• Title/Summary/Keyword: survival and in vitro development

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Establishment of high frequency plant regeneration system from leaf explants of Pinellia koreana via bulblets formation

  • Oh, Myung-Jin;Park, Jong-Mi;Lee, Bu-Youn;Choi, Pil-Son;Tae, Kyoung-Hwan;Kim, Suk-Weon
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.193-196
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    • 2009
  • Pinellia koreana K-H Tae & J-H Kim is a recently discovered Korea endemic medicinal plant species whose natural habitat is rapidly destroyed by industrial development. Described in this paper are culture conditions for high frequency plant regeneration via bulblet formation from leaf explant cultures of P. koreana. Leaf explants formed white nodular structures and off-white calluses at a frequency of 91.2% when cultured on MS medium supplemented with 2 mg/L BA and 0.5 mg/L NAA. However, the frequency of white nodular structures and off-white calluses formation was slightly decreased with an increasing concentration of NAA up to 4 mg/L, where the frequency reached 31.7%. Most petiole explants did not form white nodular structures and off-white calluses except the combination treatment of 2 mg/L BA and 2 mg/L NAA. Upon transfer onto MS basal medium, over 90% of nodular structures gave rise to numerous bulblets and developed into plantlets. Plantlets regenerated from bulblets were transplanted to potting soil and grown to maturity at a survival rate of over 95% in a growth chamber. Therefore, the in vitro plant regeneration system of P. koreana obtained in this study will be useful for mass propagation and long-term preservation of genetic resources of P. koreana.

Antioxidant Favors the Developmental Competence of Porcine Parthenogenotes by Reducing Reactive Oxygen Species

  • Hossein, Mohammad Shamim;Kim, Yeun Wook;Park, Seon Mi;Koo, Ok Jae;Hashem, Md Abul;Bhandari, Dilip P;Jeong, Yeon Woo;Kim, Sue;Kim, Ji Hye;Lee, Eu Gine;Park, Sun Woo;Kang, Sung Keun;Lee, Byeong Chun;Hwang, Woo Suk
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.3
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    • pp.334-339
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    • 2007
  • Reactive oxygen species (ROS) generate during electrical activation of oocytes which has detrimental effects on embryo survival when overwhelmed. The present study was designed to investigate the ability of L-ascorbic acid, a novel water soluble antioxidant, to reduce the ROS level in developing embryos and their subsequent effects on embryo development in vitro. The compact cumulus oocyte complexes (COCs) were cultured in tissue culture medium (TCM)-199 supplemented with 10 ng/ml epidermal growth factor, 4 IU/ml pregnant mare serum gonadotropin (PMSG), and human chorionic gonadotropin (hCG) and 10% (v/v) porcine follicular fluid (pFF) for 44 h. After maturation culture, the denuded oocytes were activated with a single DC pulse of 2.0 kV/cm in 0.3 M mannitol solution containing 0.5 mM of HEPES, 0.1 mM of $CaCl_2$ and 0.1 mM of $MgCl_2$ for $30{\mu}s$ using a BTX Electro-cell Manipulator. The activated oocytes were cultured in modified North Carolina State University-23 (mNSCU-23) medium for 168 h. The level of $H_2O_2$ in each embryo was measured by the dichlorohydrofluorescein diacetate (DCHFDA) method at 48 h after activation. The blastocyst formation rate was significantly higher when culture medium was supplemented with 50 and $100{\mu}M$ L-ascorbic acid (31.2 and 38.7%, respectively) compared to non-supplemented (16.1%) group. Accordingly, significantly more cells in blastocyst were found for 50 and $100{\mu}M$ L-ascorbic acid (50.0 and 56.4, respectively) compared to 0 and $200{\mu}M$ L-ascorbic acid (36.5 and 39.8, respectively). L-ascorbic acid reduces the $H_2O_2$ level in developing embryos in a dose-dependant manner. The $H_2O_2$ level (pixels/ embryos) was 191.5, 141.0, 124.0 and 163.3 for 0, 50, 100 and $200{\mu}M$ L-ascorbic acid, respectively. So, we recommend to supplement 50 or $100{\mu}M$ L-ascorbic acid in porcine in vitro culture medium.

Blood Flow and Pressure Evaluation for a Pulsatile Conduit-Shaped Ventricular Assist Device with Structural Characteristic of Conduit Shape (관형의 구조적 특징을 갖춘 박동형 관형 심실보조장치의 혈류, 혈압 평가)

  • Kang, Seong-Min;Choi, Seong-Wook
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.35 no.11
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    • pp.1191-1198
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    • 2011
  • The use of a ventricular assist device (VAD) can raise the one-year survival rate without cardiac transplantation from 25% to 52%. However, malfunction of the VAD system causes 6% of VAD patients' deaths, which could possibly be avoided through the development of new VADs in which VAD malfunctions do not affect the patient's heart movement or hemodynamic state. A conventional VAD has an impeller or vane for propelling blood that can allow blood to regurgitate when the propelling force is weaker than the aortic pressure. In this paper, we developed a new pulsatile conduit-shaped VAD that has two valves. This device removes the possibility of blood regurgitation and has a small stationary area even when the pumping force is extremely weak. We estimated the characteristics of the device by measuring the outflow and the pressure of the pump in in-vitro and in-vivo experiments.

Development of S-Methylmethionine Sulfonium Derivatives and Their Skin-Protective Effect against Ultraviolet Exposure

  • Kim, Won-Serk;Kim, Wang-Kyun;Choi, Nahyun;Suh, Wonhee;Lee, Jinu;Kim, Dae-Duk;Kim, Ikyon;Sung, Jong-Hyuk
    • Biomolecules & Therapeutics
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    • v.26 no.3
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    • pp.306-312
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    • 2018
  • In a previous study, we have demonstrated that S-methylmethionine sulfonium (SMMS) confers wound-healing and photoprotective effects on the skin, suggesting that SMMS can be used as a cosmetic raw material. However, it has an unpleasant odor. Therefore, in the present study, we synthesized odor-free SMMS derivatives by eliminating dimethyl sulfide, which is the cause of the unpleasant odor and identified two derivatives that exhibited skin-protective effects: one derivative comprised (2S,4S)- and (2R,4S)-2-phenylthiazolidine-4-carboxylic acid and the other comprised (2S,4R)-, (2S,4S)-, (2R,4R)-, and (2R,4S)-2-phenyl-1,3-thiazinane-4-carboxylic acid. We performed in vitro proliferation assays using human dermal fibroblasts (hDFs) and an immortalized human keratinocyte cell line (HaCaT). The two SMMS derivatives were shown to increase hDF and HaCaT cell proliferation as well as improve their survival by protecting against ultraviolet exposure. Moreover, the derivatives regulated the expression of collagen type I and MMP mRNAs against ultraviolet exposure in hDFs, suggesting that these derivatives can be developed as cosmetic raw materials.

Ethanol extract of Aster glehni exhibits anti-inflammatory and anti-oxidant effects in RAW 264.7 cells and Caenorhabditis elegans (섬쑥부쟁이 에탄올 추출물이 대식세포와 예쁜꼬마선충에서의 항염증 및 항산화 효과 )

  • Mi-Kyung Seo;Han-Na Chu;Da-Bin Lee;Haeng-Ran Kim;In-Seon Hwang;Yong-Jin Jeong;Sung-Ran Yoon;Seok-Seong Kang;Kyeong-A Jang;Min-Sook Kang
    • Food Science and Preservation
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    • v.30 no.6
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    • pp.1095-1106
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    • 2023
  • This study investigated the anti-oxidative and anti-inflammatory effects of Aster glehni (AG) extract in RAW 264.7 cells and Caenorhabditis elegans. The total polyphenol and flavonoid contents were higher in the ethanol extracts than in the hot water extracts. As a result of measuring the moisture contents (%) and extraction yields (%) of AG and drying A. glehni for processing (DAG), 70% ethanol, which has the highest percentage of extraction yield, was selected as the final solvent. DPPH radical scavenging activity showed higher antioxidant activity of ethanol extracts of DAG than AG. The cytotoxicity assay of the AG or DAG ethanol extracts was treated at different concentrations (25, 50, and 100 ㎍/mL), and cell viability rates were higher than 80% at all concentrations. The LPS-stimulated nitric oxide (NO) production in RAW 264.7 was significantly reduced at all concentrations of AG and DAG groups. As a result of measuring the gene expression of iNOS, which induces NO production, the AG or DAG group decreased by 33% and 32%, compared with the phosphate buffer saline (PBS) group. Under inflammatory stress conditions, the survival rate of C. elegans treated with AG or DAG ethanol extract with LPS showed concentration-dependent improvement in survival rate compared with the PBS group. Considering these results, AG could potentially be developed as an antioxidant and anti-inflammatory functional food material.

Elimination of Apple stem grooving virus from 'Mansoo' pear (Pyrus pyrifolia L.) by an antiviral agent combined with shoot tip culture (항바이러스제 처리와 경정배양에 의한 배(Pyrus pyrifolia L.) '만수'의 Apple stem grooving virus 무병화)

  • Cho, Kang Hee;Shin, Juhee;Kim, Dae-Hyun;Park, Seo Jun;Kim, Se Hee;Chun, Jae An;Kim, Mi Young;Han, Jeom Hwa;Lee, Han Chan
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.391-396
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    • 2016
  • In this study, in vitro-cultured 'Mansoo' pear (Pyrus pyrifolia L.) plants infected with Apple stem grooving virus (ASGV) were used for testing the efficiency of the virus elimination methods. The shoot tips cut from infected plants were treated by thermotherapy ($37^{\circ}C$), cold therapy ($4^{\circ}C$), chemotherapy with ribavirin, and combination of these methods. Treatment periods were 2, 4, and 8 weeks, and concentrations of ribavirin were 20 and $40mg{\cdot}L^{-1}$. The efficiency of ASGV elimination was evaluated by reverse transcription polymerase chain reaction. The shoot survival rate was the highest at 100% after cold therapy, chemotherapy, and combination of two methods, while the rate was the lowest at 33.3% after thermotherapy for 2 weeks. The shoot survival rate after chemotherapy decreased gradually as the treatment period was prolonged. The ASGV elimination rate was the highest at 100% after ribavirin treatment at a concentration of $40mg{\cdot}L^{-1}$ and combination of ribavirin treatment and thermotherapy for 2 weeks, whereas the ASGV elimination rate after cold therapy was the lowest at 16.7%. However, the efficiency of ASGV elimination was enhanced up to 43.3% by the combination of cold therapy and ribavirin treatment. The efficiency of ASGV elimination for all treatments was increased as the treatment period was prolonged. Based on these results, we suggest that ribavirin treatment at a concentration of $20mg{\cdot}L^{-1}$ for 4 weeks or at a concentration of $40mg{\cdot}L^{-1}$ for 2 weeks combined with shoot tip culture was efficient for the elimination of ASGV from pear.

Antitumor compounds from fruiting bodies of cultivated fungus of Paecilomyces japonica

  • Nama Ryu;Lim, Soon-Sung;Jung, Sang-Hoon;Cho, Sae-Yun;Shin, Kuk-Hyun
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.185-185
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    • 1998
  • As part of our continuing attempts to evaluate biologically active compounds from fruiting bodies of cultivated fungus of Paecilomyces japonicus Yasuda, we conducted series of experiments on various fractions and compounds isolated by systematic fractionations. Our main efforts were concentrated on searching for compounds showing antitumor activities, which were tested on mice carrying Sarcoma-180 ascitic tumor. The antitumor activity was assessed by the life spans after these mice were administered Lp. with test compounds for consecutive 20 days. One of two pure compounds, which we have isolated to date, demonstrated significant prolongation of life span. ( Mean Survival Time: 30.3 days compared to that of control: 23.6 days). Structural analysis showed that this compound corresponds to D-mannitol. On the other hand, Ergosterol, another isolated pure compound didn't show efficient antitumor activity. We also obtained water-soluble fractions containing protein-bound polysaccharides and n-butantol fractions, which showed strong antitumor activities, 35.4(150%) and 32.1(136.0%) days of MST, respectively. In SRB assay, however, the test materials didn't show any toxic effects, but the level of acid phosphatase increased significantly when they were applied in cultured macrophage in vitro. Therefore, we concluded that antitumour activities might be attributed to immunostimulating rather than cytotoxic effects. Further experiments are underway to purify and structurally characterize new antitumour compounds from the active fractions.

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Effects of Cooling Rate and Equilibration Time on the Survival and Development of Frozen-thawed Bovine Immature Oocytes (동결속도와 평형시간이 소 미성숙 난자의 동결 융해후 생존율에 미치는 영향)

  • 양병철;양보석;성환후;임기순;최선호;장원경;진동일;임경순
    • Korean Journal of Animal Reproduction
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    • v.25 no.1
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    • pp.19-28
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    • 2001
  • The present study was undertaken to investigate the effects of cooling rate and equilibration time on the survival, in vitro maturation and development to embryos of frozen-thawed bovine immature oocytes(Germinal Vesicle Stage). The cryoprotectants are used 10% ethylene glycol(EG) as permeating cryoprotectant and 0.05M soc.ose(S) or trehalose(T) as low molecular weight nonpermeating cryoprotectants and 5% ficoll(F) or polyvinylpyrrolidone(PVP) as high molecular weight nonpermeating cryoprotectants. Four freezing solution were uysed in this experiment(EFT: 10% EG + 5% F + 0.05M T, EFS: 10% EG + 5% F + 0.05M S, EPT: 10% EG + 5% P + 0.05M T, EPS: 10% EG + 5% P + 0.05M S). The best equilibration time and freezing solution was 15 min in EPT(83% survival rate of frozen-thawed bovine immature oocytes). When frozen-thawed bovine oocytes were cultured following IVM and IVF, there was no significant difference in cleavage and development rates among the EFT, EFS, EPT and EPS solutions. When 9 blastocysts derived from frozen bovine oocytes were transferred to 6 recipients, two recipients were pregnant. And one was aborted at 45 days of pregnancy and the other had a stillbirth.

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Multidrug Resistance and Cytotoxicity of Anticancer Drug by Verapamil in Cisplatin Resistant Human Stomach Cancer Cell (Cispatin 내성인 사람 위암 세포주 SNU-1의 복합약제내성 및 Verapamil의 효과)

  • Son, Seong-Kweon;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.9 no.1
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    • pp.75-89
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    • 1992
  • The development of multi drug-resistant tumor cell population is a major problem in the chemotherapy of human cancer. These cells are often cross resistant to unrelated drugs and the precise mechanisms of multidrug resistant phenotype of tumor cells has not been fully elucidated. Cisplatin resistant tumor cell(SNU-1/$Cis_5$) was induced from human stomach cancer cell line(SNU-1) in vitro. Growth profiles of survival cells were observed during 5 days by thiazolyl blue (MTT) assay. To investigate the cross resistance of various anticancer drugs in SNU-1 and SNU-1/$Cis_5$, We compared the value of $IC_{50}$ - drug concentration at 50% survival of control and gained relative resistances (RR). The RR for SNU-1/$Cis_5$ were as follows; vinblastine, > 43.0 ; epirubicin, 22.9 ; dactinomycin, 16.0 ; etoposide, 15.0 ; vincristine, 9.2 ; adriamycin, 5.7 ; aclarubicin, 5.3. But 5-fluorouracil, methotrexate, daunorubicin have not cross resistance with cisplatin. Resistant inhibition values of $10{\mu}M$ verapamil for SNU-1/$Cis_5$ were as follows; vincristine, 13.1 ; epirubicin, 10.0 ; etoposide, 6.3 ; vinblastine, 4.4 ; dactinomycin, 3.6 ; daunorubicin, 2.4. Membrane proteins of 51,400 and 81,300 daltons were identified by radioiodination with SDS-PAGE, which might represented the drug resistance.

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Systematic Propagation of High Quality Garlic (Allium sativum L.) Through Shoot Apical Meristem Culture 1. Organogenesis from in Vitro Cultured Shoot-tips (생장점배양에 의한 우량마늘 체계적 증식 1.생장점배 양으로부터 기관형성)

  • Lee, Eun-Mo;Lee, Young-Bok
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.161-166
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    • 1994
  • Since garlics (Allium sativum L.) are propagated through cloves, infection by virus or other pathogens may become severe problem if not using high quality seed bulbs every year resulting in the reduction of yield and bulb quality, In order to solve this problem, the establishment of virus-free bulb production and its supply system have been required because no chemicals were found to eliminate viruses from seed bulbs. This experiment was conducted to develop an effective production technique of high quality seed bulbs using shoot-tip culture. Over 90% of shoot-tips explanted on January L 1990 were survived at constant temperature of either 20, 24 or 28$^{\circ}C$, wheres 88% at alternate temperature (28/20$^{\circ}C$). The growth of shoot and root was most vigorous at constant 24$^{\circ}C$, and least at alternate temperature (28/20$^{\circ}C$) condition. When shoot-tips were explanted June 21 to August 1,1991, survival and growth of shoot-tips was most vigorous on MS medium supplymented with 0.1 mg/L NAA and 2 mg/L kinetin and least 1 mg/L Gh$_3$. The shoot-tips taken from the seed bulbs stored at 4$^{\circ}C$ for 15 to 60 days were placed on MS medium, shoot growth and in vitro bulblet formation increased slightly as affected by the increase of told treatment period at 4$^{\circ}C$.

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