• Title/Summary/Keyword: Protein synthesis

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Molecular Cloning and Characterization of Salt-inducible Aldolase from Salicornia herbacea (퉁퉁마디로부터 염에 의하여 유도되는 Aldolase 유전자의 분리 및 발현분석)

  • Cha, Joon-Yung;Netty Ermawati;Kim, Soon-Gil;Lee, Jeung-Joo;Lim, Chae-Oh;Chung, Woo-Sik;Lee, Kon-Ho;Son, Dae-Young
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.323-328
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    • 2003
  • Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marshes and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display. Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86% identity with Arabidopsis and 78% with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCl concentrations.

Characterization of ent-Kaurenoic Acid 13-Hydroxylase in Steviol Biosynthesis of Stevia rebaudiana Bertoni (Stevia rebaudiana Bertoni의 Steviol 생합성 효소 ent-Kaurenoic Acid 13-Hydroxylase의 특성)

  • Shibata, Hitoshi;Kim, Keun-Ki
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.501-507
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    • 1997
  • Chloroplasts isolated from Stevia rebaudiana Bertoni leaves contained an enzyme activity which catalyzed hydroxylation of ent-kaurenoic acid (ent-kaur-16-en-19-oic acid; ent-KA) to steviol (ent-13-hydroxy kaur-16-en-19-oic acid), the diterpenoid carboxylic alcohol which is the aglycone of sweet stevioside-related glycosides. $[^(14)C]-methylated$ ent-KA was used to localize ent-KA hydroxylase. $[^(14)C]-methyl-KA$ was most actively was transformed into methyl-steviol in chloroplast. The enzymatic activity was found in stroma fraction but not in thylakoid membrane in Stevia rebaudiana Bertoni. However, ent-KA 13-hydroxylase activity was not detected in stroma fraction of either Spinacia oleracea and Solidago altissima. The reaction products using $[^(14)C]-methyl-KA$ were purified and identified on TLC autoradiogram. The hydroxylation of ent-KA from stromal protein to form steviol required NADPH and oxygen. FAD and riboflavin stimulated the enzyme activity 1.5-and 1.7-fold, respectively. It also turned out that the activity of this enzyme using methyl-KA as a substrate was 16.7% that of ent-KA. The purified ent-KA 13-hydroxylase did not act on t-cinnamic acid, 4-hydroxyphenyl acetic acid, choline and resorcinol, known as monooxygenase and hydroxylase substrates.

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Uptake, Assimilation and Translocation of Ammonium or Nitrate in Italian Ryegrass

  • Kim, Tae-Hwan;Lee, Bok-Rye;Jung, Woo-Jin;Kim, Dae-Hyun;Chung, Soon-Ju;Kim, Kil-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.4
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    • pp.303-308
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    • 2001
  • To investigate the partitioning of newly absorbed N derived from NO$_3$- and NH$_4$$^{+}$, 6 mM $K^{15}$ NO$_3$ or 3 mM ($^{15}$ NH$_4$)$_2$ was fed continuously in Italian ryegrass (Lolium multiflrum L.) for 7 days. Nitrogen metabolites (nitrate, amino acid, soluble- and insoluble protein) were analyzed at the end of $^{15}$ N feeding. Dry weight in shoot, stubble and root was not significantly different between NO$_3$$^{[-10]}$ and NH$_4$$^{+}$ feeding. Total nitrogen content in all three organs was significantly higher in NH$_4$$^{+}$ than NO$_3$$^{[-10]}$ feeding. Sum on N content in reduced N fractions (amino acids + proteins) in shoot, stubble and roots in NH$_4$$^{+}$ feeding increased by 13.3, 12.5 and 35.4 %, respectively, compared to NO$_3$$^{[-10]}$ feeding. The Relative Specific Activity (RSA, percentage of newly absorbed $^{15}$ N relative to total N in a sample) values of amino acids and insoluble proteins were significantly higher in NH$_4$$^{+}$ feeding. Total amount of newly absorbed $^{15}$ N in NO$_3$$^{[-10]}$ and NO$_3$$^{[-10]}$ feeding was 52.3 and 69.5 mg/plant on dry matter basis, respectively. In both NH$_4$$^{+}$ and NO$_3$$^{[-10]}$ grown plants, most of the N was allocated to the shoot, 67.5% in NH$_4$$^{+}$ feeding and 58.8% NO$_3$$^{[-10]}$ feeding, respectively. The $^{15}$ N amount incorporated in the reduced N compounds (amino acids and proteins) in NH$_4$$^{+}$ grown plants significantly increased by 74.8% compared to NO$_3$$^{[-10]}$ grown plants. The increase of the $^{15}$ N amount assimilated to amino acids in NH$_4$$^{+}$ grown plants was remarkably higher in roots as more than 7.25 times compared to NO$_3$$^{[-10]}$ feeding. These results indicated that Italian ryegrass was much efficiently utilized NH$_4$$^{+}$-N for the synthesis of reduced N compounds.reduced N compounds.

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THE CYTOTOXIC EFFECTS OF GLASS-IONOMER CEMENT LINERS ON FIBROBLASTS IN HUMAN PULP (Glass-ionomer Cement 이장재의 세포독성에 관한 연구)

  • Na, Young-Min;Min, Byung-Soon;Choi, Ho-Young;Park, Sang-Jin;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.18 no.2
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    • pp.261-276
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    • 1993
  • The purpose of this study was to evaluate for the cytotoxicity of glass-ionomer cement liners(GC liningcement, Ketac-bond, Vitrebond and Fuji lining LC) on the fibroblasts cultured from human pulp. The fibroblasts were cultured in DMEM-10% FBS medium. The measurement of pH, succinate dehydrogenase (SDH) activity test and $^{51}Chromium$ release test were performed. Viable cell count and $^{14}C$-leucine incorporation rate were evaluated following culture time of 2, 4 and 6 days. The results of this study were as follows : 1. The pH in all cements was to be neutralized as time elapsed, and Fuji lining LC was the lowest pH value among them. 2. SDH activity was more inhibited in GC lining cement and Vitrebond than Ketac-bond and Fuji lining LC with the setting process, and GC lining cement and Ketac-bond were reduced after 5 minute's setting and then elevated as time elapsed. 3. In SDH activity test following exposure time, the activity in Vitrebond, GC lining cement and Fuji lining LC was inhibited with increased exposure time, but it was fairly constant in Ketac-bond. 4. Overall the liquid component was more inhibited than the powder component of glass-ionomer cement in SDH activity test. 5. In $^{51}Cr$-release test, Fuji lining LC was the most released of all the cements tested and followed by : Vitrebond, Ketac-bond, GC lining cement. 6. In viable cell count, the number of cells increased as the culture day proceeded in Ketac-bond, but they decreased in GC lining cement. Fuji lining LC was only observed after 2 days culture and there was not observed the whole culture days in Vitrebond. 7. In $^{14}C$-leucine incorporation rate test, protein synthesis was decreased with the number of culture days in GC lining cement, Vitrebond and Fuji lining LC, but it was followed that of control in Ketacbond.

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Purification and Characterization of Exo-xylanase from Escherichia coli Cells Harboring the Recombinant Plasmid pMGl (재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus Exo-xylanase의 정제 및 특성)

  • 문애란;최용진
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.574-582
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    • 1992
  • Exo-xylanase encoded by the xylA gene of Bacillus stearothermoPhillus was produced from Escherichia coli ]M109 carrying a recombinant plasmid pMGL Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at $37^{\circ}C$ for 8 hours on the medium containing 0.5% fructose, 1.0% tryptone, 1.0% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-IOO gel filtration, and Sephadex G-150 gel filtration. The' purified enzyme was most active at pH 6.0 and $45^{\circ}C$. $Ca^{2+}$ and $Co^{2+}$ activated the exo-xylanase activity by about 20% while $Ag^{2+}$, $Fe^{2+}$, $Mg^{2+}$ and $Zn^{2+}$ inhibited the enzyme activity by up to 60%. The $K_m$, value on p-nitrophenyl-$\beta$-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 daL SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 da!. polypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.

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A Significance of Estimation of Proliferating Cell Nuclear Antigen in Thyroid Nodule (갑상선 결절에서 PCNA 측정의 의의)

  • Kim Jung-Chul;Yoon Jung-Han;JeGal Young-Jong
    • Korean Journal of Head & Neck Oncology
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    • v.10 no.2
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    • pp.200-205
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    • 1994
  • Proliferating cell nuclear antgen(PCNA) plays an important role in DNA synthesis in nucleoli and is highly conserved non-histone nuclear protein composed of 261 amino acid. and is considered to correlated with the cells proliferative state, because it is synthesized particulary during the proliferative period of late Gland S-phase. Therefore, PCNA index meaningfully increases in the active or proliferative kinetic cells. By the use of recently developed monoclonal antibodies against PCNA, the immunohistochemical staining methods can make possible. These staining methods are the useful and productive one for ascertaining the cell's proliferating abillity. Moreover, immunohistochemical staining method with a antiPCNA antibody has particulrar advantages as follows. By means of these methods, we can stain the tissue that was already fixed in formalin or paraffin wax. We can see with naked eye that which cell is, where is differentiated through a microscope. Lastly, it maintains the whole tissue architecture and makes a search for the correlation. As we have seen above, the immunohistochemical staining methods for PCNA have been studied as an impotant factor that can find the cell proliferative kinetics in malignancy and biologic behavior of tumors. To investigate of the proliferative activity in thyroid nodule, Authors evaluated cell proliferative activity by immunostaing for PNCA in 45 pathologically confirmed solitary thyroid nodule. The results were as follows. 1) The benign nodules were 25 cases(Adenomatous Goiter: 20 cases, Follicular adenoma: 5 cases) and malignant nodules were 20 cases(Papillary Ca : 14 cases, Follicular Ca : 4 cases, Anaplastic Ca : 2 cases). 2) The Most prevalent age groups were 4th decade(11 cases), and the next group was 5th decade. 3) The average PCNA labelling indices were as follows. Adenomatous goiter(I6.9%), Follicular adenoma(37.6%), papillary Ca(26.3%), Follicular Ca(8.8%) and Anaplastic Ca(86.7%). There were no significant differences in benign(20.4) and malignant nodules (28.8%) except anaplastic Ca(p=0.3226). 4) When the average tumor size 2cm in papillary Ca, the PCNA indices were 26.0% (below 2cm) : 26.6% (above 2cm) (p=0.9642). The PCNA incidies were 23.9% (with lymphatic spread) : 28.7% (without lymphatic spread) (p=0.7056). There were no signlficant differences in the above cases. In conclusion, there were no significant differences in cell proliferative activity by staining for PCNA between benign and malignat nodules except anaplastic Ca.

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Anti-inflammatory effects of a mixture of coffee and sword bean extracts (커피와 작두콩 추출물의 혼합에 따른 항염증 효과)

  • Bae, Hun Cheon;Park, Jung Up;Moon, Jae-Hak
    • Korean Journal of Food Science and Technology
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    • v.52 no.3
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    • pp.237-243
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    • 2020
  • Coffee is one of the most widely consumed beverages in the world, and sword bean (Canavalia gladiata, SB) reportedly possesses various biological activities. Therefore, in this study, to reduce caffeine intake and improve coffee function, SB was selected as a supplementary material for blending coffee. The antioxidant and anti-inflammatory activities of coffee with the SB extract at concentrations of 0.1-0.5% (v/v) were evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and RAW 264.7 cells, respectively. The DPPH radical-scavenging activity of SB-treated coffee depended on the concentration of the SB extract. In the cell culture experiment, cytotoxicity was not observed at any SB concentration. In addition, the inducible nitric oxide synthesis protein expression as well as the increases in nitric oxide and interleukin-6 expression were effectively inhibited by SB addition to the coffee. These results indicate that SB might be useful as a supplementary ingredient to enhance the caffeinated drink functions.

Effect of Diallyl Disulfide on Heme Oxygenase-1 Expression in Human Hepatoma Cell Line HepG2 (인간 간암세포주 HepG2에서 heme oxygenase-1 발현에 대한 diallyl disulfide의 효과)

  • Kim, Kang-Mi;Lee, Sang-Kwon;Park, Young-Chul
    • Journal of Life Science
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    • v.21 no.7
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    • pp.1046-1051
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    • 2011
  • Diallyl disulfide (DADS), the most prevalent oil-soluble organosulfur compound in garlic, is known to have diverse biological activities, including anticarcinogenic, antiatherosclerotic, antiinflammatory, and antioxidant actions. In this study, we investigated the effect of DADS on the expression of heme oxygenase-1 (HO-1) in human liver hepatoma cell line HepG2. Treatment of HepG2 cells by DADS evoked a dose-dependent growth inhibition without significant toxicity to the cells, and also induced the expression of transcription factor Nrf2. However, DADS did not have any enhancing effect on transcription and translation of HO-1 expression in HepG2 cells. In addition, DADS efficiently blocked protein synthesis of HO-1 in HepG2 cells stimulated by CoPP or hemin. But, DADS did not decrease the content of transcripts of HO-1 gene stimulated by CoPP, with accumulation of Nrf2 and small Maf in the nucleus. Based on these results, we conclude that DADS inhibits HO-1 expression by modulation of translational level of CoPP or hemin-induced HO-1 expression in HepG2 cells.

The Metabolic Effects of FGF21: From Physiology to Pharmacology (생리, 약학적 관점에서 fibroblast growth factor 21 (FGF21)의 대사 효과 고찰)

  • Song, Parkyong
    • Journal of Life Science
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    • v.30 no.7
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    • pp.640-650
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    • 2020
  • Fibroblast growth factor 21 (FGF21) is an atypical member of the FGF protein family which is highly synthesized in the liver, pancreas, and adipose tissue. Depending on the expression tissue, FGF21 uses endo- or paracrine features to regulate several metabolic pathways including glucose metabolism and energy homeostasis. Different physiologically stressful conditions such as starvation, a ketogenic diet, extreme cold, and mitochondrial dysfunction are known to induce FGF21 synthesis in various tissues to exert either adaptive or defensive mechanisms. More specifically, peroxisome proliferator-activated receptor gamma and peroxisome proliferator-activated receptor alpha control FGF21 expression in adipose tissue and liver, respectively. In addition, the pharmacologic administration of FGF21 has been reported to decrease the body weight and improve the insulin sensitivity and lipoprotein profiles of obese mice and type 2 diabetes patients meaning that FGF21 has attracted huge interest as a therapeutic agent for type 2 diabetes, obesity, and non-alcoholic fatty liver disease. However, understanding FGF21 remains complicated due to the paradoxical condition of its tissue-dependent expression. For example, nutrient deprivation largely increases hepatic FGF21 levels whereas adipose tissue-derived FGF21 is increased under feeding condition. This review discusses the issues of interest that have arisen from existing publications, including the tissue-specific function of FGF21 and its action mechanism. We also summarize the current stage of a clinical trial using several FGF21 analogs.

Effects of Aspirin on the Pathogenesis of Acute Lung Injury in Rats Subjected to Hemorrhage (출혈성 쇼크로 인한 급성 폐손상의 발병기전과 아스피린의 효과)

  • Park, Yoon-yub;Lee, Young Man
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.1
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    • pp.83-91
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    • 2006
  • Background : For unknown reasons, the serum ferritin concentrations are higher in patients with acute lung injury. A pretreatment with aspirin reduces the acute lung injury in rats subjected severe hemorrhage, and increases the rate of ferritin synthesis in vitro. This study investigated the effect of aspirin on the serum ferritin changes in rats subjected to severe hemorrhage. Methods : Hemorrhagic shock was induced by withdrawing blood (20 ml/kg of B.W.) through the femoral artery for 5 min. The rats were pretreated with aspirin (10 mg/kg, i.v.) 30 min before hemorrhage. Results : The protein content and leukocyte count in the bronchoalveolar lavage fluid, lung tissue myeloperoxidase activities were significantly higher after hemorrhage. The aspirin pretreatment prevented these changes. The serum and lavage fluid ferritin concentrations were elevated higher after hemorrhage. These were also attenuated by the aspirin pretreatment. Conclusion : The changes in the serum and lung lavage ferritin level might be closely related to the severity of hemorrhage-induced acute lung injury. Therefore, the serum and lavage ferritin concentrations can be a useful biomarker for patients with precipitating conditions.