• Title/Summary/Keyword: fab E

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RNase P-dependent Cleavage of Polycistronic mRNAs within Their Downstream Coding Regions in Escherichia coli

  • Lee, Jung-Min;Kim, Yool;Hong, Soon-Kang;Lee, Young-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.29 no.6
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    • pp.1137-1140
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    • 2008
  • M1 RNA, the catalytic subunit of Escherichia coli RNase P, is an essential ribozyme that processes the 5' leader sequence of tRNA precursors (ptRNAs). Using KS2003, an E. coli strain generating only low levels of M1 RNA, which showed growth defects, we examined whether M1 RNA is involved in polycistronic mRNA processing or degradation. Microarray analysis of total RNA from KS2003 revealed six polycistronic operon mRNAs (acpP-fabF, cysDNC, flgAMN, lepAB, phoPQ, and puuCBE) showing large differences in expression between the adjacent genes in the same mRNA transcript compared with the KS2001 wild type strain. Model substrates spanning an adjacent pair of genes for each polycistronic mRNA were tested for RNase P cleavage in vitro. Five model RNAs (cysNC, flgMN, lepAB, phoPQ, and puuBE) were cleaved by RNase P holoenzyme but not by M1 RNA alone. However, the cleavages occurred at non-ptRNA-like cleavage sites, with much less efficiency than the cleavage of ptRNA. Since cleavage products generated by RNase P from a polycistronic mRNA can have different in vivo stabilities, our results suggest that RNase P cleavage may lead to differential expression of each cistron.

Isolation of an Anticariogenic Compound from Magnoliae Bark (후박피(Magnoliae bark)로부터 항충치활성을 갖는 물질의 분리)

  • Lee, Youn-Soo;Park, Hun-Joo;You, Jae-Sun;Park, Hyung-Hwan;Kwon, Ik-Boo;Lee, Hyeon-Yong
    • Korean Journal of Food Science and Technology
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    • v.30 no.1
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    • pp.230-236
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    • 1998
  • We have screened total 32 herbal drugs to find the highest activity against human cariogenic enzyme, glucosyltransferase (GTase) from the extracts of Magnoliae bark. The extracts were separated into three phases, i. e. water, n-butanol and ethylacetate according to their solvent polarity. Among them, ethylacetate fraction had approximately more than 70% of total activities, and the active principle was further isolated by prep. HPLC following silicagel column chromatography to yield single compound as white powder. The chemical structure of the compound was finally elucidated to be 4,4'-dihydroxy-3,3'-dimethoxylignan from the spectral data of FAB-MS. $^1H-\;and\;^{13}C-NMR$ spectrometries. The compound was also shown to have relatively strong antibacterial activity against ten types of cariogenic oral bacteria and one kind of Actinomyces sp.

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An Antifungal Antibiotic Purified from Bacillus megaterium KL39, a Biocontrol Agent of Red-Pepper Phytophthora-Blight Disease

  • JUNG HEE KYOUNG;KIM SANG-DAL
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1001-1010
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    • 2005
  • Bacillus megaterium KL39, an antibiotic-producing plant growth promoting rhizobacterium (PGPR), was selected from soil. The antifungal antibiotic, denoted KL39, was purified from culture filtrate by column chromatography using Dion HP-20, Silica gel, Sephadex LH-20, and prep-HPLC. Thin layer chromatography, employing the solvent system of ethanol:ammonia:water=8:1:1, showed the $R_{f}$. value of 0.32. The antibiotic KL39 showed a negative reaction with ninhydrin solution, positive with iodine vapor, and also positive with Ehrlich reagent. It was soluble in methanol, ethanol, butanol, and acetonitrile, but insoluble in chloroform, toluene, hexane, ethyl ether, or acetone. Its UV spectrum had the maximum absorption at 208 nm. Amino acid composition, FAB-mass, $^{1}H-NMR,\;^{13}C-NMR$, and atomic analyses showed that the antibiotic KL39 (MW=1,071) has a structure very similar to iturin E. The antibiotic KL39 has a broad antifungal spectrum against a variety of plant pathogenic fungi including Rhizoctonia solani, Pyricularia oryzae, Monilinia froeticola, Botrytis cinenea, Altenaria kikuchiana, Fusarium oxysporum, and F. solani. An MIC value of $10\;{\mu}g/ml$ was determined for Phytophthora capsici. Macromolecular incorporation studies with P. capsici using radioactive [$^{3}H-adenine$] as the precursor, indicated that the antibiotic KL39 strongly inhibits the DNA biosynthesis of the fungal cell. Microscopic observation of the antifungal action showed abnormal hyphal swelling of P. capsici. The purified antibiotic KL39 was very effective for the biocontrol of in vivo Phytophthora-blight disease of pepper.

Prognostic Value of a CYP2B6 Gene Polymorphism in Patients with Acute Myeloid Leukemia

  • Alazhary, Nevin M;Shafik, Roxan E;Shafik, Hanan E;Kamel, Mahmoud M
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.11
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    • pp.4583-4587
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    • 2015
  • Background: The objectives of this study aimed to detect a CYP2B6 polymorphism in de novo cases of acute myeloid leukemia patients and identify any role in disease progression and outcome. Materials and Methods: DNA was isolated from peripheral blood of 82 newly diagnosed acute myeloid leukemia cases and the CYP2B6 G15631T gene polymorphism was assayed by PCR restriction fragment length polymorphism (PCR-RFLP). Results: The frequency of the GG genotype (wild type) was 48 (58.5%) and that of the mutant type T allele was 34 (41.9%). GT genotype heterozygous variants were found in 28 (34%), and TT genotype homozygous variants in 6 (7.3%) cases. We found no significant association between the CYP2B6 G15631T polymorphism and complete response (CR) (p-value=0.768), FAB classification (p-value=0.51), cytogenetic analysis (p-value=0.673), and overall survival (p-value=0.325). Also, there were no significant links with early toxic death (p-value=0.92) or progression-free survival (PFS) (p-value=0.245). Conclusions: Our results suggest that the CYP2B6 polymorphism has no role in disease progression, therapeutic outcome, patient free survival, early toxic death and overall survival in acute myeloid leukemia patients.

A human monoclonal antibody $F_{ab}$ reactive to oxidized LDL and carbamylated LDL recognizes human and mouse atherosclerotic lesions

  • Jang, Young-Ju;Joo, Hee-Jae;Yang, Jeong-In;Seo, Chang-Won;Chung, Kui-Yea;Lanza, Gregory M.;Zhang, Huiying
    • Animal cells and systems
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    • v.15 no.4
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    • pp.259-267
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    • 2011
  • This study was undertaken to produce a $F_{ab}$ fragment of a human monoclonal antibody reactive to oxidized and carbamylated low-density lipoprotein (oxLDL and cLDL) using phage display technology. An analysis of DNA sequences of this $F_{ab}$, termed plaque 15,16-46 $F_{ab}$, revealed that the rearranged $V_H$ was highly mutated. Complementarity-determining regions of the $V_H$ showed a very high R/S ratio and contained many positively charged amino acids. In direct binding and competitive ELISA, the $F_{ab}$ reacted strongly with both MDA-LDL and Cu-oxLDL forms of oxLDL, and also showed high affinity for cLDL. Immunofluorescence and immunohistochemical analyses showed that this $F_{ab}$ positively stained atherosclerotic aortic plaques in $ApoE^{-/-}$ mice as well as those in patients with atherosclerosis. The $F_{ab}$ also showed positive staining in placental decidua from patients with preeclampsia. It is suggested that the plaque 15,16-46 $F_{ab}$ against oxLDL and cLDL might possibly be applicable for developing a diagnostic reagent for both human and rodent animal research to detect and characterize atherosclerotic disease progression in atherosclerotic lesions as well as exploring the pathogenesis of atherogenic diseases such as preeclampsia.

Clinical Impact of Overexpression of FOXP3 and WT1 on Disease Outcome in Egyptian Acute Myeloid Leukemia Patients

  • Assem, Magda M;Osman, Ahmed;Kandeel, Eman Z;Elshimy, Reham AA;Nassar, Hanan R;Ali, Radwa E
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.10
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    • pp.4699-4711
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    • 2016
  • Background: In the last decade, it has become clear that change of gene expression may alter the hematopoietic cell quiescent state and consequently play a major role in leukemogenesis. WT1 is known to be a player in acute myeloid leukemia (AML) and FOXP3 has a crucial role in regulating the immune response. Objectives: To evaluate the impact of overexpression of WT1and FOXP3 genes on clinical course in adult and pediatric AML patients in Egypt. Patients and methods: Bone marrow and peripheral blood samples were obtained from 97 de novo non M3 AML patients (63 adult and 34 pediatric). Real-time quantitative PCR was used to detect overexpression WT1 and FOXP3 genes. Patient follow up ranged from 0.2 to 39.0 months with a median of 5 months. Results: In the pediatric group; WT1 was significantly expressed with a high total leukocyte count median 50X109/L (p=0.018). In the adult group, WT1 had an adverse impact on complete remission induction, disease-free survival and overall survival (p=0.02, p=0.035, p=0.019 respectively). FOXP3 overexpression was associated with FAB subtypes AML M0 +M1 vs. M2, M4+M5 (p =0.039) and the presence of hepatomegaly (p=0.005). Conclusions: WT1 and FOXP3 overexpression has an adverse impact on clinical presentation, treatment response and survival of pediatric and adult Egyptian AML patients.

Isolation and Some Properties of Bitter Taste Compounds from Cultured Oyster, Crassostrea gigas (양식산 굴로부터 쓴맛 성분의 분리 및 성질)

  • LEE Jong-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.1
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    • pp.98-104
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    • 1995
  • Five bitter taste compounds (OY-22, OY-23, OY-24, OY-25 and OY-26) were firstly isolated from cultured oyster (Crassostrea gigas) at Gamak Bay, Sourthern coast of Korea, between November, 1989 and January, 1990, and smoked-canned oyster, which were produced by the same oysters. They were presumed as cyclic peptides composed with 6 or 7 amino acids, including sulfur on the basis of NMR and MS spectra. Val and Leu in OY-24, leu and lie in OY-25 and tow leucines in OY-26 were detected from those each compounds, seperately, by the amino acid analysis. Another amino acids were thought as non-constitutional amino acids. They showed non-toxic to mice $(100{\mu}g/20g\;mice i.p.)$ and non actibacterial artivities to Asp. niger and B. subtilis $(10{\mu}g/disk)$. The chemical structures and other biological activities of them are now in studying.

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Ergosterol peroxides from the fruit body of Sparassis crispa (꽃송이버섯(Sparassis crispa) 자실체로부터 ergosteol peroxide의 분리 및 동정)

  • Lee, Yeong-Geun;Thi, Nhan Nguyen;Kim, Hyoung-Geun;Lee, Dae Young;Lee, Seung-Eun;Kim, Geum-Soog;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.313-316
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    • 2016
  • Sparassis crispa fruits were extracted in 80 % MeOH, and the concentrated extract was partitioned into EtOAc, n-butyl alcohol, and water fractions. The repeated octadecyl $SiO_2$ and silica gel ($SiO_2$) column chromatographies for the EtOAc and nbutyl alcohol fractions led to isolation of two ergosterol peroxides. There chemical structures were determined as ($3{\beta}$,$5{\alpha}$,$8{\alpha}$,22E)-5,8-Epidioxyergosta-6,22-dien-3-ol (ergosterol peroxide) (1) and 3-O-${\beta}$-D-glucopyranosyl ergosterol peroxide (2) based on spectroscopic data analyses including nuclear magnetic resonance, infrared spectrometry, and mass spectrometry (MS). Compounds 1 and 2 were for the first time isolated from S. crispa in this study.

The inflammatory activity of purified-ferulic acid from Tetragonia tetragonioides

  • Kim, Na-Hyeon;Park, Hye-Jin;Lee, Eun-Ho;Cho, Eun-Bi;Kang, In-Kyu;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.62 no.3
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    • pp.239-246
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    • 2019
  • In this study, an evaluation of the anti-inflammatory effect of ferulic acid isolated from Tetragonia tetragonioides in lipopolysaccharide (LPS) simulated RAW 264.7 cells was made. The chemical structure of the active compound was elucidated by $^1H$-NMR, $^{13}C$-NMR, and FAB-MS, and was confirmed to be ferulic acid. Ferulic acid was purified via open column chromatography with Sephadex LH-20 and MCI gel CHP-20. To test the anti-inflammatory effect of ferulic acid, LPS-stimulated RAW 264.7 cells were treated in subsequent experiments with different concentrations of ferulic acid (5, 10, and $25{\mu}g/mL$) and the levels of inflammatory cytokines and enzymes were also measured by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. Cell viability was above 95% at acid concentrations ranging from $5-25{\mu}g/mL$. The results showed that 30% of the production of nitric oxide and 66% of prostaglandin $E_2$ were inhibited by $25{\mu}g/mL$ of ferulic acid, it also inhibited the protein expression of both inducible nitric oxide synthase and cyclooxygenase-2 by 70%. Additionally, it inhibited the production of the pro-inflammatory cytokines, tumor necrosis factor-${\alpha}$, interleukin-6, and interleukin-$1{\beta}$ by 40, 75, and 77%, respectively. According to these results, the anti-inflammatory activity of ferulic acid was demonstrated via his implication in the inhibition of the expression and secretion of inflammatory substances in LPS-stimulated RAW 264.7 cells. Therefore, we concluded that ferulic acid can be used as a functional additive having anti-inflammatory activity.

Identification of Maysin and Related Flavonid Analogues in Corn Silks (옥수수 수염에서 Maysin 및 유사물질의 동정)

  • Kim, Sun-Lim;Snook, Maurice E.;Kim, E-Hun;Park, Cheol-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.3
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    • pp.151-157
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    • 2000
  • This study was carried out to isolate and identify the maysin and related flavonoid analogues in corn silks. Silks were covered with silk bag to prevent pollination and were sampled at 3-5 days after silking. The silks were filled with 100% MeOH and stored at $0^{\circ}C$ until analysis. The MeOH extracts of corn silks were filtered and concentrated at 35-4$0^{\circ}C$. The ${CH}_2$${Cl}_2$ was added on the concentrated aqueous solution to remove the chlorophyll and lipids. The Cis open column (25mm$\times$54 cm) was washed and activated with serial treatment of 500$m\ell$ of 100% MeOH(twice)longrightarrow75% MeOH longrightarrow50% MeOHlongrightarrow30% MeOHlongrightarrow100% $H_2$O(2 times). The concentrated aqueous solution was applied to the $C_{18}$ column and washed with $H_2O$ several times to remove the sugars and water soluble pigments. Neochlorogenic acid, chlorogenic acid and 4-caffeoylquinic acid were eluted with 10% MeOH, and rhamosyl isoorientin was eluted with 30% MeOH, but maysin was eluted with 50% MeOH from the $C_18$ open column. Collected fractions were analyzed with HPLC by using revers-phase Ultras-phere $C_{18}$ column (4.6$\times$250mm, 5$\mu\textrm{m}$) and $H_2$O (10% MeOH containing 0.1% $H_3$${PO}_4$)/MeOH (100% MeOH containing 0.1% H$_3$PO$_4$) linear gradient from 20% to 90% MeOH for 35 minutes, a flow rate of 1 $m\ell$/min and detection at 340nm. The selected fractions were concentrated and applied to the silicic acid column. Maysin was eluted with 500$m\ell$ of 100% ethyl acetate from the silicic acid column for the first purification, and the purity of collected fractions was about 75%, but the purity from the second purification with the Cis column (1/2 $\times$ 43") was greater than 95%. FAB-MS spectral data was obtained with VG7O-VSEQ VG analytical fast atom bombardment mass (UK). $^1$H-NMR and $^{13}$ C-NMR data were obtained with Bruker DPX 400 MHz NMR spectrometers (German) in DMSO-d$_{6}$ at 400 and 100 MHz, respectively.vely.

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