• Title/Summary/Keyword: in vitro induction

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Alternating Acquisition Technique for Quantification of in vitro Hyperpolarized [1-13C] Pyruvate Metabolism

  • Yang, Seungwook;Lee, Joonsung;Joe, Eunhae;Lee, Hansol;Song, Ho-Taek;Kim, Dong-Hyun
    • Investigative Magnetic Resonance Imaging
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    • v.20 no.1
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    • pp.53-60
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    • 2016
  • Purpose: To develop a technique for quantifying the $^{13}C$-metabolites by performing frequency-selective hyperpolarized $^{13}C$ magnetic resonance spectroscopy (MRS) in vitro which combines simple spectrally-selective excitation with spectrally interleaved acquisition. Methods: Numerical simulations were performed with varying noise level and $K_p$ values to compare the quantification accuracies of the proposed and the conventional methods. For in vitro experiments, a spectrally-selective excitation scheme was enabled by narrow-band radiofrequency (RF) excitation pulse implemented into a free-induction decay chemical shift imaging (FIDCSI) sequence. Experiments with LDH / NADH enzyme mixture were performed to validate the effectiveness of the proposed acquisition method. Also, a modified two-site exchange model was formulated for metabolism kinetics quantification with the proposed method. Results: From the simulation results, significant increase of the lactate peak signal to noise ratio (PSNR) was observed. Also, the quantified $K_p$ value from the dynamic curves were more accurate in the case of the proposed acquisition method compared to the conventional non-selective excitation scheme. In vitro experiment results were in good agreement with the simulation results, also displaying increased PSNR for lactate. Fitting results using the modified two-site exchange model also showed expected results in agreement with the simulations. Conclusion: A method for accurate quantification of hyperpolarized pyruvate and the downstream product focused on in vitro experiment was described. By using a narrow-band RF excitation pulse with alternating acquisition, different resonances were selectively excited with a different flip angle for increased PSNR while the hyperpolarized magnetization of the substrate can be minimally perturbed with a low flip angle. Baseline signals from neighboring resonances can be effectively suppressed to accurately quantify the metabolism kinetics.

Induction of FSTL1 Gene in Rat Myometrium by Exogenous Estrogen

  • Kim Yoon-Sik;Kim Yeon;Bae Hyung-Joon
    • Biomedical Science Letters
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    • v.10 no.4
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    • pp.479-483
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    • 2004
  • The aim of this study was carried to the induction of follistatin-like 1 (FSTL1) in rat myometrium tissue by exogenous estrogen. PCR was performed by using estrogen treatment after ovariectomy of myometrium mRNA and only ovariectomy of myometrium mRNA and on ovariectomy of myometrium mRNA in rat normal uterus tissue. The PCR products were visualized on agarose gel (1.2%) electrophoresis. FSTL1 mRNA expression level was significantly higher (**P<0.001 or *P<0.05) in estrogen treatment after ovariectomy than in only ovariectomy, and was significantly higher (**P<0.001) in no ovariectomy than in estrogen with treatment after ovariectomy. In conclusion, Although the mechanisms of FSTL1 gene were uncertain, It also might be inducted by exogenous estrogen. And also it is thought to be related to the estorgen mechanism.

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Immunohistochemical analysis of effects of UVA exposure to the human fibroblasts in the skin equivalent model

  • Kazuhiro Shimizu;Fumihide Ogawa;Bae, Sang-Jae;Yoichiro Hamasaki;Ichiro Katayama
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.500-502
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    • 2002
  • In vitro and in vivo studies have reported the induction of matrix metaloproteinase (MMP)-1 in the fibroblasts by ultraviolet (UV) A irradiation. We constructed the skin equivalent model using HaCaT cells as keratinocytes and human neonatal dennal fibroblasts as fibroblasts in the present study. The induction of MMP-l in the fibroblasts was confirmed immunohistochemically 6 hours after UVA irradiation using this model. This model was simply composed of human keratinocytes and fibroblasts. To our knowledge, there have been a few papers concerning the skin equivalent model in the field of photobiology. The effect of UVA exposure to fibroblasts through keratinocytes was examined using this model. The cross-talk can be examined between keratinocytes and fibroblasts. This model can be a useful tool in the field of photobiology.

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Adventitious Shoots Regeneration from Seed Explants of Xanthoceras sorbifolium

  • Hyunseok Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.58-58
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    • 2020
  • Xanthoceras sorbifolium Bunge (yellowhorn) is a woody tree in the soapberry family, Sapindaceae, native to northern China. This species has been identified as a major woody bioenergy plant for bio-diesel production because of high oil content in seed. But the flowers do not bear fruit well while the many flowers blooming. This study was performed to regenerate in vitro plantlet using adventitious shoot formation. To establish the protocol of plant regeneration, adventitious shoots formation rate in the culture of cotyledon of immature zygotic embryos was 68.6% in 1/2 MS medium with 0.5 mg l-1 BA and 3% sucrose (w/v). In the culture of cotyledons of mature zygotic embryos, induction of adventitious shoots was needed to contain high sucrose in pre-culture medium and the frequency of shoot induction was 64.4%. Multiple shoots were induced in 0.5 mg l-1 TDZ, and rooting of shoot was induced 4.0 mg l-1 IBA. Flow cytometry analysis revealed that all the regenerated plantlets were diploid.

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Induction of G1 arrest and apoptosis mediated by a novel nucleoside analog, LJ-331 in human leukemia HL-60 cells

  • Lee, Eun-Jin;Shin, Dea-Hong;Jeong, Lak-Shin;Lee, Sang-Kook
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.86-86
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    • 2003
  • In a continuous effort to develop novel anticancer agents we newly synthesized and evaluated the antitumor activity of nucleoside analogues. One analogue, 4 - [2-Chlor-6- (3-iodo- benzy lamino) -purin -9-yl]- 2,3-dihydroxy-cyclopentanecarbo xylic acid methylamide (LJ-331), has been shown to exert a potent inhibition of human cancer cell growth in vitro including human lung (A549), stomach (SNU-638) and leukemia (HL-60) cancer cells. Following mechanism of action study revealed that LJ - 331induces cell cycle arrest at the G1 phase in HL-60 cells and evokes apoptotic phenomena such as an increase in DNA ladder intensity and chromatin condensation by a dose- and time-dependent manner. LJ-331 also activated the caspase-3 activity in HL-60. This result suggests that the growth inhibition of human cancer cells by LJ-331 might be related to the cell cycle arrest and induction of apoptosis.

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In Vivo Artificial Parthenogenetic Treatments on Live Silkworm Moth, Bombyx mori Can Induce Higher Parthenogenesis (살아있는 누에 나방(Bombyx mori)에 대한 인공적 단위 발생 처리의 단위 발생란 유발 촉진 효과)

  • Bae, Hee Eun;Lee, Yoon Kyung;Park, So Hyun;Lee, Seul-bi;Lee, Sang Mong
    • Journal of Life Science
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    • v.29 no.2
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    • pp.272-278
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    • 2019
  • The silkworm performs sexual reproduction for the production of its healthy offsprings from generations to generations. Parthenogenesis in the silkworm, Bombyx mori acquires immense use in the development of outstanding homozygouse lines with higher viability, hybrid vigour, combining ability and less phenotypic variability, and it can serve as a powerful tool in controlling sex of the offsprings as well as a useful tool in selection of breeding schemes. However, naturally occuring parthenogenesis in silkworm could not be found so far. Fortunately, artificial induction of parthenogenesis is possible in silkworm. So, it is very important to find out novel methods for induction of parthenogenesis. We investigated to attempt to get a novel parthenogenetic method. Accordingly, parthenogenetic studies on between unfertilized in vivo ovarian eggs of live silkworm moth(novel) and unfertilized in vitro ovarian eggs(conventional) taken out from live silkworm moth were investigated by hot water ($46^{\circ}C$), hot air ($46^{\circ}C$) and low temperature ($0^{\circ}C$ and $-20^{\circ}C$) treatments. The best ratio of parthenogenetic eggs was obtained with in vivo ovarian eggs of live silkworm moth rather than with in vitro ovarian eggs taken out from live silkworm moth in all the treatments. The optimum exposure time absolutely depended upon the temperatures of treatments and the forms of in vivo or in vitro ovarian eggs. From these results, we expect that in vivo artificial parthenogenetic treatments on live silkworm moth will be useful for the higher induction of parthenogenesis in the silkworm, B. mori.

Mass Production of Adventitious Roots of Eleutherococcus sessiliflorus through the Bioreactor Culture

  • Seo Jin-Wook;Shin Cha-Gyun;Choi Yong-Eui
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.187-191
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    • 2003
  • This paper reported the establishment of mass production system of adventitious roots of Eleutherococcus sessiliflorus through the shake flask and bio-reactor culture. Induction of adventitious roots was started from the explants of germinated somatic embryos on half-strength Murashing and Skoog (MS) solid medium. The frequency of adventitious root formation was better in the explants comprising the basal hypocotyl parts than root explants alone. Among the different auxins tested (NAA, IBA and IAA), frequency of adventitious root induction was highest on medium with 0.5 mg/L NAA, and produced $16.3\pm1.9$ roots per explant. In shake-flask culture, deletion of $NH_4NO_3$ of MS medium was effective for induction of adventitious root compared with both full and half-strength MS media. Fresh weight increase of induced adventitious roots was performed well in medium with 0.5 mg/L IBA. When adventitious roots produced in shake-flask culture were transferred to 10-liter bioreactor, 5.5 times of fresh weight increase was gained after one month of culture. HPLC analysis revealed that the amount of eleutheroside E and E1 was higher in in vitro cultured adventitious roots than the 3 year-old field cultivated root barks of Eleutherococcus sessiliflorus. The content of eltutheroside B was much lower in adventitious roots than that of field cultivated one.

Micropropagation of a Rare Species, Abeliophyllum distichum Nakai. via axillary bud culture (액아배양에 의한 희귀 수종 미선나무의 기내번식)

  • 문흥규;석진영;권영진;손성호
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.133-136
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    • 1999
  • Different kinds of cytokinins and auxins were tested for both shoot induction and rooting in a rare species, Abeliophyllum distichum Nakai. BA in WPM medium was the most effective in shoot induction, whereas zeatin seemed to be the most suitable for shoot elongation. Kinetin, at the concentration of 2.0~5.0 mg/L showed an effect in shoot induction, but the effect was inferior to BA and zeatin. Rapid shoot elongation could be achieved when the cultures were maintained on the diffuse light condition (below 500 lux) regardless of cytokinin treatments. For in vitro rooting, IBA was investigated as the best type of auxin tested when half strength GD medium was incorporated. The frequency of rooting using the plant growth regulator and medium just mentioned above was revealed as approximately 90%. In addition, the survival rate of rooted plantlets was almost 100% in an artificial soil mixture.

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Studies on Effects of the Administration of $PGF_{2\alpha}$ GTH and Antibiotics in Repeat Breeders (Prostaglandin $F_{2\alpha}$, GTH 및 Antibiotics의 투여가 Repeat breeder의 치료에 미치는 영향)

  • 정병현
    • Korean Journal of Animal Reproduction
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    • v.9 no.1
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    • pp.66-71
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    • 1985
  • After the induction of estrus by the PGFF2$\alpha$-GTH combination on the 118 repeat breeder cow during of 6-13 days of the estrus cycle, the effects of GTH, antibiotics, antibiotics-GTH combination were compared. The results of this study, the following conclusion were obtained: 1. The rate of estrus was reduced the administration of GTH before the administration of PGF2$\alpha$. 2. The rate of ovulation delayed was 24.6% in the group with the administration GTH, but 34.1% without the administration GTH. Duration of ovulation was shortened by GTH. 3. The impregnation induction by the administration of PGF2$\alpha$, PGF2$\alpha$-antibiotics, PGF2$\alpha$-GTH-antibiotics were 41.7%, 69.5%, 55.6% and 80.8% respectively injection in the uterus was more effective than the administration. Before or after the A.I., the above combination was most effective. 4. In antibiotics sensitivity test in vitro of the intravagina bacteria, 5.0%, 12.7% and 60.0% of bacteria were sensitive to Penicilline, Streptomycin and Gentamicin respectively.

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Plant Regeneration from Turnip (Brassica rapa ssp. rapifera) Organs

  • Gendaram Sarantuya;Bae Chang-Hyu
    • Plant Resources
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    • v.8 no.3
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    • pp.286-292
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    • 2005
  • Shoot induction system was developed in the recalcitrant plant species, Brassica rapa ssp. rapifera by using optimum selection of profit organ, phytohormone combination, seedling age and kind of culture container. Out of in vitro cultured leaf segment, petiole, hypocotyl, and cotyledon with petiole, only cotyledon with petiole derived from 4 day-old seedlings induced multiple shoot. The optimum combination of auxin and cytokinin for the multiple shoot induction was MS medium containing 5mg/L BA and 0.5mg/L NAA. The major factors for multiple shoot propagation were part of plant organ, age of seedling, and ratio of auxin and cytokinin. In addition, shoot regeneration was promoted in the 100ml Erlenmeyer flask compared with the $90mm{\times}20mm$ Petri-dish. The induced shoots formed roots easy on MS medium containing 0.1mg/L IBA and the whole plants were successfully cultivated in soil.

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