• Title/Summary/Keyword: MG/MS

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Influences by position of node and existence of leaf on microtuberization in node culture of potato (감자절간배양에서 마디위치 및 잎의 부착유무에 따른 소괴경형성의 차이)

  • Hwang, Hye-Yeon;Lee, Young-Bok
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.63-68
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    • 2008
  • Single-node stem pieces ca. 1 cm in length containing a axillary bud were obtained from in vitro plants of potato (Solanum tuberosum L.). The influences by a position of the node and the existence of a leaf at the node were observed in the single-node culture on the 8% sucrose MS medium. The effect of CCC was also investigated for the microtuberization. The apical part node was excellent in the tuberization not to mention shoot length, fresh weight, diameter, the number of node on the in vitro culture of a single-node than the lower part. The differences in the diameter of a tuber formed in the part of the axillary bud on all treatments including the cultivation of the apical part node were not recognized. However, the fresh weight of the tuber showed high value in the tuber formed at the axillary bud of shoot apex part. At 20 days after cultivation, tuberization was promoted in the new stolen that developed from the bud of node with a leaf under SD condition of 8 hours at $20^{\circ}C$. The tuberization from axillary bud of the single-node without leaf was inhibited at high temperature of $28^{\circ}C$ regardless of daylength. Whereas, tuberization at $20^{\circ}C$ and $28^{\circ}C$ was similar without the difference under SD condition but the tuber formation ratio were low. CCC 500 mg/L promoted tuberization and the effect was also showed even under LD condition at $28^{\circ}C$. The inhibiton of tuberization under LD and high temperature condition could be solved by treatment with CCC.

Aluminum toxicity-induced alterations of root proteome in wheat seedlings

  • Oh, Myeong Won;Roy, Swapan Kumar;Cho, Kun;Cho, Seong-Woo;Park, Chul-Soo;Chung, Keun-Yook;Choi, Jong-Soon;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.127-127
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    • 2017
  • Aluminum is the most abundant metallic element in the Earth's crust and considered as the most limiting factor for plant productivity in acidic soils. The inhibition of root growth is recognized as the primary effect of Al toxicity. Seeds of wheat cv. Keumkang (Korean cultivar) were germinated on petridish for 5 days and then transferred hydroponic apparatus which was treated with $0{\mu}M$ $AlCl_3$ (control), $100{\mu}M$ $AlCl_3$ and $150{\mu}M$ $AlCl_3$ for 5 days. The length of roots, shoots and fresh weight of wheat seedlings were decreased under aluminum stress. The concentrations of $K^+$, $Mg^{2+}$ and $Ac^{2+}$ were decreased whereas $Al^{3+}$ and $P_2O_5{^-}$ concentration was increased under aluminum stress. Using confocal microscopy, the fluorescence intensity of aluminum was increased with morin staining. In this study, a proteome analysis was performed to identify proteins, which is responsible to aluminum stress in wheat roots. In 10-day-old seedlings, proteins were extracted from roots and separated by 2-DE, stained by CBB. Using image analysis, a total of 47 differentially expressed protein spots were selected, whereas 19 protein spots were significantly up-regulated such as s-adenosylmethionine, oxalate oxidase, malate dehydrogenase, cysteine synthase, ascorbate peroxidase and 28 protein spots were significantly down-regulated such as heat shock protein 70, o-methytransferase 4, enolase, amylogenin by aluminum stress following protein spots analyzed by LTQ-FTICR mass spectrometry. The results provide the global picture of Al toxicity-induced alterations of protein profiles in wheat roots, and identify the Al toxicity-responsive proteins related to various biological processes that may provide some novel clues about plant Al tolerance.

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Development of Automatic Nutrient-Solution Mixing System Using a Low-Cost and Precise Liquid Metering Device (액제 정밀계량 장치를 이용한 양액 자동조제 시스템 개발)

  • 이규철;류관희;이정훈;김기영;황호준
    • Journal of Biosystems Engineering
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    • v.22 no.4
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    • pp.469-478
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    • 1997
  • This study was conducted to develop an automatic nutrient-solution mixing system for small-scale sewers. The nutrient-solution mixing system consisted of a low-cost and precise metering device and data acquisition & control system with a personal computer. and, the metering device was composed of three parts those were supply pumps, metering cylinders and venturi tube. The system controlled electric conductivity(EC) and pH of nutrient-solution based on the time-based feedback control method with the information about temperature, EC, and pH of the nutrient-solution. The performance of the nutrient-solution mixing system was evaluated through the control of EC and pH while compared with those of commercial system. Also an experimental cultivation of tomato was conducted to verify and to improve the developed system. Results of this study were as follows. 1. The correlation coefficient of meteing device between the flow rate and operating time was 0.9999, and the linear reuession equation computed was y=21.759x, where y is the discharge($g$) and x is the operating time(s). 2. Calculated errors for the developed metering device and two commercial pump were $\pm$0.3% $\pm$2.45% and $\pm$1.38 % FS error respectively. 3. An automatic nutrient-solution mixing system based on a low-cost and precise metering device was developed. 4. The full scale errors of the developed system in controlling EC and pH at 23$\pm$1$^{\circ}C$ were $\pm$0.05mS/cm and $\pm$0.2, respectively 5. When using the commercial system, the controlled values of EC and pH of the 500 $\ell$ of water were 1.29 mS/cm and 6.1 pH for the setting points of 1.4 mS/cm and 6.0 pH respectively at 23$pm1^{\circ}C$. 6. The developed nutrient-solution control system showed $\pm$0.05 ms/cm of deviation from the setting EC value over the experimental cultivation period. 7. The deviation from the average values of Ca and Mg mass content in the several nutrient-solution were 0.5% and 1.8% respectively.

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Effects of replacing fish oil with palm oil in diets of Nile tilapia (Oreochromis niloticus) on muscle biochemical composition, enzyme activities, and mRNA expression of growth-related genes

  • Ayisi, Christian Larbi;Zhao, Jinliang;Yame, Chen;Apraku, Andrews;Debra, Grace
    • Fisheries and Aquatic Sciences
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    • v.22 no.11
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    • pp.25.1-25.9
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    • 2019
  • Background: Due to the continuous demand for fish coupled with decline in capture fisheries, there is the need to increase aquaculture production to meet the demand. Aquaculture is faced with high cost of feeding since fish oil and fish meal are expensive. In view of this, there are calls to explore alternatives that are cheap and reliable. Objectives: This study on Oreochromis niloticus was conducted to evaluate the effects of replacing fish oil (FO) with palm oil (PO) at 0%, 25%, 50%, 75%, and 100% on muscle fatty acid and proximate composition as well as growthrelated enzyme activities and mRNA expression. Methods: Oreochromis niloticus were fed five experimental diets (33% crude protein and 10% crude lipid) for 8 weeks. Feed had variation in fish oil and palm oil contents. After the 8 weeks feeding trial, five fish were sampled from each tank (15 from each treatment) and euthanized using an excess dose of tricaine methane sulfonate (MS-222 at 200 mg/L). Fatty acid and enzyme activities were analyzed using standard protocols. Also, RT-qPCR was used to quantify the expression levels of selected growth-related genes. Results: Fish fed 25% PO recorded the least muscle protein content and was significantly lower than the group fed 100% PO. Paired box protein 7 (Pax-7) enzyme activity was significantly higher in the group fed 50% PO compared to the groups fed 25% PO and 100% PO, while caplain-3 (Capn-3) was significantly lower in the group fed 0% PO compared to all other groups. There was a significant difference among treatments with respect to mRNA expression of Pax-7 and Capn-3. Group fed 25% PO had significantly lower mRNA expression of Pax-7, while the group fed 75% PO recorded significantly higher mRNA expression of Capn-3 compared to groups fed 0% PO, 25% PO, and 100% PO. Pearson's correlation analysis revealed that Igf-I and Igf-II mRNA expression have significant correlation with n-3 polyunsaturated fatty acids content in muscle. Conclusion: The results suggest muscle protein content could be modified if FO is replaced with PO. Also, mRNA expression of Pax-7 and Capn-3 is affected by replacing FO with PO.

The Role of Adenosine Receptor on Acetylcholine Release from Ischemic-Induced Rat Hippocampus (허혈이 유발된 흰쥐 해마에서 Acetylcholine 유리에 미치는 Adenosine 수용체의 역할)

  • Choi, Bong-Kyu;Kim, Do-Kyung;Kang, Hun;Jeon, Jae-Min;Kang, Yeon-Wook
    • The Korean Journal of Pharmacology
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    • v.32 no.2
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    • pp.127-138
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    • 1996
  • The effects of adenosine analogues on the electrically-evoked acetylcholine(ACh) release and the influence of ischemia on the effects were studied in the rat hippocampus. Slices from the rat hippocampus were equilibrated with $0.1{\mu}M$ $[^3H]-choline$ and the release of the labelled product, $[^3H]-ACh$, was evoked by electrical stimulation(3 Hz, 2 ms, 5 $VCm^{-1}$ and rectangular pulses for 2 min), and the influence of various agents on the evoked tritiumoutflow was investigated. Ischemia(10 min with 95% $N_2$ + 5% $CO_2$) increased both the basal and evoked ACh release. These increases were abolished by glucose addition into the superfused medium, and they significantly inhibited either by 0.1 & $0.3{\mu}M$ TTX pretreatment or by removing $Ca^{++}$ in the medium. MK-801($1{\sim}10{\mu}M$), a specific NMDA receptor antagonist, and glibenclamide $(1{\mu}M)$, a $K^+-channel$ inhibitor, did not alter the evoked ACh release and nor did they affect the ischemia-induced increases In ACh release. However, polymyxin B(0.03 mg), a specific protein kinase C inhibitor, significantly inhibited the effects of ischemia on the evoked ACh release. Adenosine and $N^6-cyclopentyladenosine$ decreased the ACh release in a dose dependent manner in ischemic condition, though the magnitude of inhibition was far less than those in normal(normoxic) condition. However, the treatment with $5{\mu}M$ DPCPX, a potent $A_1-adenosine$ receptor antagonist, potentiated the ischemia-effect. These results indicate that the evoked-ACh release is potentiated by ischemia, and this process being most probably mediated by protein kinase C, and that the decreased effect of ACh release mediated by $A_1-adenosine$ receptor is significantly inhibited in ischemic state.

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Anti-obesity effects of cultivated ginseng, -wild simulated ginseng and -red ginseng extracts (인삼, 산양산삼 및 홍삼 추출물의 항비만 효과)

  • Kim, Ha-Rim;Lee, Chang-Hyun;Jung, Mun Yhung;Kim, Jong-Seok;Kim, Hong-Jun;Jeon, Hye-Sook;Lee, Seung-Hyun;Kim, Ji-Hyun;Shin, Min-Ji;Ma, Sang-Yong;Kwon, Jin;Oh, Chan-Ho
    • Herbal Formula Science
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    • v.27 no.4
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    • pp.269-284
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    • 2019
  • This study was to investigate whether cultivated ginseng (CG), cultivated wild simulated ginseng (CWG) and cultivated red ginseng (CRG) extracts influences on the obesity. The saponin contents of 3 kinds of ginsengs were analysed by HPLC-ESI-TOF-MS. Total saponin contents were determined in CG on the most contents and since red ginseng has the highest PD (protopanaxadiol type) / PT (protopanaxatriol type) ratio, there may be differences between ginseng, wild ginseng, and red ginseng with respect to their pharmacological effects. Male C57BL/6J mice were fed a normal diet(N), HFD (60% Kcal fat, C), HFD with CG, CWG and CRG extracts (800 mg/kg) for 5 weeks. We observed change of total body weight, degree of hepatic lipid accumulation and immunohistochemical change of GLP-1 and insulin-secreting cells. Also this study attempts to use the physiological analysis method to analyze the changes of blood lipids, insulin and leptin concentration. The change of body weight and size of accumulated lipid droplets in liver lobules decreased in all of the experimental groups than the control(C) group. In the pancreas, the immunohistochemical density of insulin-secreting cells were significantly stronger in the CWG and CRG than C group. The levels of serum insulin and leptin significantly decreased 55.6%, 54.3% respectively in CWG and CRG. The changes of triglyceride, total cholesterol in serum decreased in CRG than the C group. Obesity related CG, CWG and CRG extracts might have contribute to improvement of obesity by regulating the levels of blood lipids and biochemical indicator of fat accumulation.

In vitro Variant Induction and Its Content of Gentiopicroside of Gentiana scabra BUNGE (용담(龍膽)의 기내변이주(器內變異株) 유도(誘導)와 변이주(變異株)의 Gentiopicroside 함량(含量))

  • Seong, Nak-Sul;Park, Chung-Heon;Kim, Kwan-Su;Lee, Seoung-Tack;Chang, Yeong- Hee
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.1
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    • pp.40-44
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    • 1995
  • This study was aimed to mutant induction in Gentiana scabra BUNGE through the tissue culture techniques and comparison of gentiopicroside content in each mutant plants derived from tissue culture. In order to induce the mutants of Gentiam scabra, seed, apical and lateral bud were soaked in $NaN_3$, EMS and MNU solutions and cultured on MS basal medium containing 0.5mg/l NAA, BA $NaN_3$, MNU-treated buds survived about 50% but not survived in EMS treatment. Seed germination rate was extremely low in EMS and MNU treatments but various types of mutant plantlets were obtained by those treatments. 63% of elongated type and 37% of dwarf type were obtained from tissue culture treated by various mutagen. Gentiopicroside contents of regenerated plantlets was analyzed. Root of dwarf type contained more gentiopicroside(1.383%) than that of elongation type (0.083%).

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Purification and Characterization of Gibberellin $3Beta$-Hydroxylase from Immature Seeds of Phaseolus vulgaris (강낭콩미숙종자로부터 Gibberellin $3Beta$-Hydroxylase 정제 및 성질)

  • 곽상수
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.133-148
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    • 1987
  • Gibberellin(GA) 3-$\beta$ hydroxylation is very important for the shoot elogation in the higher plants, since only 3$\beta$-hydryoxylated GAs promote shoot elogation in several plants. Fluctuation of 3$\beta$-hydryoxylase activity was examined during seed maturation using two cultivars of , P. vulgaris, Kentucky Wonder (normal) and Masterpiece (dwarf). Very immature seeds of both cultivars contain high level of 3$\beta$-hydroxylase activity (per mg protein). Both cultivars showed maximum of enzyme activity (per seed) in the middle of their maturation process. Gibberellin 3$\beta$-hydroxylase catalyzing the hydroxylation of GA20 to GA1 was purified 313-fold from very early immature seeds of P. vulgaris. Crude soluble enzyme extracts were purified by 15% methanol precipitation, hydrophobic interaction chromatogrphy, DEAE ion exchange column chromatography and gel filtration HPLC. The 3$\beta$-hydroxylase activity was unstable and lost much of its activity duting the purification. The molecular weight of purified enzyme was extimated to be 42, 000 by gel filtration HPLC and SDS-PAGE. The enzyme exhibited maximum activity at pH 7.7. The Km values for [2.3-3H] GA20 and [2.3-3H]GA9 were 0.29 $\mu$M and 0.33 $\mu$M, respectively. The enzyme requires 2-oxoglutarate as a cosubstrate; the Km value for 2-oxoglutarate was 250 $\mu$M using 3H GA20 as a substrate. Fe2+ and ascorbate significantly activated the enzyme at all purification steps, while catalase and BSA activated the purified enzyme only. The enzyme was inhibited by divalent cations Mn2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+ and Hg2+. Effects of several GAs and GA anaogues on the putrified 3$\beta$-hydroxylase were examined using [3H]GA9 and GA20 as a substrates. Among them, GA5, GA9, GA15, GA20 and GA44 inhibited the enzyme activity. [13C, 3H] GA20 was converted by the partially purified enzyme preparation to [13C, 3H]GA1, GA5 and GA6, which were identified by GC-MS, GA9 was converted only GA4, GA15 and GA44 were converted to GA37 and GA38, respectively. GA5 was epoxidized to GA6 by the preparation. This suggests that 3$\beta$-hydroxylation of GA20 and epoxidation of GA5 are catalyzed by the same enzyme in P, vulgaris.

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Cloning, Over-expression, and Characterization of YjgA, a Novel ppGpp-binding Protein

  • Gnanasekaran, Gopalsamy;Pan, SangO;Jung, Wontae;Jeong, Kwangjoon;Jeong, Jae-Ho;Rhee, Joon Haeng;Choy, Hyon E.;Jung, Che-Hun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.8
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    • pp.2419-2424
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    • 2013
  • Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

Studies on the Analysis of Benzo(a)pyrene and Its Metabolites on Biological Samples by Using High Performance Liquid Chromatography/Fluorescence Detection and Gas Chromatography/Mass Spectrometry

  • Lee, Won;Shin, Hye-Seung;Hong, Jee-Eun;Pyo, Hee-Soo;Kim, Yun-Je
    • Bulletin of the Korean Chemical Society
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    • v.24 no.5
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    • pp.559-565
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    • 2003
  • An analytical method the determination of benzo(a)pyrene (BaP) and its hydroxylated metabolites, 1-hydroxybenzo(a)pyrene (1-OHBaP), 3-hydroxybenzo(a)pyrene (3-OHBaP), benzo(a)pyrene-4,5-dihydrodiol (4,5-diolBaP) and benzo(a)pyrene-7,8-dihydrodiol (7,8-diolBaP), in rat urine and plasma has been developed by HPLC/FLD and GC/MS. The derivatization with alkyl iodide was employed to improve the resolution and the detection of two mono hydroxylated metabolites, 1-OHBaP and 3-OHBaP, in LC and GC. BaP and its four metabolites in spiked urine were successfully separated by gradient elution on reverse phase ODS $C_{18}$ column (4.6 mm I.D., 100 mm length, particle size 5 ㎛) using a binary mixture of MeOH/H₂O (85/15, v/v) as mobile phase after ethylation at 90 ℃ for 10 min. The extraction recoveries of BaP and its metabolites in spiked samples with liquid-liquid extraction, which was better than solid phase extraction, were in the range of 90.3- 101.6% in n-hexane for urine and 95.7-106.3% in acetone for plasma, respectively. The calibration curves has shown good linearity with the correlation coefficients (R²) varying from 0.992 to 1.000 for urine and from 0.996 to 1.000 for plasma, respectively. The detection limits of all analytes were obtained in the range of 0.01-0.1 ng/mL for urine and 0.1-0.4 ng/mL for plasma, respectively. The metabolites of BaP were excreted as mono hydroxy and dihydrodiol forms after intraperitoneal injection of 20 mg/kg of BaP to rats. The total amounts of BaP and four metabolites excreted in dosed rat urine were 3.79 ng over the 0-96 hr period from adminstration and the excretional recovery was less than 0.065% of the injection amounts of BaP. The proposed method was successfully applied to the determination of BaP and its hydroxylated metabolites in rat urine and plasma for the pharmacokinetic studies.