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Preparation of minor ginsenosides C-Mc, C-Y, F2, and C-K from American ginseng PPD-ginsenoside using special ginsenosidase type-I from Aspergillus niger g.848

  • Liu, Chun-Ying;Zhou, Rui-Xin;Sun, Chang-Kai;Jin, Ying-Hua;Yu, Hong-Shan;Zhang, Tian-Yang;Xu, Long-Quan;Jin, Feng-Xie
    • Journal of Ginseng Research
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    • v.39 no.3
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    • pp.221-229
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    • 2015
  • Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.

Clues to Understand the Regulatory Metabolisms Governing the Biosynthesis of Fungal Secondary Metabolites

  • Choi, Yoon-E
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.14-14
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    • 2015
  • Fungi are of particular interest due to their capacity to produce an extensive array of secondary metabolites. While many secondary metabolites have no known functions to the producing fungal organisms, these metabolites have tremendous importance to humans with beneficial (e.g., antibiotics) or detrimental (e.g., mycotoxins) properties. In this study, two important filamentous fungi, Fusarium verticillioides and Mycosphaerella graminicola were selected as target species and the genes regulatory functions on the biosynthesis of secondary metabolisms were studied. Functional genomics including forward and reverse genetics, and proteomics were utilized to better understand the complex secondary metabolism regulations in both F. verticillioides and M. graminicola. Identified genes in either F. verticillioides or M. graminicola background were CPP1 (a putative protein phosphatase gene), GAC1 (encoding a GTPase activating protein), MCC1(encoding c-type cyclin), and the velvet gene, MVE1. Our data suggest that there are diverse regulatory genes on fungal secondary metabolites with distinct or overlapping functional roles.

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Genetic Polymorphism of Vascular Endothelial Growth Factor (VEGF C936T) in the Korean Population

  • Kim, Jin-Kyeoung;Oh, Do-Yeun;Kwak, Sun-Young;Han, Jin-Hee;Chung, Young-Sun;Kim, Nam-Keun
    • Animal cells and systems
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    • v.7 no.3
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    • pp.261-264
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    • 2003
  • Leiden), no mutation was detected in either group. Allele frequencies of A2379G and G2391 A mutations were not significantly different between CAD patients and controls. Non-Caucasian populations have a considerably lower factor Ⅴ Leiden allele frequency than Caucasian populations. Thus, it may be due to differences in the genetic background as well as environmental factors.

Vasodilator-stimulated phosphoprotein-phosphorylation by ginsenoside Ro inhibits fibrinogen binding to αIIb/β3 in thrombin-induced human platelets

  • Shin, Jung-Hae;Kwon, Hyuk-Woo;Cho, Hyun-Jeong;Rhee, Man Hee;Park, Hwa-Jin
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.359-365
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    • 2016
  • Background: Glycoprotein IIb/IIIa (${\alpha}aIIb/{\beta}_3$) is involved in platelet adhesion, and triggers a series of intracellular signaling cascades, leading to platelet shape change, granule secretion, and clot retraction. In this study, we evaluated the effect of ginsenoside Ro (G-Ro) on the binding of fibrinogen to ${\alpha}aIIb/{\beta}_3$. Methods: We investigated the effect of G-Ro on regulation of signaling molecules affecting the binding of fibrinogen to ${\alpha}aIIb/{\beta}_3$, and its final reaction, clot retraction. Results: We found that G-Ro dose-dependently inhibited thrombin-induced platelet aggregation and attenuated the binding of fibrinogen to ${\alpha}aIIb/{\beta}_3$ by phosphorylating cyclic adenosine monophosphate (cAMP)-dependently vasodilator-stimulated phosphoprotein (VASP; $Ser^{157}$). In addition, G-Ro strongly abrogated the clot retraction reflecting the intensification of thrombus. Conclusion: We demonstrate that G-Ro is a beneficial novel compound inhibiting ${\alpha}aIIb/{\beta}_3$-mediated fibrinogen binding, and may prevent platelet aggregation-mediated thrombotic disease.

VAD By Neural Network Under Wireless Communication Systems (Neural Network을 이용한 무선 통신시스템에서의 VAD)

  • Lee Hosun;Kim Sukyung;Park Sung-Kwon
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.30 no.12C
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    • pp.1262-1267
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    • 2005
  • Elliptical basis function (EBF) neural network works stably under high-level background noise environment and makes the nonlinear processing possible. It can be adapted real time VAD with simple design. This paper introduces VAD implementation using EBF and the experimental results show that EBF VAD outperforms G729 Annex B and RBF neural networks. The best error rates achieved by the EBF networks were improved more than $70\%$ in speech and $50\%$ in silence while that achieved by G.729 Annex B and RBF networks respectively.

Nutritional Analysis of Wild Conyza canadensis L. Extract (야생 망초(Conyza canadensis L) 추출물의 영양성분 분석)

  • Kim, Hong Yul;Song, Hyun Sook;Lee, Geo Lyong
    • Journal of Naturopathy
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    • v.11 no.2
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    • pp.100-108
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    • 2022
  • Background: It has been reported that various substances exist in wild Conyza canadensis, but I think that the overall compositional analysis is still insufficient. Purposes: This study was to set extraction conditions and search for various nutritional components in the wild C. canadensis. Methods: Various extracts were extracted and analyzed using different analytical instruments to suggest and study the conditions for component analysis. Results: As for the general nutritional components of C. canadensis leaf extract, carbohydrates were 46.4%, crude protein 26.5%, moisture 16.3%, raw meal 9.5%, and natural fat 1.2%. The dietary fiber content was 30.76%. Free fructose, glucose, sucrose, and maltose were separated, and the amount of fructose was as high as 4,144.2 mg/100 g. In addition, we found 64.87 mg/g of K, 4.09 mg/g of P, 1.77 mg/g of Mg, and 0.16 mg/g of disodium in the extracts. A total of 20 fatty acids have appeared. In addition, unsaturated fatty acids (octadecenoic acid, octadecadienoic acid, and linolenic acid) were detected. Linolenic acid was high at 54.1%. The saturated fatty acid content was 0.39 g/100 g, and the trans fatty acid content was 0.01 g/100 g. No cholesterol was found in the plant. Riboflavin was at 0.4 mg/100 g, with six essential amino acids and 24 amino acids. Conclusions: As a result of the primary research on the plant, various nutritional and functional ingredients exist in the extract, and natural healing uses are available.

Analgesia after Epidural Dexamethasone is Further Enhanced by IV Dipyrone, but Not IV Parecoxibe Following Minor Orthopedic Surgery

  • Lauretti, Gabriela R.;Righeti, Claudia C.F.;Kitayama, Antonio T.
    • The Korean Journal of Pain
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    • v.27 no.4
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    • pp.345-352
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    • 2014
  • Background: Epidural administration of dexamethasone has been suggested for pain control after minor orthopedic surgery. This study was conducted to assess its efficacy after such surgery, combined or not to IV dipyrone, IV parecoxibe or their combination. Methods: 91 patients were randomly assigned to seven groups. Patients were submitted to spinal bupivacaine anesthesia combined to epidural administration of either 10 ml saline or 10 mg dexamethasone diluted to 10-ml volume. Patients also received 10 ml IV saline or 1 gr dipyrone and/or 40 mg parecoxibe diluted to 10 ml with saline. Control group (CG) received epidural and IV saline. Dexamethasone group (DexG) received epidural dexamethasone and IV saline. Dipyrone group (DipG) received epidural saline and IV dipyrone. Dex-Dip G received epidural dexamethasone and IV dipyrone. Parecoxibe group (ParG) received epidural saline and IV parecoxibe. Dex-ParG received epidural dexamethasone and IV parecoxibe. Finally, Dex-Dip-ParG received epidural dexamethasone and IV dipyrone plus IV parecoxibe. Results: The CG expressed 4h of analgesia and sooner requested pain killer. DexG was similar to DipG or ParG or Dex-ParG (7-hours), and they requested less ketoprofen compared to the CG (P < 0.05). However, the Dex-DipG and the Dex-Dip-ParG resulted in longer time to demand pain killer (17-hours) and less ketoprofen consumption in 24-hours (P < 0.002). Adverse effects were similar among groups. Conclusions: The analgesia secondary to epidural dexamethasone was enhanced by IV dipyrone, while no effects were observed by the addition of IV parecoxibe.

Isolation and Structural Identification of Antioxidant Substances from Ethyl Acetate Extract of Conyza canadensis (망초(Conyza canadensis) Ethyl Acetate 추출물의 항산화성 물질의 분리와 동정)

  • Hyun Sook Song
    • Journal of Naturopathy
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    • v.12 no.1
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    • pp.7-15
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    • 2023
  • Background: As a result of analyzing the components of wild Conyza canadensis, it contains physiologically active ingredients, so it is necessary to identify the compound. Purposes: It was to study the compound's molecular structure; a previous study showed that C. canadensis contains antioxidant substances. Methods: The ultrasonic pulverized lysate of C. canadensis stem and leaves was first extracted with 90% methanol and then five organic solvents. Next, the extracts was fractionated by HPLC, LC/MS chromatography, and NMR analyzers identified the molecular structure. Results: 100 g of dry C. canadensis was sonicated in 90% methanol and concentrated under reduced pressure to 11.96 g of a crude extract. Then, this crude was extracted with five types of solvents to obtain 123.8 mg of n-hexane, 448.2 mg of dichloromethane, 1047.7 mg of ethyl acetate (EA), 2563.8 mg of butanol, and 7.04 g of water. The EA extracts were fractionated by LC-MS and then re-fractionated to obtain F1 to F20. Next, the F15 was further fractionated to obtain nine fine fractions. Finally, the F17 fraction was re-fractionated to obtain ten fine fractions. As a result of LC-MS and NMR spectrometer analysis of the F15-7, the structure of this compound was confirmed as 3,5-dicaffeoylquinic acid. As a result of examining the structures of the F17-4 and F17-5 fractions, Quercetin-3-o-β-galactose was identified. In addition, the form of the F17-10 was confirmed to be 1,3,4-tri-caffeoylquinic acid. Conclusions: This study demonstrated that C. canadensis contained phenolic antioxidants, and its utilization may be expected.

Ectopic Expression of Cenexin1 S796A Mutant in $ODF2^{+/-}$ Knockout Background Causes a Sperm Tail Development Defect

  • Lee, Kyung Ho
    • Development and Reproduction
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    • v.16 no.4
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    • pp.363-370
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    • 2012
  • The outer dense fiber 2 (ODF2) protein is an important component of sperm tail outer dense fiber and localizes at the centrosome. It has been reported that the RO072 ES cell derived homozygote knock out of ODF2 results in an embryonic lethal phenotype, and XL169 ES cell derived heterozygote knock out causes severe defects in sperm tail development. The ODF2s splicing variant, Cenexin1, possesses a C-terminal extension, and the phosphorylation of serine 796 residue in an extended C-terminal is responsible for Plk1 binding. Cenexin1 assembles ninein and causes ciliogenesis in early stages of the cell cycle in a Plk1-independent manner. Alternatively, in the late stages of the cell cycle, G2/M phase, Cenexin1 binds to Plk1 and results in proper mitotic progression. In this study, to identify the in vivo function of Plk1 binding to phosphorylated Cenexin1 S796 residue, and to understand the in vivo functional differences between ODF2 and Cenexin1, we generated ODF2/Cenexin1 S796A/Cenexin1 WT expressing transgenic mice in a RO072 ES cell derived $ODF2^{+/-}$ knock out background. We observed a severe defect of sperm tail development by ectopic expression of Cenexin1 S796A mutant and no phenotypic differences between the ectopic expression of ODF2/Cenexin1 WT in $ODF2^{+/-}$ background and in normal wild type mice.