• Title/Summary/Keyword: Xa1

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Epigenetic Study of XIST Gene from Female and Male Cells by Pyrosequencing (남성과 여성에서 XIST 유전자의 후성학적 비교 연구)

  • Kim, Hwan-Hee;Yun, Yeo-Jin;Song, Min-Ae;Lee, Su-Man
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.25-31
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    • 2010
  • Objective: X inactivation is the silencing one of the two X chromosomes in female mammals for gene dosage on the X-chromosome between female and male. X inactivation is controlled by X inactive-specific transcript (XIST) gene, untranslated RNA. XIST is expressed only from the inactive X (Xi), not expressed from the active X (Xa). The Xist promoter is methylated on the silent Xist allele on the Xa in somatic cells, and less methylated on the Xist-expressing Xi. We investigated the difference of XIST methylation pattern of the promoter and 5'-region of XIST from male (XY) and female (XX) subjects. Methods: The direct quantification of XIST methylation is required for clinical application of normal XX and XY blood. Methylation percentage of eight CpG sites (-1696, -1679, -1475, -1473, -1469, +947, +956, +971) of XIST gene were diagnosed by pyrosequencing. Results: We directly quantitated the methylation percentage of the promoter and 5'-end of XIST by pyrosequencing. The average methylation percentages at CpG6-8 sites (+947, +956, +971) were 45.2% at CpG6, 49.9% at CpG7, and 44.2% at CpG8 from normal female and normal male were 90.6%, 96.7%, 87.8%, respectively. Nether CpG 1-5sites (-1696, -1679, -1475, -1473, -1469) had any effect on XX and XY. Conclusion: This method is sensitive for quantifying the small percentage change in the methylation status of XIST, and may be used for diagnosis.

Expression of Antibacterial Protein, Nuecin, Using Baculorivus Expression Vector System in Bm5 Insect Cell and Bombyx mori (누에 배양세포(Bm5) 및 생체에서 베큘로바이러스 발현계를 이용한 누에신 단백질 발현 특성)

  • 윤은영;구태원;황재삼;김상현;강석우;김근영;진병래
    • Journal of Sericultural and Entomological Science
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    • v.44 no.2
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    • pp.69-73
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    • 2002
  • For the practical use of nuecin protein, we tried to overexpress nuecin using Bm5 insect cell and Bombyx mori. We inserted nuecin cDNA into pBm10po1-Xa vector derived from B. mori nuclear polyhedrosis virus (BmNPV), and expressed in Bm5 cells and B. mori respectively. SDS-PAGE and Northern blot analysis showed an expressed of the protein when baculovirus expression vector system (BEVS) was used. The amount of intracellular protein is abundant, but the amount of extracellular protein is poor. The results suggest that the biologically active nuecin protein produced by using BEVS is poor because incresed level of misfolded nuecin by the strong promoter, polyhedrin and p 10 of BEVS, decrease the level of free chaperons and foldases by binding with them.

Characterization of the Pediocin Operon of Pediococcus acidilactici K10 and Expression of His-Tagged Recombinant Pediocin PA-1 in Escherichia coli

  • MOON GI SEONG;PYUN YU RYANG;KIM WANG JUNE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.403-411
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    • 2005
  • The relationship between plasmid (~9.5 kb) and pediocin PA-1 in P. acidilactici K10 was confirmed by plasmid curing. The pediocin operon of P. acidilactici K10 was amplified by PCR (polymerase chain reaction), and the nucleotide sequence was analyzed. The sequence of the pediocin operon of P. acidilactici K10 was similar to those of P. acidilactici strains producing pediocin PA-1/ AcH. For the expression of pediocin PA-1 in E. coli, a pQEPED (pQE-30 Xa::mature pedA) was constructed. His-tagged recombinant pediocin PA-1 (-6.5 kDa) was translated by cell-free in vitro transcription and translation using pQEPED as a DNA template. Theresult of slot blotting assay showed that transcription of recombinant pedA in E. coli M15 was induced by the addition of isopropyl-$\beta$-D-thiogalactopyranoside (IPTG) at the final concentration of 1 mM. Although the recombinant pediocin PA-1 inhibited the growth of E. coli, it was expressed in the host strain and purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography under denaturing condition. This is the first report for the production and one-step purification of biologically active recombinant pediocin PA-1 in E. coli.

Changes in Contents and Composition of Dietary Fiber during Buckwheat Germination (메밀 발아 중 식이섬유 함량과 조성의 변화)

  • 이명헌;우순자
    • The Korean Journal of Food And Nutrition
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    • v.7 no.4
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    • pp.274-283
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    • 1994
  • To provide the efficient application scheme of buckwheat dietary fiber and basic information of seed germination, buckwheat(Fagopyrum esculentum Moench) was germinated at 10$^{\circ}C$ for 7 days and the contents and composition of the total dietary fiber(TDF), insoluble dietary fiber(IDF), soluble dietary fiber(SDF) wire examined at 24 hour intervals. The TDF content in ungerminated seeds was 24.86o on dry weight basis. It decreased for the 1st day of germination, but gradually increased for 7 days afterwords. The contents of IDF and SDF in ungerminated seeds were 22.05, 1.42% respectively. The IDF and SDF contents decreased in the initial stage of germination, but then gradually increased. The composition of the IDF and SDF in the TDF during the germination period showed different tendencies. The IDF decreased with germination time until 5 days and then increased. The SDV increased until 5 days and then decreased gradually. The TDF contents obtained by AOAC method were generally higher than those obtained by Prosky method. The TDF contents obtained by the two method, however, were very closely correlated (r=0.9966, p< 0.01) The IDF(X1) and SDF(X2) showed the significant regression equation(p<0.01) with the root length(Y). The equation was Y: -12.6681+0.5089${\times}$ 1 $\div$ 0.6022Xa and R2 was 0.968.

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Deciphering the Role of Tyrosine Sulfation in Xanthomonas oryzae pv. oryzae Using Shotgun Proteomic Analysis

  • Park, Hye-Jee;Park, Chang-Jin;Bae, Nahee;Han, Sang-Wook
    • The Plant Pathology Journal
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    • v.32 no.3
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    • pp.266-272
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    • 2016
  • A bacterial tyrosine sulfotransferase, RaxST, is required for activation of rice XA21-mediated immunity, and it catalyzes sulfation of tyrosine residues of Omp1X and RaxX in Xanthomonas oryzae pv. oryzae, a causal agent of bacterial blight in rice. Although RaxST is biochemically well-characterized, biological functions of tyrosine sulfation have not been fully elucidated. We compared protein expression patterns between the wildtype and a raxST knockout mutant using shotgun proteomic analysis. Forty nine proteins displayed a more than 1.5-fold difference in their expression between the wildtype and the mutant strains. Clusters of orthologous groups analysis revealed that proteins involved in cell motility were most abundant, and phenotypic observation also showed that the twitching motility of the mutant was dramatically changed. These results indicate that tyrosine sulfation by RaxST is essential for Xoo movement, and they provide new insights into the biological roles of RaxST in cellular processes.

Construction of a Novel Recombinant Bombyx mori Nuclear Polyhedrosis Virus Producing the Fluorescent Polyhedra

  • Kang, Seok-Woo;Yun, Eun-Young;Woo, Soo-Dong;Goo, Tae-Won;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.3 no.1
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    • pp.75-81
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    • 2001
  • We have constructed a novel recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) producing the green fluorescent polyhedra. For the production of the fluorescent polyhedra, partial polyhedrin gene containing KRKK as nuclear localization site from the BmNPV polyhedrin gene and the green fluorescent protein (gfp) gene were introduced under the control of p10 promoter of BmNPV. The recombinant BmNPV was stably produced fluorescent polyhedra in the infected Bm5 cells and the morphology of the fluorescent polyhedra was similar to that of wild-type BmNPV. The fluorescent polyhedra had 32 kDa native polyhedrin and 41 kDa fusion protein. From these data, we have further developed a novel BmNPV p10-based transfer vector producing recombinant polyhedra with foreign gene Product. The novel BmNPV P10-based transfer vector is composed of partial polyhedrin gene, factor Xa, and multiple cloning sites.

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Expression of an Antimicrobial Peptide Magainin by a Promoter Inversion System

  • Lee, Jae-Hyun;Hong, Seung-Suh;Kim, Sun-Chang
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.34-41
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    • 1998
  • A method was developed for the controlled expression of an antimicrobial peptide magainin in Escherichia coli. A series of concatemeric magainin genes was constructed with a gene amplification vector, and fused to the 3'end of malE gene encoding the affinity ligand, E. coli maltose-binding protein (MBP). The construct directed the synthesis of the fusion protein with the magainin polypeptide fused to the C-terminus of MBP. The fusion protein was expressed in a tightly regulatable expression system which was under the control of an invertible promoter. The MBP-fused magainin monomer was expressed efficiently. However, the expression level of the MBP-fused magainin in E. coli decreased with the increasing size of multimers possibly because of the transcription and translation inhibition by the multimeric peptides. After purification using an amylose affinity column, the fusion protein was digested by factor Xa at a specific cleavage site between the monomers. The recombinant magainin had an antimicrobial activity identical to that of synthetic magainin. This experiment shows that a biologically active, antimicrobial peptide magainin can be produced by fusing to MBP, along with a promoter inversion vector system.

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Enoxaparin as an Anticoagulant in a Multipara with a Mechanical Mitral Valve: A Case Report

  • Yo Seb Lee;Jun Seok Kim
    • Journal of Chest Surgery
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    • v.56 no.6
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    • pp.452-455
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    • 2023
  • Patients who have undergone mechanical valve replacement require anticoagulation therapy with warfarin to prevent thromboembolism. However, administering warfarin to pregnant patients increases their risk of warfarin embryopathy or central nervous system disorders. Consequently, safer alternatives, such as heparin or low-molecular-weight heparin injection, are substituted for warfarin. However, limited research has been conducted on this subject, with no large-scale studies and particularly few investigations involving multiparous patients. A patient who had previously undergone mechanical mitral valve replacement for atrial septal defect and mitral stenosis received anticoagulant therapy with enoxaparin during 2 pregnancies. Upon confirmation of pregnancy, warfarin was replaced with subcutaneously injected enoxaparin with a dosage of 1 mg/kg at 12-hour intervals. The enoxaparin dosage was controlled using an anti-factor Xa assay, with a target range of 0.3-0.7 IU/mL. Intravenous heparin injections were administered starting 3 days prior to the expected delivery date and were continued until delivery, after which warfarin was resumed. No complications were observed during the deliveries.

Syntheses and realization of Quaternary Galois Field Sum-Of-Product(QGFSOP) expressed 1-variable functions Permutational Literals (치환리터럴에 의한 Quaternary Galois Field Sum-Of-Product(QGFSOP)형 1-변수 함수의 합성과 실현)

  • Park, Dong-Young;Kim, Baek-Ki;Seong, Hyeun-Kyeong
    • Journal of Advanced Navigation Technology
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    • v.14 no.5
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    • pp.710-717
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    • 2010
  • Even though there are 256 possible 1-qudit(1-variable quantum digit) functions in quaternary logic, the most useful functions are 4!=24 ones capable of representing in QGFSOP expressions by possible permuting of 0,1,2, and 3. In this paper, we propose a permutational literal(PL) representation and a QPL(Quaternary PL) gate which use the operands of a multiplicand A and an augend D in $Ax^C$+D(GF4) operation as a control variable of multi-cascaded PLs. And we also present new PL synthesis algorithms to synthesize QGFSOP expressed 24 (1-qudit) functions by applying three PL operators as ab(mutual permutation), + D(addition), and XA (multiplication). Finally architectures, circuits, and a CMOS implementation to realize proposed PL synthesis algorithms for $Ax^C$+D(GF4) functions are presented.

Comparisons of Bleeding Risks between Rivaroxaban and Dalteparin for Treatment of Venous Thromboembolism in Cancer Patients (암환자의 정맥혈전색전증 치료에서 rivaroxaban과 dalteparin의 출혈 부작용 비교)

  • Kim, Yoon Kyung;An, Sook Hee;Kim, Jae Yeon;Chung, Jee Eun;Gwak, Hye Sun
    • Korean Journal of Clinical Pharmacy
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    • v.26 no.3
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    • pp.195-200
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    • 2016
  • Background: Venous thromboembolism (VTE) is a common and life-threating condition in cancer patients. Low molecular weight heparins (LMWH), such as dalteparin, are recommended in the treatment of VTE. Also, rivaroxaban, an orally administered direct factor Xa inhibitor, was approved for the treatment of VTE. It showed similar efficacy to standard therapy (LMWH or warfarin) and was associated with significantly lower rates of major bleedings. However, in the real world, bleeding has been reported to occur frequently in cancer patient receiving rivaroxaban. The goal of this research was to analyze bleeding risks between rivaroxaban and dalteparin for treatment of VTE in cancer patients. Methods: Medical records of oncology patients who were treated with rivaroxaban or dalteparin for VTE from July 2012 to June 2014 were retrospectively reviewed. Data collected were as follows: age, sex, weight, height, cancer types and stages, ECOG (eastern cooperative oncology group) PS (performance score), VTE types, concurrently used medications, study drug information (dose and duration of therapy), INR (international normalized ratio), PT (prothrombin time), and platelet counts. Bleeding was classified into major bleedings, clinically relevant non-major bleedings, and minor bleedings. Results: A total of 399 patients were included in the study. Of these patients, 246 were treated with rivaroxaban and 153 with dalteparin. Bleeding rates were significantly higher in the rivaroxaban group than in the dalteparin group (adjusted odds ratio (AOR) 2.09, 95% CI 1.22-3.60) after adjusting for confounders. In addition, rivaroxaban remained independently associated with 1.78-fold (95% CI 1.14-2.76) shorter time to bleeding compared to dalteparin after adjusting other factors known to be associated with poor outcomes. Conclusion: This study suggested that rivaroxaban was associated with an increased risk of bleedings in cancer patients.