (5-12.5%), however the critical concentration of NaOH for the gelatinization of Youngsan starch was lower by 0.1meq NaOH/g starch compared to Songhak starch. Two starches held a linear relationship between viscosity development rate and alkali concentration at a fixed starch concentration. The viscosity development rate of the starch was linearly decreased as starch concentration increased at a fixed alkali concentration. Gel volume of the starches was attained maximum at 2.5M KSCN, however Youngsan starch showed a higher volume and a less stability in KSCN than Songhak starch.

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Dilutant flow characteristics model of coarse particle suspensions with uniform size distribution

  • Ookawara, Shinichi;Ogawa, Kohei
    • Korea-Australia Rheology Journal
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    • v.15 no.1
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    • pp.35-41
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    • 2003
  • It is expected that particle size distribution of any portion obtained through screening, is of more uniform than that of the original mixture, typically following such as log-normal, Rosin-Rammler distributions and so on. In this study, therefore, a new relation between parameters of the uniform distribution and flow characteristics of the coarse particle suspensions is derived based on the continuous polydisperse model (Ookawara and Ogawa, 2002b), which is derived from the discrete polydisperse model (Ookawara and Ogawa,2002a). The derived model equation predicts a linear increase of viscosity with shear rate, viz., dilutant flow characteristics. Further, the increase of viscosity is expected to be proportional to the square of volume fraction of particles, and to show the linear dependency on density and average diameter of particles. It is also shown that the uniform distribution model includes additional term that expresses the effect of distribution width. For verification of the model, the experimental results of Clarke (1967) are cited as well as in our previous work for the monodisperse model (Ookawara and Ogawa,2000) since most parameters were varied independently in his work. It is suggested that the newly introduced term expands the applicable range compared with the monodisperse model.

Antagonistic activity of Streptomyces halstedii and S. violaceusniger In pepper anthracnose fungus Colletotrichum gloeosporioides

  • Park, Dae-Yong;Lim, Tae-Heon;Byeongjin Cha
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.96.2-97
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    • 2003
  • More than 1200 microorganisms were isolated from soil samples collected from various sources and localities. Among the isolates, 2 actinomyces (TH-04 and BA313) and 1 Bacillus sp. (CJ3) were selected as antagonists to pepper anthracnose fungus Colletotrichum gloeosporioides. These 3 isolates inhibitied mycelial growth of C gloeosporioides and the inhibition rates were over 70% on PDA. When the isolates were co-cultured with conidia of C. gloeosporioides in potato dextrose broth, conidial germination was severely inhibited and the inhibition rates of TH-04, BA313, and CJ3 at 24 hours were 75%, 72%, and 68%, respectively. The inhibition rates at n hours incubation were not much different from the rates at 24 hours. To check the activity on the plant, each isolate was mixed with equal volume of conidial suspension of C. gloeosporioides and wound-inoculated on green pepper fruit. After 6 days, the anthracnose lesions on the fruits inoculated with the mixture were much smaller than the lesions caused by the C. gloeosporioides itself. The lesion areas of TH-04 or BA313 treated pepper were less than 30% of the check. TH-04 and BA313 also showed antagonistic activity to Phytophthora spp. and Botrytis cinerea. By scanning electron microscopy and fatty acid analyses (MIDI), TH-04 and BA313 were identified to Streptomyces halstedii and S. violaceusniger, repectively.

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Effects of Ginseng Extract on Biological Cell Membrane and Artificial Lipid Monolayer (인삼추출물이 생체 세포막 및 artificial lipid monolayer에 미치는 영향)

  • Paik, Kwang-Sei;Lee, Chul-Young;Lee, Kyung-Nam;Song, Sun-Ok;Kang, Doo-Hee
    • The Korean Journal of Physiology
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    • v.10 no.1
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    • pp.7-14
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    • 1976
  • The Present study was conducted to investigate the effects of Ginseng extract on the tension-area curve for stearic acid monolayer. At the same time, the effects of Ginseng extract on osmotic and mechanical fragility of human red cells and histamine release from rabbit leukocytes were studied, The results are summarized as follows. 1. The Ginseng alcohol extract was found to expand liquid expanded phase of stearic acid monolayer, thus it is speculated that this agent may be acting as a surface active substance. 2. Osmotic hemolysis was inhibited by the Ginseng alcohol extract and the same effect was also observed in the presence of Ginseng saponin. However, the Ginseng alcohol extract was found to decrease hematocrit ratio of the RBC suspension, therefore, the inhibition of the osmotic hemolysis by this agent may be secondary effect to the reduced cell volume. 3. The mechanical hemolysis was also inhibited by the Ginseng alcohol extract but the inhibition was independent of changes in hematocrit ratio. 4. Histamine release from rabbit leukocytes was significantly increased in vitro in the presence of the Ginseng alcohol extract.(p<0.05)

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Microstructure and shear modulus in concentrated dispersions of bidisperse charged spherical colloids

  • Chun, Myung-Suk;Lee, Sangwoo;Lee, Tae-Seok;Cho, Jae-Seol
    • Korea-Australia Rheology Journal
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    • v.16 no.1
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    • pp.17-26
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    • 2004
  • We examine rigorous computations on microstructural as well as rheological properties of concentrated dispersions of bidisperse colloids. The NVT Monte Carlo simulation is applied to obtain the radial distribution function for the concentrated system. The long-range electrostatic interactions between dissimilar spherical colloids are determined using the singularity method, which provides explicit solutions to the linearized electrostatic field. The increasing trend of osmotic pressure with increasing total particle concentration is reduced as the concentration ratio between large and small particles is increased. From the estimation of total structure factor, we observe the strong correlations developed between dissimilar spheres. As the particle concentration increases at a given ionic strength, the magnitude of the first peak in structure factors increases and also moves to higher wave number values. The increase of electrostatic interaction between same charged particles caused by the Debye screening effect provides an increase in both the osmotic pressure and the shear modulus. The higher volume fraction ratio providing larger interparticle spacing yields decreasing high frequency limit of the shear modulus, due to decreasing the particle interaction energy.