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Effect of Lipoxygenase on the Oxidation of Rainbow Trout Lipid in Model system (모델시스템에 있어서 무지개 송어 지방질의 산화에 대한 Lipoxygenase의 영향)

  • 김혜경;엄수현;최홍식
    • Journal of Life Science
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    • v.5 no.2
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    • pp.70-75
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    • 1995
  • The effect of lipoxygenase (LOX) on the oxidation and co-oxidation of lipid fraction was studied in the model system of rainbow trout. For the reaction in model system 1 g of lipid fraction and 50mL of enzyme extract(LOX, 140 unit in 50mL phosphate buffer solution at pH 7, 4)), which were obtained from rainbow trout, were homoginized in the presence of Tween 20 and kept at 23$\circ$C for 3 days. The activity of LOX was decreased to 43% of initial level during the reaction in the model system. The initial composition of rainbow trout lipid was showed to be consisted of trigliceride(TG;82%) and free fatty acid(FFA;0.1%), while this converted to 59% of TG and 20% of FIFA, respectively after reaction in model system. Change of fatty acid composition was also observed and the content of linoleic acid, one of the major fatte acids, was decreased to 13% from 54% in the content of total fatty acids after reaction. The carotenoids in rainbow trout were composed of 0.4% $\alpha$-carotene, 1.6% $\beta$ -carotene, 80% canthaxanthin, 7% lutein and 11% zeaxanthin, thus the canthaxanthin was the major component. This canthaxanthin was the most degraded carotenoid by lipoxygenase catalyzed co-oxidation during the reaction. On the other hand the tocopherol isomers found in the rainbow trout were $\alpha$ and $\beta$ -tocopherol, and $\alpha$-tocopherol had a higher degradation rate by the lipoxygenase catalyzed co-oxidation than of $\beta$-tocopherol in the reaction of model system.

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Optimization for Effective Bioproduction of Natural (-)-1-Octen-3-ol by Lipoxygenase and Hydroperoxide Lyase from Agaricus bisporus (Agaricus bisporus의 Lipoxygenase와 Hydroperoxide Lyase를 이용한 천연 (-)-1-Octen-3-o1 생산 공정의 최적화)

  • Kim, Kyoung-Ju;Kim, Yong-Hwi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.899-903
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    • 2005
  • One of the most important volatile aroma compounds responsible for mushroom flavor is 1-octen-3-ol. To meet the demand for natural mushroom flavor, a study was needed for the production of natural chiral specific (-)-1-octen-3-ol that has higher flavor intensity than synthetic chiral mixtures of (+), and (-)-1-octen-3-ol. The biosynthesis of (-)-1-octen-3-ol was achieved by an aerobic oxidation using lipoxygenase (LOX) and hydroperoxide lyase (HPOL) isolated from commercially available mushrooms in Korean market. Safflower oil from Uiseong, Gyeongsangbuk-do, that contains $\geq75\%$ of linoleic acid, was hydrolyzed using lipase. The recovered linoleic acid was biotransformed to stereo-specific 10-hydroperoxy linoleic acid by LOX. 10- hydroperoxy linoleic acid was further cleaved to (-)-1-octen-3-ol by HPOL. A commercial bioprocess for the production of (-)-1-octen-3-ol was developed using a 5-liter jar fermenter with fruiting bodies of Agaricus bisporus harvested from Buyeo, Chungcheongnam-do. The maximum production of (-)-1-octen-3-o1 was achieved at $4^{\circ}C$, pH 6.5 and 800 rpm yielding 748 mg/kg of mushroom.

DentalVibe versus lignocaine hydrochloride 2% gel in pain reduction during inferior alveolar nerve block in children

  • Menni, Alekhya Chowdary;Radhakrishna, Ambati Naga;Prasad, M. Ghanashyam
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.20 no.6
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    • pp.397-402
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    • 2020
  • Background: Inferior alveolar nerve block (IANB) is the most common, painful, and anxiety-provoking procedure involving needle insertion for anesthetic solution deposition. DentalVibeⓇ (DV) delivers vibration at a sustained frequency as a counter-stimulation to the site of injection, thereby alleviating pain. The aim of this study was to evaluate and compare the effectiveness of DV and lignocaine hydrochloride 2% gel (Lox 2% jelly) in pain reduction during IANB in children. Methods: A split-mouth randomized clinical trial was designed with a sample of 60 children (age, 6 to 12 years) requiring bilateral IANB for various dental procedures; DV was used while administering IANB and Lox 2% jelly was used as the topical anesthetic before administering IANB at subsequent appointments. During both appointments, pain perception was measured using the sound, eye, motor (SEM) scale and Wong-Baker faces pain rating scale (WBFPRS); oxygen saturation (SpO2) and pulse rate were measured using a pulse oximeter before, during, and after the IANB procedure. The obtained values were tabulated and subjected to statistical analysis. Wilcoxon test was used for intergroup comparison, and Friedman test, for intragroup comparison of measured variables at different treatment phases. Results: The medians and interquartile ranges of the WBFPRS scores recorded during the IANB procedure for DV and Lox 2% jelly were 2 (2-4) and 2 (0-2), respectively (P < 0.05). The SEM scale scores, mean SpO2, and pulse rate did not show any significant differences during the IANB procedure between both treatments. Conclusion: Both DV and Lox 2% jelly were found to be effective in pain reduction during IANB in children.

Identification of microRNAs and their target genes in the placenta as biomarkers of inflammation

  • Jang, Hee Yeon;Lim, Seung Mook;Lee, Hyun Jung;Hong, Joon-Seok;Kim, Gi Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.1
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    • pp.42-53
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    • 2020
  • Objective: Recently, microRNA (miRNA) has been identified both as a powerful regulator involved in various biological processes through the regulation of numerous genes and as an effective biomarker for the prediction and diagnosis of various disease states. The objective of this study was to identify and validate miRNAs and their target genes involved in inflammation in placental tissue. Methods: Microarrays were utilized to obtain miRNA and gene expression profiles from placentas with or without inflammation obtained from nine normal pregnant women and 10 preterm labor patients. Quantitative real-time polymerase chain reaction and Western blots were performed to validate the miRNAs and differentially-expressed genes in the placentas with inflammation. Correlations between miRNA and target gene expression were confirmed by luciferase assays in HTR-8/SVneo cells. Results: We identified and validated miRNAs and their target genes that were differentially expressed in placentas with inflammation. We also demonstrated that several miRNAs (miR-371a-5p, miR-3065-3p, miR-519b-3p, and miR-373-3p) directly targeted their target genes (LEF1, LOX, ITGB4, and CD44). However, some miRNAs and their direct target genes showed no correlation in tissue samples. Interestingly, miR-373-3p and miR-3065-3p were markedly regulated by lipopolysaccharide (LPS) treatment, although the expression of their direct targets CD44 and LOX was not altered by LPS treatment. Conclusion: These results provide candidate miRNAs and their target genes that could be used as placental biomarkers of inflammation. These candidates may be useful for further miRNA-based biomarker development.

Evaluation of Achyranthes japonica Ethanol Extraction on the Inhibition Effect of Hyluronidase and Lipoxygenase (쇠무릎 에탄올 추출물의 DPPH, 히알루로니다아제 및 리폭시게나아제 저해 효과)

  • Cho, Kyung-Soon
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1370-1376
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    • 2015
  • The 1, 1- diphenyl 2-picrylhyorazyl (DPPH) is a well-known radical and a trap (scavenger) for other radicals. Hyaluronidase (HAase) is an enzyme that depolymerizes the polysaccharide hyaluronic acid (HA) in the extracellular matrix of connective tissue. Lipoxygenase (LOX) enzyme was reported to convert the arachidonic, linoleic and other polyunsaturated fatty acid into biologically active metabolites involved in the inflammatory and immune responses. The purpose of the present study is to evaluate plant extracts as sources of natural antioxidants and to examine whether Achyranthes japonica having significant DPPH, HAase and LOX inhibitory activity. The inhibitory effect of HAase by A. japonica was assayed using a Morgan microplate assay. The antioxidant activity of the A. japonica extracts was measured on the basis of the scavenging activity of the stable 1, 1- diphenyl 2-picrylhyorazyl (DPPH) free radical. DPPH scavenging activity of matured roots of A. japonica was evaluated at 4.0 mg/ml was 87.8% and that of young roots was 86.2% at same concentration. The roots of A. japonica showed maximum inhibition of HAase activity (IC50 = 27.7 μg/ml). The highest LOX inhibition was recorded in the root extract among three vegetative parts. Inhibition of HAase activity of roots may contribute towards the development of herbal medicines. Although percent inhibition of lipoxygenase by Achyranthes japonica for all young and matured groups for leaves, stems, and roots at different concentrations, there were not show a statistically significant difference (p<0.05).

Characteristics of Lipoxygenase in Black Rice (진도산 흑미의 lipoxygenase의 특성)

  • 이유석;송선주;이종욱
    • Food Science and Preservation
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    • v.6 no.2
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    • pp.216-220
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    • 1999
  • Lipoxygenase(LOX) activity of black rice(Chindo) was measured by spectrophotometric method at In m. Studies at different pH levels revealed that the optimal activity was exhibited at pH 7.0 with 24.97 unit/mg. Enzyme activity was tested at different concentration of the substrate. The apparent Vmax and Km values were determined from the Lineweaver-Burk plot to be 53.85 unit/mg and 0.21 mM. Enzyme activity due to storage temperature (-40, 4 and 25$^{\circ}C$) and period were decreased at all storage temperature. LOX activity of black rice was significantly decreased during the microwave heating.

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ATP-Induced Histamine Release Is in Part Related to Phospholipase $A_2$-Mediated Arachidonic Acid Metabolism in Rat Peritoneal Mast Cells

  • Lee, Yun-Hye;Lee, Seung-Jun;Seo, Moo-Hyun;Kim, Chang-Jong;Sim, Sang-Soo
    • Archives of Pharmacal Research
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    • v.24 no.6
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    • pp.552-556
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    • 2001
  • Histamine and arachidonic acid (AA) release was measured using the P2-purinoceptor antaongists, phospholipase $A_2{\;}(PLA_2)$ and cyclooxygenase (COX)/lipoxygenase (LOX) inhibitors to determine whether or not ATP-induced histamine release is associated with arachidonic acid (AA) release in rat peritoneal mast cells. ATP increased histamine release in a dose dependent manner, whereas adenosine did not. PPADS (a selective P2X-purinoceptor antagonist) and suramin (a nonselective P2X,2Y-purinoceptor antagonist) inhibited ATP-induced histamine release in a dose dependent manner. However, RB-2 (a P2Y-purinoceptor antagonist) did not block ATP-induced histamine release. Manoalide and oleyloxyethyl phosphorylcholine (OPC), secretory PLA$_2$ inhibitors, also inhibited ATP-induced histamine release dose-dependently. Both COX inhibitors (ibuprofen and indomethacin) and LOX inhibitors (baicalein and caffeic acid) inhibited ATP-induced histamine in a dose dependent manner. ATP significantly increased [$^3H$]AA release by 54%. PPADS and suramin significantly inhibited ATP-induced [3H]Ph release by 81% and 39%, respectively. ATP-induced histamine release was significantly inhibited by a variety of protein kinase inhibitors, such as bisindolmaleimide, genistein, methyl 2,5-dihydroxycinnamate, W-7 and trifluoperazine. Overall, the results suggest that ATP-induced histamine release is in part related to the PLA2-mediated AA metabolism and P2X-purinoceptors.

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Effect of Lipoxygenase on the Oxidation of Rainbow Trout Lipid in Model system (모델시스템에 있어서 무지개 송어 지방질의 산화에 대한 Lipoxygenase의 영향)

  • Kim, Hae-Gyoung;Um, Su-Hyon;Cheigh, Hong-Sik
    • Journal of Life Science
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    • v.5 no.2
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    • pp.14-14
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    • 1995
  • The effect of lipoxygenase (LOX) on the oxidation and co-oxidation of lipid fraction was studied in the model system of rainbow trout. For the reaction in model system 1 g of lipid fraction and 50mL of enzyme extract(LOX, 140 unit in 50mL phosphate buffer solution at pH 7,4)), which were obtained from rainbow trout, were homoginized in the presence of Tween 20 and kept at 23$\circ$C for 3 days. The activity of LOX was decreased to 43% of initial level during the reaction in the model system. The initial composition of rainbow trout lipid was showed to be consisted of trigliceride(TG;82%) and free fatty acid(FFA;0.1%), while this converted to 59% of TG and 20% of FIFA, respectively after reaction in model system. Change of fatty acid composition was also observed and the content of linoleic acid, one of the major fatte acids, was decreased to 13% from 54% in the content of total fatty acids after reaction. The carotenoids in rainbow trout were composed of 0.4% $\alpha$-carotene, 1.6% $\beta$ -carotene, 80% canthaxanthin, 7% lutein and 11% zeaxanthin, thus the canthaxanthin was the major component. This canthaxanthin was the most degraded carotenoid by lipoxygenase catalyzed co-oxidation during the reaction. On the other hand the tocopherol isomers found in the rainbow trout were $\alpha$ and $\beta$ -tocopherol, and $\alpha$-tocopherol had a higher degradation rate by the lipoxygenase catalyzed co-oxidation than of $\beta$-tocopherol in the reaction of model system.

Plant growth promoting rhizobacteria influence potato tuberization through enhancing lipoxygenase activity

  • Akula, Nookaraju;Upadhyaya, Chandrama P.;Kim, Doo-Hwan;Chun, Se-Chul;Park, Se-Won
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.18-18
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    • 2010
  • Molecular insights on the role of plant growth promoting rhizobacteria (PGPR) in potato tuberization are reported in the present study. The PGPRwere isolated from the soil collected from potato fields of Highland Agricultural Research Centre, Pyeongchang, Korea and they were identified to the genus level based on the 16S rRNA sequence analysis. These PGPR were heat-killed, filtered and the filtrates were addedindividually at a concentration of $10^7\;cfu\;mL^{-1}$ in MS (Murashige and Skoog's) medium supplemented with 7% (w/v) sucrose to study their influence on in vitro potato tuberization. Tuber initiation occurred early in untreated control, while tuber growth was pronounced in case of PGPR treatments. The control explants showed tuber formation as a result of sub-apical swelling of stolons while several sessile tubers formed directly in the axils of nodal cuttings in case of PGPR treatments, which is an indication of strong induction for tuberization. Theexplants cultured on MS medium supplemented with bacterial isolate 6 (Bacillus firmus strain 40) showed highest average tuber yield (Ca. 12.56 g per treatment) after 30 days of culture, which was 3 folds increase over the untreated control. A significant increase in lipoxygenase (LOX1) mRNA expression and activity of LOX enzyme were also detected in the tubers induced on PGPR treatments as compared to untreated control. This LOX expression level correlated with increased tuber growth and tuber yield. Further studies focused on the role of bacteria cell wall components, growth regulators and signal molecules released by PGPR are under investigation to elicit clues for PGPR-mediated signal pathway controlling potato tuberization.

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Comparison of Methods for Stable Simultaneous Expression of Various Heterologous Genes in Saccharomyces cerevisiae (출아효모에서 다양한 이종 유전자의 안정적 동시발현을 위한 방법의 비교)

  • Jung, Heo-Myung;Kim, Yeon-Hee
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.667-672
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    • 2019
  • We compared two integration systems for stable expression of heterologous genes in Saccharomyces cerevisiae. A Candida glabrata-derived gene was used as the selective marker for the Cre/loxP system, and XYLP, XYLB, GRE3, and XYL2 genes were used as model heterologous genes and ligated into the universal pRS-CMT vector. The resulting pRS-XylP, pRS-XylB, pRS-Gre3, and pRS-Xyl2 plasmids were sequentially integrated into yeast chromosome VII by four integration processes (marker rescue and gene integration). The four introduced genes were successfully expressed. Further, the pRS-PBG2 plasmid harboring expression cassettes for the four genes was constructed for one-step integration. The four genes that were introduced were stably maintained as a gene cluster and were simultaneously expressed. The one-step integration was more effective for the simultaneous integration and expression of the four genes related to xylan/xylose metabolism. This method will enable the generation of a useful biosystem through appropriate use of gene integration methods.