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Effect of Plant Growth Regulators on Plant Regeneration Through Somatic Embryogenesis of Medicago sativa L.

  • Kim, Young-Sook;Kim, Mi-Young;Yang, Moon-Sik
    • Journal of Plant Biotechnology
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    • v.6 no.2
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    • pp.87-90
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    • 2004
  • An efficient plant regeneration system in alfalfa (Medicago sativa L.) through somatic embryogenesis was established. Embryogenic callus was obtained by culture of hypocotyl segments on MS medium with 0.02mg $L^{-1}$ IAA and 1.0mg $L^{-1}$ zeatin after 45 days of culture. Embryogenic calli were converted to the somatic embryos when transferred to either MS medium without plant growth regulators (PGRs) or MS medium containing various cytokinin (BA, kinetin and zeatin). Most of the somatic embryos were developed into plantlets on MS medium supplemented with 0.1 mg $L^{-1}$ kinetin. Also, secondary embryos appeared on the surface of primary embryo but they showed abnormal growth. Regenerated plantlets were transplanted to pots containing vermiculite and perlite for further analysis.

Headspace-GC-MS에 의한 토양 중 MTBE와 BTEX의 동시분석법에 관한 연구

  • Sin Ho-Sang;An Hye-Sin;Ryu Sang-Hui;Kim Tae-Seung
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2005.04a
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    • pp.31-35
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    • 2005
  • 본 연구는 토양에 휘발유 첨가제인 MTBE와 휘발유의 주성분인 BTEX를 headspace 법에 의해 동시 분석하는 방법이다. 인산으로 pH를 2로 조절한 후 NaCl로 포화시킨 용액 5ml를 헤드스페이스 바이알에 보존제로 넣은 후 토양시료 약 2g을 이 용액에 침지시켜 시료 채취한 다음 헤드스페이스 장치에 넣고 $80^{\circ}C$에서 40분 가온하여 상부 기상의 일정량을 취해 GC-MS (SIM)으로 분석하였다. 본 분석법에 의한 검출한계는 methyl-tert-butyl ether(MTBE)와 benzene, toluene, ethylbenzene, o,m,p-xylene(BTEX)이 각각 0.1, 0.1, 0.1, 0.2, 0.1, 0.2 ng/g이었고, 직선성은 0.995이상이었으며, 재현성도 10%내외의 정밀한 값을 보였다. 실제 시료를 분석한 결과, MTBE가 3-6,993 ng/g의 농도분포를 보였고 total BTEX는 1 ng/g으로 검출되었다. 이 방법은 빠르고 정밀 정확한 분석법으로 공정시험법으로 활용가치가 높다.

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Multiple shoot induction and callus regeneration in Sarcostemma brevistigma Wight & Arnott, a rare medicinal plant

  • Thomas, T. Dennis;Shankar, Surabhi
    • Plant Biotechnology Reports
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    • v.3 no.1
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    • pp.67-74
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    • 2009
  • An efficient micropropagation protocol based on multiple shoot induction and callus regeneration has been standardized in Sarcostemma brevistigma, a rare medicinal plant. The nodal cuttings were cultured on MS medium supplemented with BA ($0.5-8{\mu}M$) or Kn ($0.5-8{\mu}M$) alone or in combination with NAA ($0.5-1.5{\mu}M$). Maximum multiple shoot induction was observed on MS medium supplemented with $4{\mu}M$ BA. On this medium, 100% cultures responded with an average number of 11.3 shoots per explant. However, the average shoot length was limited to only 0.9 cm on this medium. The addition of $1{\mu}M$ NAA along with $4{\mu}M$ BA gave rise to an average number of 10.9 shoots with an average shoot length of 1.8 cm. Luxuriantly growing callus was obtained on MS medium supplemented with BA ($5{\mu}M$) and 2,4-D ($2{\mu}M$). The callus was subcultured on MS medium supplemented with BA ($2-15{\mu}M$) or Kn ($2-15{\mu}M$) alone or in combination with NAA ($0.5-2{\mu}M$) for shoot organogenesis. Optimum callus regeneration was obtained on MS medium supplemented with $10{\mu}M$ BA and $1{\mu}M$ NAA. On this medium, 100% cultures responded with an average number of 13.4 shoots per culture. The shoots obtained via multiple shoot induction and organogenesis were rooted on half-strength MS medium supplemented with NAA ($1-7{\mu}M$) or IBA ($1-7{\mu}M$). IBA was better than NAA in terms of both the percentage of cultures that responded and the average number of roots per explant. The rooted shoots were successfully transplanted to soil with 86% success. This standardized protocol will help to conserve this rare medicinal plant.

Effect of Forsythia Viridissima Extracts on Antioxidative System and Lipid Peroxidation of Liver in Rats Fed High-Cholesterol Diet (연교 (Forsythia Viridissima Lindl.) 추출물이 고콜레스테롤 식이 흰쥐 간조직의 항산화계와 지질과산화에 미치는 영향)

  • 이정민;최상원;조성희;이순재
    • Journal of Nutrition and Health
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    • v.36 no.10
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    • pp.990-996
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    • 2003
  • This study was conducted to investigate the effects of Forsythia viridissima Lindl. (FVL) on antioxidative defense system and lipid peroxidation of liver in rats fed high-cholesterol diet. Sprague-Dawley male rats weighing 100 $\pm$ 10 g were randomly assigned into five experimental groups fed 0.5% cholesterol ; HC group which was not supplemented FVL extract, 0.05% methanol extract diet group (MSI group), 0.1% methanol extract diet group (MS2 group), 0.025% ethylacetate-souble fraction diet group (ES1 group) and 0.05% ethylacetate-souble fraction diet group (ES2 group). Experimental diets were fed ad libitum to the rats for 3 weeks. The hepatic xanthine oxidase (XOD) activity in the MS2 group was decreased to 20% as compared to HC group. The activities of hepatic superoxide dismutase (SOD) and catalase (CAT) were not significantly different among all the high cholesterol diet groups. The hepatic glutathione peroxidase (GSHpx) activity in MS2, ES2 groups were significantly increased as compared to HC group. The hepatic glutathione S-transferase (GST) activity in the MS2 group was increased to 20% as compared to HC group. The levels of hepatic TBARS in the MS1, MS2, ES1 and ES2 groups were reduced by 13%, 21%, 13% and 21%, respectively, as compared with HC group. The contents of lipofuscin in liver tissue was not significantly different among all the experimental groups. The results indicate that FVL extract may reduce oxidative damage by activating antioxidative defense system of liver in rats fed high-cholesterol diets.

In vitro plant regeneration from axillary buds of Hibiscus syriacus L.

  • Jeon, Seo-Bum;Kang, Seung-Won;Kim, Wan-Soon;Lee, Gung-Pyo;Kim, Sun-Hyung;Seo, Sang-Gyu
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.174-178
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    • 2009
  • Presently, we report a simple, reproducible and high frequency plant regeneration in Hibiscus syriacus L. using axillary buds. H. syriacus was regenerated from axillary buds directly or through a callus phase. Regenerated shoots were directly induced from young and fresh axillary buds cultured on Murashige and Skoog medium (MS) supplemented with 0.01 mg/L of the growth regulator thidiazuron (TDZ) after 2 weeks of culture. Directly induced shoots were transferred to hormone-free MS medium and root development was observed after 6 weeks. On the other hand, old and stale axillary buds were regenerated to shoots via callus induction on MS medium containing 0.01–2 mg/L TDZ after 4 weeks. A TDZ concentration of 0.01 mg/L was most effective in callus formation. Green callus was transferred to MS medium containing 0.01 mg/L α-naphthalene acetic acid (NAA) and 0.5 mg/L benzylaminopurine (BA). After 4 weeks, callus had developed into multiple shoots. Plantlets were formed from 10 week cultures of single shoots on hormone-free MS medium. Regenerated plantlets were cultured on MS medium for one month and then transferred to pots containing garden soil. Potted plants were acclimatized for one month and grown to maturity under greenhouse conditions. The present study has shown that various concentrations of plant growth regulator can be effective for in vitro plant regeneration of H. syriacus. The direct and indirect regeneration protocol presented here will be useful for understanding the manipulation and propagation of H. syriacus.

NUTRITIONAL QUALITY OF WHOLE CROP CORN FORAGE ENSILED WITH CAGE LAYER MANURE. I. QUALITY, VOLUNTARY FEED INTAKE AND DIGESTIBILITY OF THE SILAGES IN GOATS

  • Kim, J.H.;Yokota, H.;Ko, Y.D.;Okajima, T.;Ohshima, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.1
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    • pp.45-51
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    • 1993
  • With the purpose to utilize cage layer manure (CLM), whole crop corn forage was ensiled with 30% CLM (MS silage) and without CLM (CS silage). MS silage was significantly (p < 0.05) higher in pH value, total VFA, propionic and butyric acids, and the ratio of ammonia nitrogen to total nitrogen, but lower (p < 0.05) in lactic acid and water soluble carbohydrate (WSC) contents than CS silage. Digestibility was evaluated using Shiba strain Japanese goats. Urea was supplemented to CS silage at feeding to adjust nitrogen intake to MS silage (US silage). There were no differences in digestibilities of dry matter (DM) and energy among the three silages. However, US silage showed higher (p < 0.05) digestibility of crude protein, but digestibilities of NDF, ADF, hemicellulose and cellulose were higher (p < 0.05) in MS silage. Nitrogen retention was positive in US and MS silages, but it was negative in CS silage. Voluntary feed intake of goats was 11.02, 12.03 and 13.34 g of DM per metabolic body weight ($kg^{0.75}$) for 10 minutes, for CS, US and MS silages, respectively.

Quantitative measurement of salivary testosterone in Korean adults by stable isotope-dilution liquid chromatographyelectrospray-tandem mass spectrometry

  • Lee, Sang-Hoo;Kwon, Soon-Ho;Shin, Hye-Jin;Park, Ji-Myeong;Lim, Hwan-Sub;Lee, Kyoung-Ryul;Kim, Young-Jin
    • BMB Reports
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    • v.43 no.11
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    • pp.761-765
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    • 2010
  • Salivary testosterone levels in Korean adults were quantitatively measured for the first time by liquid chromatography-electrospray-tandem mass spectrometry (LC ESI MS/MS). Salivary testosterone was separated on a multiple reaction monitoring (MRM) chromatogram within 7 min. The LC ESI MS/MS assay was validated over the linearity range of 0.01-2.00 ng/ml (r=0.99987) using testosterone-$d_3$ as an internal standard. The lower limit of quantification (LOQ) was 0.01 ng/ml. The intra- and inter-assay precisions were 1.54% to 4.09% and 0.96% to 4.29%, respectively. The mean recovery was 93.32% (range 88.43-98.05%). The validated assay was then applied to measure the salivary testosterone levels of Korean adults. In men, the salivary testosterone level collected between 9:00-11:00 am was approximately 2.8 times higher than that in women (P < 0.0001). Salivary testosterone levels in both sexes negatively correlated with age. The present assay would also be useful in measuring salivary testosterone levels in clinical laboratories.

Analysis of erectile dysfunction drugs and their analogues in counterfeit drugs and herbal medicines by LC-ESI-MS/MS

  • Lee, Ji Hyun;Jeong, Ji Hye;Park, Hyoung-Joon;Do, Jung Ah;Heo, Seok;Cho, Sooyeul;Yoon, Chang-Yong
    • Analytical Science and Technology
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    • v.29 no.4
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    • pp.155-161
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    • 2016
  • Distribution of various illegal or counterfeit drugs of seven approved erectile dysfunction drugs and their analogues has been increased, causing health problems such as cardiovascular disorder, tachycardia, headache, or vision disturbance. We used liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) to determine the erectile dysfunction drugs and their analogues in various counterfeit drugs. Eleven erectile dysfunction drugs and their analogues were detected, with sildenafil and its analogues being the most counterfeited compounds (73.8 %), followed by tadalafil and its analogues (25.4 %). The limits of detection (LOD) and the limits of quantitation (LOQ) of liquid-type and solid-type negative samples ranged from 0.1 to 3.3 ng/mL or ng/g and from 0.3 to 10.0 ng/mL or ng/g, respectively. The recoveries ranged from 84.3 to 112.3 % and 83.2 to 110.2 %, respectively. The contents of sildenafil and tadalafil in the various counterfeit drugs ranged from 21.0 to 947.5 mg/g and from 0.2 to 170.2 mg/g, respectively.

A Successful Regeneration from Shoot Tips of Chrysanthemum morifolium (Ramat.) following Cryopreservation by Droplet-vitrification

  • Yi, Jung-Yoon;Balaraju, Kotnala;Baek, Hyung-Jin;Yoon, Mun-Seop;Kim, Haeng-Hoon;Lee, Young-Yi
    • Korean Journal of Plant Resources
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    • v.31 no.6
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    • pp.675-683
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    • 2018
  • This study describes an efficient and widely applicable droplet-vitrification following cryopreservation for shoot tips of Chrysanthemum morifolium (Ramat.) cvs. 'Borami' and 'Yes morning'. The shoot tips of Chrysanthemum were precultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.7 M). Precultured explants were treated with loading solution (LS, C6) containing glycerol 20% and sucrose 20% for 30 min and exposed to dehydration solution (B5) containing 40% of glycerol and 40% of sucrose for 60 min at $25^{\circ}C$, and then transferred onto droplets containing $2.5{\mu}l$ PVS3 on sterilized aluminum foils ($4cm{\times}0.5cm$) prior to direct immersion in liquid nitrogen (LN) for 1 h. The highest regeneration rate (%) was obtained when shoot tips were precultured with treatment-2 (exposing of shoot tips to MS + 0.3M sucrose for 30 h and then treated with MS+0.5 M sucrose for 16 h) at $25^{\circ}C$ in both the cultivars. The viability of cooled samples, followed by culturing on $NH_4NO_3$-free MS medium for first 5 days was increased to two-fold (80.7%) regrowth rate over those cultured on normal MS medium or MS medium containing plant growth regulators. This result shows droplet-vitrification would be a promising method for cryobanking chrysanthemum germplasm.

Simultaneous Quantitative Determination of Nine Hallucinogenic NBOMe Derivatives in Human Plasma Using Liquid Chromatography Tandem Mass Spectrometry

  • Seo, Hyewon;Yoo, Hye Hyun;Kim, Young-Hoon;Hong, Jin;Sheen, Yhun Yhong
    • Mass Spectrometry Letters
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    • v.10 no.1
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    • pp.18-26
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    • 2019
  • We developed a bioanalytical method for simultaneous determination of nine NBOMe derivatives (25H-NBOMe, 25B-NBOMe, 25E-NBOMe, 25N-NBOMe, 25C-NBOH, 25I-NBOH, 25B-NBF, 25C-NBF, and 25I-NBF) in human plasma using liquid chromatography tandem mass spectrometry (LC-MS/MS). Human plasma samples were pre-treated using solid-phase extraction. Separation was achieved on a C18 column under gradient elution using a mobile phase containing 0.1% formic acid in acetonitrile and 0.1% formic acid in water at a flow rate of 0.3 mL/min. Mass detection was performed in the positive ion mode using multiple reaction monitoring. The calibration range was 1-100 ng/mL for all quantitative analytes, with a correlation coefficient greater than 0.99. The intra- and inter-day precision and accuracy varied from 0.85 to 6.92% and from 90.19 to 108.69%, respectively. The recovery ranged from 86.36 to 118.52%, and the matrix effects ranged from 27.09 to 99.72%. The stability was acceptable in various conditions. The LC-MS/MS method was validated for linearity, accuracy, precision, matrix effects, recovery and stability in accordance with the FDA guidance. The proposed method is suitable for reliable and robust routine screening and analysis of nine NBOMe derivatives in forensic field.