• Title/Summary/Keyword: defined medium

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Comparison of Two Different Serum-free Media for In Vitro Culture of Bovine Embryos

  • Kim, Se-Woong;Jung, Yeon-Gil;Park, Jong-Im;Roh, Sangho
    • Journal of Embryo Transfer
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    • v.29 no.3
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    • pp.229-234
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    • 2014
  • The aim of the present study was to compare two different serum-free media, modified synthetic oviduct fluid (mSOF) and modified potassium simplex optimization medium (mKSOM) containing 20% RD (RPMI1640 + DMEM, 1:1 v/v) (RD-mKSOM), for in vitro culture (IVC) of bovine embryos. After in vitro maturation and fertilization, the presumptive zygotes were cultured in two different serum-free conditions for 7 days and 9 days to evaluate blastocyst formation and hatching, respectively. Serum supplemented conventional CR2 medium was used as control. After 7 day of culture, there was no significant difference in cleavage and blastocyst formation rates among three groups (mSOF, 59.3 and 30.1%; RD-mKSOM, 65.0 and 41.5%; control, 51.6 and 38.0%, respectively). Hatching rate was significantly higher in control (69.0%) than other experimental groups (mSOF, 22.0%; RD-mKSOM, 39.5%) (P<0.0001 and P<0.001, respectively). Although both serum-free conditions showed lower hatching rates than serum-added control, in serum-free groups, RD-mKSOM showed significantly higher hatching rate than mSOF (P<0.001). In addition, one-step using RD-mKSOM may facilitate IVC procedure than two-step culture system. In conclusion, the results indicate that one-step RD-mKSOM is more suitable defined culture system for IVC of bovine embryos than two-step mSOF.

The Production of Tissue Type Plasminogen Activator from Normal Human Cell tine (정상 인체 세포로부터 조직 플라스미노겐 활성인자의 대량생산)

  • Lee, Hyeon-Yong;Kim, Geum-Soo
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.522-525
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    • 1988
  • A method to produce tissue type Plasminogen Activator (tPA) from normal human fibroblast is developed by cultivating cells in serum free media containing heparin as an inducer. Optimal dose of this inducer was 30$\mu$g/m$\ell$. The composition of serum free medium was also defined to fit to the industrial scale cultivation. 1.42 ug of tPA per 10$^5$ viable cells per ml was produced. 1.1 gram of tPA can be produced every day from this cell line under normal perfusion chemostat operations assuming that same productivity is maintained when the process is sealed up. This method could reduce pro-duction costs and simplify purification processes by using serum free medium. Tissue type PA produced from this cell line has high ability of dissolving clots, based upon fibrin lysis test showing 50mm$^2$ of clearing zones in agarose gel plate. These results were reproducible and in good agreement with results of ELISA assay. tPA from normal human cells will be safer than that from melanoma and recombinant cells in human clinical trials.

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A Comparative Study for Grinding Media Behavior and DEM Simulation at Actual Grinding Zone on a Traditional Ball Mill (매체형 전동밀의 실제 분쇄장에서 분쇄매체의 거동과 DEM 시뮬레이션의 비교연구)

  • Bor, Amgalan;Jargalsaikhan, Battsetseg;Lee, Jehyun;Choi, Heekyu
    • Korean Chemical Engineering Research
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    • v.57 no.6
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    • pp.804-811
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    • 2019
  • This study was performed the real motion and DEM simulation of ball motion using three different types of grinding media with different size and materials in media formation for optimization of process conditions in a traditional ball mill (media ball mill). In the simulation, the rotational speed of the mill, the material of the medium, the velocity of the medium, and the coefficient of friction between the media and the wall of pot were fixed into the actual experimental conditions. The motion of various kinds of grinding media was quantitatively measured by setting the grinding zone defined in the present study on the photographs taken and the snapshot images analyzed in the simulation. In addition, we observed the quantitatively measured value and the changed morphology of the sample and examined the correlation. Therefore, it is suggested to optimize the grinding media which has the greatest influence on the grinding zone under specific experimental conditions.

Increased cryo-survival rate in ejaculated human sperm from infertile men following pre-freeze in vitro myo-inositol supplementation

  • Saleh, Ramadan;Assaf, Hanan;Abd El Maged, Wafaa M.;Elsuity, Mohamed;Fawzy, Mohamed
    • Clinical and Experimental Reproductive Medicine
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    • v.45 no.4
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    • pp.177-182
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    • 2018
  • Objective: To investigate the effects of in vitro myo-inositol (Myo-Ins) supplementation of cryopreserved human semen on the cryo-survival rate (CSR). Methods: Semen samples were obtained from 41 infertile men. Following routine semen analysis, each sample was divided into two equal aliquots (0.5 mL each). One aliquot was treated with 1 mg of Myo-Ins dissolved in $10{\mu}L$ of sperm preparation medium. The second aliquot was treated with $10{\mu}L$ of the same medium (control). Both aliquots were incubated for 20 minutes prior to freezing to slow the freezing process. The frozen samples were examined for post-thaw percentages of total motility (TM), progressive motility (PM), and the CSR, defined as the percentage of post-thaw TM divided by the percentage of pre-freeze TM and multiplied in 100. The results were expressed as median and interquartile range (25th and 75th percentiles). Results: The pre-freeze TM (50% [30%-50%]) and PM (35% [20%-35%]) were significantly higher than the post-thaw TM and PM in the MyoIns group (15% [10%-35%] and 10% [5%-20%]; p<0.001 and p<0.001, respectively) and the control group (10% [6%-30%] and 5% [3%-15%]; p<0.001 and p<0.001, respectively). The CSR of the 41 semen aliquots supplemented with Myo-Ins (40% [25%-70%]) was significantly higher than that of the control samples (30% [13%-58%], p=0.041). The CSR of the 26 abnormal semen samples that were supplemented with Myo-Ins (38% [20%-50%]) was significantly higher than that of the control samples (23% [12%-30%], p=0.031). Conclusion: In vitro Myo-Ins supplementation of ejaculated human sperm from infertile men resulted in a significant increase in the CSR in samples with abnormal pre-freeze sperm parameters.

Fundamental Research for Video-Integrated Collision Prediction and Fall Detection System to Support Navigation Safety of Vessels

  • Kim, Bae-Sung;Woo, Yun-Tae;Yu, Yung-Ho;Hwang, Hun-Gyu
    • Journal of Ocean Engineering and Technology
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    • v.35 no.1
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    • pp.91-97
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    • 2021
  • Marine accidents caused by ships have brought about economic and social losses as well as human casualties. Most of these accidents are caused by small and medium-sized ships and are due to their poor conditions and insufficient equipment compared with larger vessels. Measures are quickly needed to improve the conditions. This paper discusses a video-integrated collision prediction and fall detection system to support the safe navigation of small- and medium-sized ships. The system predicts the collision of ships and detects falls by crew members using the CCTV, displays the analyzed integrated information using automatic identification system (AIS) messages, and provides alerts for the risks identified. The design consists of an object recognition algorithm, interface module, integrated display module, collision prediction and fall detection module, and an alarm management module. For the basic research, we implemented a deep learning algorithm to recognize the ship and crew from images, and an interface module to manage messages from AIS. To verify the implemented algorithm, we conducted tests using 120 images. Object recognition performance is calculated as mAP by comparing the pre-defined object with the object recognized through the algorithms. As results, the object recognition performance of the ship and the crew were approximately 50.44 mAP and 46.76 mAP each. The interface module showed that messages from the installed AIS were accurately converted according to the international standard. Therefore, we implemented an object recognition algorithm and interface module in the designed collision prediction and fall detection system and validated their usability with testing.

Prediction of successful caudal epidural injection using color Doppler ultrasonography in the paramedian sagittal oblique view of the lumbosacral spine

  • Yoo, Seon Woo;Ki, Min-Jong;Doo, A Ram;Woo, Cheol Jong;Kim, Ye Sull;Son, Ji-Seon
    • The Korean Journal of Pain
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    • v.34 no.3
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    • pp.339-345
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    • 2021
  • Background: Ultrasound-guided caudal epidural injection (CEI) is limited in that it cannot confirm drug distribution at the target site without fluoroscopy. We hypothesized that visualization of solution flow through the inter-laminar space of the lumbosacral spine using color Doppler ultrasound alone would allow for confirmation of drug distribution. Therefore, we aimed to prospectively evaluate the usefulness of this method by comparing the color Doppler image in the paramedian sagittal oblique view of the lumbosacral spine (LS-PSOV) with the distribution of the contrast medium observed during fluoroscopy. Methods: Sixty-five patients received a 10-mL CEI of solution containing contrast medium under ultrasound guidance. During injection, flow was observed in the LSPSOV using color Doppler ultrasonography, following which it was confirmed using fluoroscopy. The presence of contrast image at L5-S1 on fluoroscopy was defined as "successful CEI." We then calculated prediction accuracy for successful CEI using color Doppler ultrasonography in the LS-PSOV. We also investigated the correlation between the distribution levels measured via color Doppler and fluoroscopy. Results: Prediction accuracy with color Doppler ultrasonography was 96.9%. The sensitivity, specificity, positive predictive value, and negative predictive value were 96.7%, 100%, 100%, and 60.0%, respectively. In 52 of 65 patients (80%), the highest level at which contrast image was observed was the same for both color Doppler ultrasonography and fluoroscopy. Conclusions: Our findings demonstrate that color Doppler ultrasonography in the LS-PSOV is a new method for determining whether a drug solution reaches the lumbosacral region (i.e., the main target level) without the need for fluoroscopy.

Daily Dose of Zizyphi Fructus in Treatise on Cold Damage (상한론(傷寒論)에서 대조(大棗)의 1 일 복용량)

  • Kim, In-Rak
    • The Korea Journal of Herbology
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    • v.28 no.1
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    • pp.51-58
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    • 2013
  • Objectives : As Daily Dose of Zizyphi Fructus was designated by the numbers in Treatise on Cold Damage Disease, estimation of Daily Dose was all different among researchers. The purpose of this study was to estimate exact Daily Dose of Zizyphi Fructus. Methods : We fixed the errors in various copy of Treatise on Cold Damage Disease and considered the meaning of the Bee Zizyphi Fructus(肥大棗) and general rules of Daily Dose in Treatise on Cold Damage Disease. So we reduced Daily Dose of Zizyphi Fructus, and compared this with the standard of Zizyphy Fructus in Pharmacopoeia of several Nation and Korean Forest Service. Results : Daily Dose of Zizyphi Fructus was generally 12 pieces; less was for prescriptions which scaled down the amount of ingredients prescribed in the originals; 15, 25, and 30 pieces were used when more was required. The medicinal part was the pulp of fructus, and the dosage of 12 pieces was respectively equivalent to 3 Ryang(兩), and 19.5 g. As defined in the Korean Pharmacopoeia Ninth Edition and standards of forest products by Korea Forest Service, Zizyphi Fructus was medium-sized, and weighs about 1.625 g if properly dehydrated. Conclusions : In short, the proper Daily Dose of Zizyphi Fructus in Treatise on Cold damage Disease was 12 pieces of Zizyphi Fructus and 19.5 g of its pulp, weighing three Ryang(兩). The pulp was estimated to be 1.625 g; it was medium-sized according to the present standard.

Study of Necessity of Advanced Integrated Digital Engineering and Management Tools (선진통합형 디지털 엔지니어링 및 경영 도구의 필요성 연구)

  • Luke (Yang Ouk) Kim;Kyung Ho Lee
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.36 no.1
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    • pp.27-38
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    • 2023
  • How the port, shipping, shipbuilding, and vender industries in Korea can have a seamless value chain through their digitally smooth cooperation by applying the latest information and communications technology through the "4th industrial revolution" was examined. Also considered was the proposition that the value chain should be a smart, seamless value chain among industries with successful hyperconnection. Their cooperative relationships were defined, and the crucial elements for the sustainable development of these industries were considered. As a result, the direction for achieving environmental, social, and governance management by realizing decarbonization through today's digitalization could be studied. In particular, the importance of digitization as a way to respond to the future market from the perspective of small and medium-sized enterprises and the role of digitization realized by small and medium-sized equipment companies in the overall industry were examined. The results simultaneously show the state of linkage between industries and the reason why the value chain must maintain a smooth relationship. In addition, using the lessons learned from recent failure cases from the Korean shipbuilding industry as a cornerstone, the direction for creating a strategic pathway for intelligent connection was investigated.

Studies on In Vitro Capacitation by Lysolecithin and In Vitro Fertilizing Ability of Ejaculated Rabbit Sperm

  • Kim, C.K.;Im, K.S.;Zheng, X.;Foote, R.H.
    • Korean Journal of Animal Reproduction
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    • v.10 no.1
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    • pp.109-120
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    • 1986
  • This study was conducted to define the effect of addition of lysolecithin (LC) and 20% v/v rabbit serum to sperm preincubation medium on the induction of acrosome reaction (AR) an fertilizing ability in vitro of LG-added sperm. Ejaculated rabbit sperm from New Zealand White buck was washed once by centrifugation, then preincubated for 2 or 4 hrs in a chemically defined medium (DM), DM plus 20% rabbit serum or BSA-free DM plus 20% rabbit serum at 37$^{\circ}C$ water bath or CO2 incubator. At the end of preincubation LC was added to the preincubated sperm, which was stained at 0.5 to 4 hr later and examined for AR and sperm motility. For in vitro fertilization, gametes were coincubated in DM up to 24 hrs and thereafter fertilized embryos were incubated in BSM -II up to 48 hrs. Addition of LC to 4-hr preincubated sperm was more effective for the AR and sperm motility than that to 2-hr preincubated sperm and optimal concentration of LC for AR was about 80${\mu}$g/ml. A significant increase in AR occured from 20 to 30 min. after addition of 80 to 100${\mu}$g/ml in 4-hr preincubated sperm. BSA-free DM plus 20% rabbit serum showed a higher AR and sperm motility than those of DM plus 20% rabbit serum in LC-added sperm after 4-hr preincubation. The incidence of AR after 4-hr preincubation and at 30 min after 60${\mu}$g/ml LC addition varied greatly among individual bucks. Sixty ${\mu}$g/ml LC-added sperm showed a slight high cleavage rate over control levels, but 100${\mu}$g/ml LC-added sperm showed lower cleavage rate rather than 60${\mu}$g/ml LC. It is concluded that optimal concentration of LC for high AR induction and sperm motility in 4-hr preincubated sperm was about 80${\mu}$g/ml, but 60${\mu}$g/ml level was more useful for in vitro fertilization.

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Effects of Development and Viability of Pig Oocytes Matured in Defined Medium Containing PVA, PVP and pFF (PVA, PVP 및 pFF를 첨가한 체외성숙 한정배지가 미성숙 돼지 난포란의 성숙과 배발달에 미치는 영향)

  • Kim I. D.;Kim S. N.;Han S. K.;Seok H. B.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.219-227
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    • 2004
  • This study was conducted to develop a serum-free, defined medium of IVM of pig oocytes. The TCM-199 with supplemented with polyvinylalcohol(PVA), polyvinylpyrrollidone(PVP) and porcine follicular fluid(pFF) were used as basal medium. The effects of the these additives on the rates of maturity and development under in-vitro fertilization and in vitro culture were examined and subsequently considered on the possibilities be sustituted for the bovine serum albumin(BSA). Maturation rate of pig oocytes in IVM media containing PVA(82.4%), pFF(89.4%) and BSA(90.0%) were significantly higher(P<0.05) than that of PVP(78.6%). Cleavage rate after IVF of PVP(64%) was significantly lower(P<0.05) than these of PVA(73%), pFF(77%) and BSA(73%) supplements. in vitro development rates to morulae and blastocyst on PVP(54%) were also significantly lower(P<0.05) than these of the supplements of PVA(63%), pFF(69%) and BSA(65%). In comparison of maturation and fertilization rates of pig oocytes in each supplements, the maturity rates of PVA(82.4%), pFF(89.4%) and BSA(90.0%) were significantly lower(P<0.05) than that of PVP(72.4%) and while, the fertilization rates of pFF(87.1%) and BSA(89.1%) were significantly higher(P<0.05) than these of PVA(78.0%) and PVP(70.6%). It may be concluded that PVA and pFF can be substituted far BSA in medium for culturing pig oocytes; however, it may be considered that PVP were limited to for BSA in the in vitro culture of the embryos.