• Title/Summary/Keyword: Media effect

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A Novel Chenodeoxycholic Derivative HS-1200 Enhances Radiation-induced Apoptosis in Human MCF-7 Breast Cancer Cells (담즙산 합성유도체(HS-1200)가 인체 유방암 세포주(MCF-7)에서 유도하는 방사선 감작 효과)

  • Lee Hyung Sik;Choi Young Min;Kwon Hyuk Chan;Song Yeon Suk
    • Radiation Oncology Journal
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    • v.22 no.2
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    • pp.145-154
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    • 2004
  • Purpose : To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. Materials and Methods : Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37$^{\circ}C$ with 5$\%$ CO$_{2}$ in air atmosphere. After removal of HS-1200, cells were irradiated with 2$\~$8 Gy X-ray, and then cultured Ii drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16$\mu$M of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40$\mu$M of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken. Results : Treatment of MCF-7 cells with 40$\mu$M of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential ($\Delta$$\psi$$_{m}$) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. Conclusion : We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio In HS-1200 co-treatment group underlies the increased radio sensitivity of MCF-7 cells. Further futures studies are remained elusive.

Effect of Cholesterol Supplementation in Freezing Medium on the Survival and Integrity of Human Sperm after Cryopreservation (콜레스테롤이 동결-해동 후 인간정자의 생존과 기능보존에 미치는 영향)

  • Lim, Jung-Jin;Sung, Su-Ye;Kim, Kye-Seong;Song, Seung-Hon;Lee, Woo-Sik;Yoon, Tae-Ki;Lee, Dong-Ryul
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.3
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    • pp.203-212
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    • 2008
  • Objective: During cryopreservation process, cold shock and cryo-injury affect the fertilizing capacity of the sperm by damaging cell membranes with loss of functional integrity. A longstanding concept for preventing the cryo-damage is to stabilize the plasma membrane by incorporating cholesterol. This study was to determine the effects of cholesterol in freezing media on the motility and functional integrity of human sperm after cryopreservation. Methods: Control group (non-cholesterol treated) and different concentrations of cholesterol-treated sperm (14 healthy males) were frozen and thawed. After freezing and thawing of sperm, the quality of sperm was evaluated by sperm analysis, acrosome reaction test and sperm chromatin structure assay. Results: When human sperm were incubated in sperm freezing medium (SFM) containing $0.5{\mu}g$ cholesterol and then freezing/thawing, the motility of sperm have significantly improved compared to those untreated cholesterol ($33.46{\pm}1.48%$ vs. $30.10{\pm}1.07%$, p<0.05). The rate of calcium ionophore-induced acrosome reactions in post-thawed sperm was significantly higher than that ($53.60{\pm}1.60%$ vs. $47.40{\pm}1.86%$, p<0.05) in SFM containing cholesterol. Sperm chromatin structure assay revealed that DNA damage to the sperm in the cholesterol-treated group was lower than that of non-treated group. Conclusion: These results suggest that increased cholesterol content of sperm plasma membrane by supplementation of cholesterol in SFM improves sperm motility, capacitation status, and DNA integrity. Therefore, addition of cholesterol into SFM could be a useful for protecting human sperm from cold shock and cryo-injury during cryopreservation.

Effects of Cytokinin and Auxin on Organ Formation in Leaf Scale Tissue of Allium Sativum L. (Cytokinin과 Auxin이 Allium Sativum L.의 인엽배양(鱗葉培養)에서 기관(器官)의 분화(分化)에 미치는 영향(影響))

  • Park, Kyeong Han;Lee, Yeong Bok
    • Korean Journal of Agricultural Science
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    • v.7 no.2
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    • pp.65-76
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    • 1980
  • In order to determine the effects of cytokinin and auxin on organ formation from tissue of garlic cloves, leaf blades and basal tissues contained meristem of garlic (Allium sativum L.) cloves harvested in 1979 (old cloves) and 1980 (new cloves) were explanted on a MS medium contained various levels of BA ($N^6$-benzyl amino purine), NAA (naphthalene acetic acid), and 2, 4-D (2, 4-dichlorophenoxyacetic acid). And some of the new cloves were explanted on a media contained BA and NAA after chilling treatment at $4^{\circ}C$ for 10, 20, 30 and 40 days. 1. In a culture of leaf blades of old cloves, shoots were differentiated on a medium supplemented with 2mg/l of BA and NAA. 2. Callus was grown as a quite straw-coloured globular mass on a medium contained 0.2 or 2mg/l 2.4-D. 3. As subcultures of globular calli, shoots and roots were differentiated on a medium contained 2mg/l BA and 0.5 or 1 mg/l NAA, whereas no shoots was shown on a conterol. 4. Shoots were differentiated in a culture of leaf blades of new cloves, but they were not in an old cloves in control, and better effect was shown on a medium contained 2mg/l BA and 1mg/l NAA. However shoots were no differentiated from leaf blades chilled at $4^{\circ}C$ for 30 or 40 days at the same condition. 5. Large numbers of adventitious shoots could be obtained from basal region of garlic cultured on a medium contained 1mg/l BA and 4mg/l NAA, or 2mg/l BA and 2mg/l NAA.

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A Study on Expressing 3D Animation by Visual Direction : focused on 〈 How to train your dragon 〉 (시각적 연출에 의한 3D 입체 애니메이션 표현 연구: 〈드래곤 길들이기〉를 중심으로)

  • Kim, Jung-Hyun
    • Cartoon and Animation Studies
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    • s.26
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    • pp.1-30
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    • 2012
  • The purpose of animation is to give interesting stories to an audience through motion. To achieve the purpose, over the past century since its inception, animation has adopted many kinds of technologies, and thus developed diverse narrative methods and visual expression techniques. In addition, with the advancement of expression techniques, all elements making up animation have gradually been systemized, and at the same time, have helped express the worlds beyond the reality. As a result, people have faced the era when an audience can watch everything imaginated by an animation director on a big screen. These days, more efforts have been made in order for the audience to feel much more than enjoy pictures moving in a frame. In other words, the purpose of the animation is changing from the passive viewing of animation to feeling and sensing stuffs through the animation. In the center of the changing process is 3D technology which gives new interesting to an audience. Sometime ago, a 3D animation movie was produced in Korea. But it did not bring out box-office profits, for it failed to give satisfaction to an audience who expected high perfection and beauty being able to be rivalled to those of international 3D animation movies. The failure is attributable to the fact that the domestic 3D animation production industry is merely in the early stage, and has not sufficient human resources, technology, and experiences in producing 3D animation films. Moreover, the problem is that most studies on 3D focus on the technologies related to reenactment, but that few studies on the images, which an audience directly faces, have been conducted. Under the domestic circumstance, the study on stereoscopic image screen of , a 3D stereoscopic animation film which was released in 2010 and has been seen as the best successful 3D stereoscopic animation, is worthwhile. Thus this thesis conducted theoretical consideration and case analysis focusing on the visual direction that creates the pictures to deliver abundant three dimensional effect so that it can be used as a basic data when producing high quality-domestic 3D animation and training professional labor forces. In the result, it was found that the 3D animation was not a new area, but the area which has been expanded and changed by applying the characteristics of 3D image based on the principles of the existing media aesthetics. This study might be helpful to establish the foundation of the theoretical studies necessary for producing 3D animation contents for realizing the sense of reality.

Numerical and Experimental Study on the Coal Reaction in an Entrained Flow Gasifier (습식분류층 석탄가스화기 수치해석 및 실험적 연구)

  • Kim, Hey-Suk;Choi, Seung-Hee;Hwang, Min-Jung;Song, Woo-Young;Shin, Mi-Soo;Jang, Dong-Soon;Yun, Sang-June;Choi, Young-Chan;Lee, Gae-Goo
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.2
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    • pp.165-174
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    • 2010
  • The numerical modeling of a coal gasification reaction occurring in an entrained flow coal gasifier is presented in this study. The purposes of this study are to develop a reliable evaluation method of coal gasifier not only for the basic design but also further system operation optimization using a CFD(Computational Fluid Dynamics) method. The coal gasification reaction consists of a series of reaction processes such as water evaporation, coal devolatilization, heterogeneous char reactions, and coal-off gaseous reaction in two-phase, turbulent and radiation participating media. Both numerical and experimental studies are made for the 1.0 ton/day entrained flow coal gasifier installed in the Korea Institute of Energy Research (KIER). The comprehensive computer program in this study is made basically using commercial CFD program by implementing several subroutines necessary for gasification process, which include Eddy-Breakup model together with the harmonic mean approach for turbulent reaction. Further Lagrangian approach in particle trajectory is adopted with the consideration of turbulent effect caused by the non-linearity of drag force, etc. The program developed is successfully evaluated against experimental data such as profiles of temperature and gaseous species concentration together with the cold gas efficiency. Further intensive investigation has been made in terms of the size distribution of pulverized coal particle, the slurry concentration, and the design parameters of gasifier. These parameters considered in this study are compared and evaluated each other through the calculated syngas production rate and cold gas efficiency, appearing to directly affect gasification performance. Considering the complexity of entrained coal gasification, even if the results of this study looks physically reasonable and consistent in parametric study, more efforts of elaborating modeling together with the systematic evaluation against experimental data are necessary for the development of an reliable design tool using CFD method.

Effect of Green Tea Extract on Cisplatin- or Doxorubicin-Induced Cytotoxicity in Human Lung Cancer Cell Lines (사람 폐암 세포주에서 시스플라틴이나 독소루비신의 세포독성에 미치는 녹차 추출물의 영향)

  • Lee, Byoung-Rai;Park, Jae-Yoon;Park, Pyoung-Sim
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.5
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    • pp.619-624
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    • 2011
  • Tea extract (TE) has been shown to have anti-tumor properties in a wide variety of experimental systems. We evaluated green tea extract (GTE) as a biochemical modulator for the antitumor activity of cisplatin and doxorubicin in the treatment of human lung cancer A549 cells. Cells were grown in RPMI-1640 medium supplemented with 10% (v/v) heat-inactivated fetal bovine serum and two antibiotics (100 units/mL penicillin and $100\;{\mu}g$/mL streptomycin). Two types of TE, epigallocatechin galate (EGCG) and GTE, were used in this experiment. The cells were seeded at $1{\times}10^4$ cells/well in the RPMI-1640 media with or without TE ($100\;{\mu}g$/mL) and then treated with different concentrations of doxorubicin ($0{\sim}14\;{\mu}g$/mL) or cisplatin ($0{\sim}35\;{\mu}g$/mL). After incubation in 5% $CO_2$ at $37^{\circ}C$ for 24 hr, cell viability was determined with a MTT assay. We used a Western blot to detect the influence of EGCG and GTE on the expression of p53 and caspase-3 genes in the A549 cells. A549 cell viability decreased to 15% with a $10\;{\mu}g$/mL concentration of cisplatin, and to 21% with a $8\;{\mu}g$/mL concentration of doxorubicin, as measured with the MTT assay. However, pre-treatment of the cells with EGCG ($100\;{\mu}g$/mL) or GTE ($100\;{\mu}g$/mL) resulted in decreased cell viability with $6\;{\mu}g$/mL of cisplatin and $4\;{\mu}g$/mL of doxorubicin. There was no apparent change in cell viability between EGCG or GTE administration in cisplatin- or doxorubicin-induced cytotoxicity in A549 cells. The levels of p53 and caspase-3 in the A549 cells increased with both EGCG and GTE treatment. We found that GTE could potentially affect cisplatin- or doxorubicin-induced cytotoxicity of A549 cells, which may be useful in the chemotreatment of cancer.

The Effects of Human Follicular Fluid on Embryonal Development of Mouse in In Vitro Culture (체외배양에서 인간 난포액이 생쥐의 배 발달에 미치는 영향)

  • Min, Bu-Kie;Choi, Ki-Wook;Kim, Kie-Suk;Lee, Hee-Sub;Hong, Ki-Yeon;Lee, Bong-Ju;Lee, Sun-Young;Park, Seung-Teak
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.171-178
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    • 1999
  • The follicular fluid (FF) of ovary contains various biological active products which affected on the growth of follicles and the fertilization of oocyte in physiological reproductive process of mammals. This study was designed to determine the effects of human FF on fertilization of oocyte and embryonal development in vitro culture. The FF was prepared as clear without blood contamination by needle aspiration from mature follicles of human at the time of oocytes retrieval for in vitro fertilization (IVF). As the medium for culture in vitro of embryonal cells, human tubal fluid (HTF) supplemented with follicular fluids at concentrations of 10%, 40% and pure FF were used. These effects were compared to control group of cultured embryos in HTF supplemented with 0.4% BSA (bovine serum albumin). For IVF, 64 eggs in control group, 67 eggs in 10% FF, 57 eggs in 40% FF and 64 eggs in pure FF were respectively allocated. And the rates of fertilization were almost similar in all groups as resulting 82.81% in control, 85.07% in 10% FF, 87.71% in 40% FF and 81.25% in pure FF. On the examination for embryonal cleavage from fertilized eggs, the rates of developing to 4 cell stage was similar in all groups, as results 98.11% in control, 98.27% in 10% FF and 98% in 40% FF but 78.84% in pure FF. And the rates of developing to 8-16 cell stage were significantly reduced as 44% in 40% FF and 44.23% in pure FF (p<0.05) compare to 71.69% in control media. As likewise, the rates of developing to morular stage were also significantly reduced to 36% (p<0.05) and 21.15% (p<0.01) respectively in 40% FF and pure FF. And the rates to blastocystic stage of embryo was lowest as 7.69% in pure FF (Table 1). The quality of embryonal cells on cleavage to the 8-16 cell stage was poorer, higher concentrations of FF. The rates of grade 1 in pure FF, as 23.07%, was lowest compare to those of other groups, in which the rates of grade 1 in control, 10% FF and 40% FF were 58.49%, 47.36% and 34% respectively. And on the contrary, the rate of grade 4 in pure FF was highest as 23.07%, while those were 5.66% in control, 8.77% in 10% FF and 20% in 40% FF (Table 2). On the viability of embryos, the rate of embryonal cell death was more rise, at the higher concentrations as well as longer exposure in the follicular fluid. At 48 hours after in vitro culture of embryos, the rate of survival embryos in pure FF was markedly lowered as 44.23%, compare to that of control (p<0.05). But there was not significant difference between the rates of survival embryos in each group beside the pure FF, which the rates were 77.35% in control, 70.17% in 10% FF and 60% in 40% FF respectively. And at 72 hours after in vitro culture, the rates of survival embryos were also significantly dropped to 21.15% in pure and 36% in 40% at concentration of FF compare to 62.26% in control (p<0.05, p<0.01). Finally, the rate of embryonal death at 96 hours after in vitro culture was highest as 82.69% in pure FF among all groups which those were 35.84 in control, 56.14% in 10% FF and 64% in 40% FF respectively (Fig. 1, 2, 3). In conclusion, this study suggests that the FF has no effects, in particular, to the in vitro fertilization of oocytes but exerted a bad effect to the cleavage, quality and viability of the embryonal cells during in vitro culture. However, the FF is harmful on embryonal development at conditions in higher concentration and especially on the embryos after $8{\sim}16$ cell stage.

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EFFECT OF NERVE GROWTH FACTOR GENE INJECTION ON THE NERVE REGENERATION IN RAT LINGUAL NERVE CRUSH-INJURY MODEL (백서 설신경 압박손상모델에서 신경성장인자 유전자 주입이 신경재생에 미치는 영향)

  • Gao, En-Feng;Chung, Hun-Jong;Ahn, Kang-Min;Kim, Soung-Min;Kim, Yun-Hee;Jahng, Jeong-Won;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.5
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    • pp.375-395
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    • 2006
  • Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

Effect of Different Nutrient Solution and Light Quality on Growth and Glucosinolate Contents of Watercress in Hydroponics (배양액의 종류 및 광질이 물냉이의 생육 및 Glucosinolate 함량에 미치는 영향)

  • Choi, Jae Yun;Kim, Sung Jin;Bok, Kwon Jeong;Lee, Kwang Ya;Park, Jong Seok
    • Journal of Bio-Environment Control
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    • v.27 no.4
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    • pp.371-380
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    • 2018
  • Aim of this study was to investigate the effects of different nutrient solutions and various light qualities generated by LED on the growth and glucosinolates contents of watercress (Nasturtium officinale) grown under hydroponics for 3 weeks. The seeds of watercress were sown on crushed rockwool media and raised them for two weeks. They were transplanted in a semi-DFT (deep flow technique) hydroponics system. A controlled-environment room was maintained at $20{\pm}1^{\circ}C$ and $16{\pm}1^{\circ}C$ temperatures and $65{\pm}10%$ and $75{\pm}10%$ relative humidity (day and night, respectively), with a provided photosynthetic photon flux density (PPFD) of $180{\pm}10{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ and a photoperiod of 16/8h. To find out the best kinds of nutrient solutions for growing watercress, Otsuka House 1A (OTS), Horticultural Experiment Station in Korea (HES), and Netherland's Proefstaion voor Bloemisterij en Gasgroente (PBG) were adapted with initial EC of $1.0-1.3dS{\cdot}m^{-1}$ and pH of 6.2, irradiating PPFD with fluorescent lamps (Ex-1). Either monochromatic (W10 and R10) or mixed LEDs (R5B1, R3B1, R2B1G1, and W2B1G1) were irradiated with a differing ratio of each LED's PPFD to understanding light quality on the growth and glucosinolates contents of watercress (Ex-2). Although significant difference in the shoot growth of watercress was not found among three nutrient solutions treatments, but the root fresh weight increased by 13.7% and 55.1% in PBG and OTS compared to HES, respectively. OTS increased the gluconasturtiin content by 96% and 65% compared to PBG and HES. Compared with the white light (W10), the red light (R10) showed a 101.3% increase in the shoot length of watercress. Increasing blue light portion positively affected plant growth. The content of total glucosinolates in watercress was increased by 144.5% and 70% per unit dry weight in R3B1 treatment compared with R2B1G1 and W10 treatments, respectively. The growth and total glucosinolates contents of the watercress were highest under R3B1 among six light qualities.

Effect of Cryopreservation Day on Pregnancy Outcomes in Frozen-thawed Blastocyst Transfer (동결 해동한 포배 이식에 있어서 동결시기가 임신결과에 미치는 영향)

  • Kim, Hyun-Jung;Kim, Chung-Hyon;Lee, Joong-Yeup;Kwon, Jae-Hee;Hwang, Do-Yeong;Kim, Ki-Chul
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.57-64
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    • 2010
  • Objectives: Likewise fresh cycle, it is also important to select right blastocysts for transfer in purpose of improving the pregnancy and implantation rates in frozen-thawed embryo transfer (ET) cycles. To investigate the relationship between the developmental velocity at the time of cryopreservation and pregnancy rates, we compared pregnancy rates between the day 5 cryopreservation group and the day 6 cryopreservation group. Methods: Transfers of frozen-thawed blastocysts which had been cryopreserved by vitrification on day 5 or day 6 were performed between January 2006 and June 2007. Ethylene glycol, DMSO, and pull and cut straws were used for vitrification and artificial shrinkage was done in expanded blastocysts. Thawing was performed on the day before transfer and thawed blastocysts were cultured in for 15~18 hrs in Quinn's blastocyct media. Blastocyst survival was assessed before transfer and post-thaw survival was defined as >50% of cells remaining intact and blastocoele re-expansion by the time of transfer. Results: Transfers of thawed blastocyst had been cryopreserved on day 5 were 52 cycles and 41 transfer cycles were cryopreserved on day 6. Patient characteristics, the number of transferred embryos and the survival rate of thawed blastocysts were not different in each cryopreservation day. But the biochemical pregnancy, clinical pregnancy, ongoing pregnancy, and implantation rate were significantly high in transfer of frozen-thawed blastocyst which were cryopreserved on day 5. Conclusions: The clinical pregnancy and implantation rate of day-5 blastocyst showed significantly higher than those of day-6 blastocyst in frozen-ET cycles. This result indicated that developmental rate of blastocyst at cryopreservation time in frozen-thawed cycle is discriminative marker of pregnancy outcome as like in fresh cycle.