• Title/Summary/Keyword: $S_N2$ reaction

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Cloning and Phylogenetic Characterization of Coat Protein Genes of Two Isolates of Apple mosaic virus from ¡?Fuji¡? Apple

  • Lee, Gung-Pyo;Ryu, Ki-Hyun;Kim, Hyun-Ran;Kim, Chung-Sun;Lee, Dong-Woo;Kim, Jeong-Soo;Park, Min-Hye;Noh, Young-Mi;Choi, Sun-Hee;Han, Dong-Hyun;Lee, Chang-Hoo
    • The Plant Pathology Journal
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    • v.18 no.5
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    • pp.259-265
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    • 2002
  • Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was detected and isolated from diseased 'Fuji' apple (Malus domestica) in Korea. The coat protein (CP) genes of two ApMV strains, denoted as ApMV-Kl and ApMV-K2, were amplified by using the reverse transcription and polymerase chain reaction (RT-PCR) and were analyzed thereafter. The objectives were to define the molecular variability of genomic information of ApMV found in Korea and to develop virus-derived resistant gene source for making virus-resistant trans-genic apple. RT-PCR amplicons for the APMVS were cloned and their nucleotide sequences were determined. The CPs of ApMV-Kl and ApMV-K2 consisted of 222 and 232 amino acid residues, respectively. The identities of the CPs of the two Korean APMVS were 93.1% and 85.6% at the nucleotide and amino acid sequences, respectively. The CP of ApMV-Kl showed 46.1-100% and 43.2-100% identities to eight different ApMV strains at the nucleotide and amino acid levels, respectively. When ApMV-PV32 strain was not included in the analysis, ApMV strains shared over 83.0% and 78.6% homologies at the nucleotide and amino acid levels, respectively. ApMV strains showed heterogeneity in CP size and sequence variability. Most of the amino acid residue differences were located at the N-termini of the strains of ApMV, whereas, the middle regions and C-termini were remarkably conserved. The APMVS were 17.(1-54.5% identical with three other species of the genus Ilarviyus. ApMV strains can be classified into three subgroups (subgroups I, II, and III) based on the phylogenetic analysis of CP gene in both nucleotide and amino acid levels. Interestingly, all the strains of subgroup I were isolated from apple plants, while the strains of subgroups II and III were originated from peach, hop, or pear, The results suggest that ApMV strains co-evolved with their host plants, which may have resulted in the CP heterogeneity.

Apoptosis and Apoptosis Related Gene Expression in Preimplantation Porcine Diploid Parthenotes Developing In Vitro (착상전 이배체 단위발생 돼지난자의 체외 배양에서 세포사멸과 세포사멸에 관여하는 유전자의 발현에 관한 연구)

  • X. S. Cui;Kim, I. H.;Kim, N. H.
    • Korean Journal of Animal Reproduction
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    • v.27 no.2
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    • pp.169-177
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    • 2003
  • This study was conducted to determine effects of polyvinyl alcohol (PVA), fetal bovine serum (FBS) bovine serum albumin (BSA) and epidermal growth factor (EGF) on blastocoel formation, total cell number, apoptosis and apoptosis-related gene expression of porcine diploid parthenotes developing in vitro. The addition of 0.4% BSA to the culture medium enhanced the development of 2-cell stage parthenotes to the blastocysts stage (P<0.01). FBS reduced cell numbers of blastocysts (P<0.01) and increased percentage of apoptosis in the blastocysts (P<0.001). However, while BSA increased cell numbers, it did so only when EGF was present. Either agent on its own had no effect. Similarly, apoptosis in the blastocysts was not influenced by either agent on its own but was reduced when both BSA and EGF were present. Furthermore, semi-quantitative reverse-transcriptase polymerase chain reaction revealed that EGF enhanced the mRNA expression of Bcl-xL in the presence of 0.4% BSA but BSA and EGF alone had no effect, and EGF and/or BSA did not influence Bak gene expression in the blastocyst stage parthenotes. However FBS reduced Bcl-xL mRNA expression (P <0.05) and enhanced Bak expression. This result suggests that apoptosis related genes expression is significantly affected by supplements, which may result in alteration of apoptosis and embryo viability of porcine embryos developing in vitro.

Gene cloning of β-mannanase C from Cellulosimicrobium sp. YB-43 and characterization of the enzyme (Cellulosimicrobium sp. YB-43으로부터 mannanase C 유전자의 클로닝과 효소 특성)

  • Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.126-135
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    • 2018
  • The characteristics of enzyme and gene for mannanase B had been reported from Cellulosimicrobium sp. YB-43 producing some kind of mannanase. A gene coding for the enzyme, named mannanase C (ManC), was expected to be located downstream of the manB gene. The manC gene was cloned by polymerase chain reaction and sequenced completely. From this nucleotide sequence, ManC was identified to consist of 448 amino residues and contain a carbohydrate binding domain CBM2 besides a catalytic domain, which was homologous to mannanase belonging to the glycosyl hydrolase family 5. The catalytic domain of ManC showed the highest amino acid sequence similarity of 55% with the mannanases from Streptomyces sp. SirexAA-E (55.8%; 4FK9_A) and S. thermoluteus (57.6%; BAM62868). The His-tagged ManC (HtManC) lacking N-terminal signal peptide with hexahistidine at C-terminus was produced and purified from cell extract of recombinant Escherichia coli. The purified HtManC showed maximal activity at $65^{\circ}C$ and pH 7.5, with no significant change in its activity at pH range from 7.5 to 10. HtManC showed more active on konjac and locust bean gum (LBG) than guar gum and ivory nut mannan (ivory nut). Vmax and Km values of the HtManC for LBG were 68 U/mg and 0.45 mg/ml on the optimal condition, respectively. Mannobiose and mannotriose were observed on TLC as major products resulting from the HtManC hydrolysis of mannooligosacharides. In addition, mannobiose and mannose were commonly detected as the hydrolyzed products of LBG, konjac and ivory nut.

MicroRNA-21 promotes epithelial-mesenchymal transition and migration of human bronchial epithelial cells by targeting poly (ADP-ribose) polymerase-1 and activating PI3K/AKT signaling

  • Zhang, Shiqing;Sun, Peng;Xiao, Xinru;Hu, Yujie;Qian, Yan;Zhang, Qian
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.4
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    • pp.239-253
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    • 2022
  • Epithelial-mesenchymal transition (EMT) is known to be involved in airway remodeling and fibrosis of bronchial asthma. However, the molecular mechanisms leading to EMT have yet to be fully clarified. The current study was designed to reveal the potential mechanism of microRNA-21 (miR-21) and poly (ADP-ribose) polymerase-1 (PARP-1) affecting EMT through the PI3K/AKT signaling pathway. Human bronchial epithelial cells (16HBE cells) were transfected with miR-21 mimics/inhibitors and PARP-1 plasmid/small interfering RNA (siRNA). A dual luciferase reporter assay and biotin-labeled RNA pull-down experiments were conducted to verify the targeting relationship between miR-21 mimics and PARP-1. The migration ability of 16HBE cells was evaluated by Transwell assay. Quantitative real-time polymerase chain reaction and Western blotting experiments were applied to determine the expression of Snail, ZEB1, E-cadherin, N-cadherin, Vimentin, and PARP-1. The effects of the PI3K inhibitor LY294002 on the migration of 16HBE cells and EMT were investigated. Overexpression of miR-21 mimics induced migration and EMT of 16HBE cells, which was significantly inhibited by overexpression of PARP-1. Our findings showed that PARP-1 was a direct target of miR-21, and that miR-21 targeted PARP-1 to promote migration and EMT of 16HBE cells through the PI3K/AKT signaling pathway. Using LY294002 to block PI3K/AKT signaling pathway resulted in a significant reduction in the migration and EMT of 16HBE cells. These results suggest that miR-21 promotes EMT and migration of HBE cells by targeting PARP-1. Additionally, the PI3K/AKT signaling pathway might be involved in this mechanism, which could indicate its usefulness as a therapeutic target for asthma.

A Loop-mediated Isothermal Amplification Method for White-backed Planthopper-specific Detection (고리매개등온증폭법(LAMP)을 이용한 흰등멸구 특이 판별법)

  • Seo, Bo Yoon;Park, Chang Gyu;Jung, Jin Kyo;Cho, Jumrae;Lee, Gwan-Seok;Kim, Kwang-Ho
    • Korean journal of applied entomology
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    • v.57 no.4
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    • pp.393-399
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    • 2018
  • A loop-mediated isothermal amplification (LAMP) primer set (WBPH-65) was designed for the species-specific detection of white-backed planthopper (WBPH) Sogatella furcifera based on the full-length sequence of the internal transcribed spacer 2 (ITS2) (KC417469.1). The WBPH-65 primer set consists of six primers (total 165 bp), F3 (18 bp), B3 (18 bp), FIP (43 bp), BIP (40 bp), LF (21 bp), and LB (25 bp). After the LAMP reaction of three rice planthoppers, S. furcifera, Nilaparvata lugens, and Laodelphax striatellus, with the WBPH-65 primer set for 60 min at $65^{\circ}C$, the LAMP products were observed in the genomic DNA of S. furcifera only. According to the DNA amount of S. furcifera and incubation duration at $65^{\circ}C$, the difference of fluorescence relative to the negative control (0 ng) was clearly observed in a 40-min incubation with 10 and 100 ng or in case of 60-min incubation with 0.01, 0.1, 1, 10, and 100 ng. There was little difference in fluorescence between the negative control and all the other DNAs tested in 20- and 30-min incubations. On the other hand, the WBPH-65 primer set without LF and LB primers showed little amplification in the genomic DNAs of the three rice planthoppers, S. furcifera, N. lugens, and L. striatellus in a 60-min incubation. These results suggest that all six primers (F3, B3, FIP, BIP, LF, and BF) are necessary for the WBPH-65 primer set to detect S. furcifera within a 60-min incubation, and is able to discriminate S. furcifera from at least N. lugens and L. striatellus.

Stability of Anti-Helicobacter pylori IgY Under Some Condition (Anti-Helicobacter pylori IgY의 처리조건에 따른 안정성에 관한 연구)

  • Jung, S.H.;Kim, H.J.;Lee, S.W.;Lee, N.H.
    • Journal of Animal Science and Technology
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    • v.47 no.5
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    • pp.851-856
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    • 2005
  • The aim of this experiment was to evaluate the stability of anti-Helicobacter pylori IgY in water soluble fraction(WSF) of egg yolk according to the heat, pH and digestive enzyme treatment. Anti-H. pylori IgY content of WSF remained 76% after pasteurization(63$^{\circ}C$ for 30 min). The stability of anti-H. pylori IgY at different pH showed a tendency to diminish according to decreasing pH from 7.0 to 1.5(p<0.05). Anti-H. pylori IgY content was 84.4% after treatment for 1 hour at 37$^{\circ}C$ in pH 5.0. There were significantly differences in IgY content between 1 hour and 2 hours at pH 2.0 in 200 units of pepsin treatment(p<0.05). However, IgY was relatively stable at pH 4.0 regardless of the reaction time and the concentration of pepsin. The stability of IgY of egg yolk after the treatment of trypsin was significantly higher than that of water soluble fraction (p<0.05). This results indicated that anti-H. pylori IgY showed relatively a good stability on heat, pH and digestive enzyme.

Cyclosporine A (Cipol-$N^{(R)}$) Therapy in Children with Idiopathic Nephrotic Syndrome (소아 특발성 신증후군에서의 cyclosporine A(Cipol-$N^{(R)}$)의 치료 효과)

  • Hong Ihn Hee;Ko Cheol Woo;Koo Ja Hoon;Kim Ji-Hong;Kim Pyung-Kil;Cho Byoung Soo
    • Childhood Kidney Diseases
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    • v.3 no.1
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    • pp.48-56
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    • 1999
  • Purpose : This multicenter collaboratory study was conducted to see the therapeutic efficacy and side effect of cyclosporine A (Cipol-$N^{(R)}$, Chong Kun Dang) on children with idiopathic nephrotic syndrome who experienced frequently relapsing (FR), steroid dependent (SD), or steroid resistant (SR) pattern. Patients and methods : Thirty-nine children with SD/FR NS and 3 children with SR NS were enrolled in the study. After induction of remission (SD/FR NS) with steroid or after 4 weeks of steroid therapy (SR NS), cyclosporine A was started in a dose of 4-5 mg/Kg/day in two divided dose and steroid (prednisolone or equivalent dose of deflazacort) was tapered slowly. During 16 weeks of study period, monthly check up of physical examination and various laboratory tests including BUN, creatinine, Ccr and cyclosporine blood level were done. Results : Out of 39 children with SD/FR NS, 35($89.7\%$) maintained sustained remission and at 4 weeks after therapy, values of serum protein, albumin, cholesterol, and 24 hours urinary protein excretion showed normal values. Two out of 3 children with SR NS showed and sustained remission with cyclosporine A therapy. Side reaction to cyclosporine A therapy showed hypertrichosis in 8 cases and hyperuricemia in 5 cases. However, other laboratory tests including CBC, liver profile, BUN, creatinine and GFR (creatinine clearance utilizing 24 hour urine) did not show any abnormalities during the 16 weeks of study period. Conclusion : Cyclosporine A (Cipoi-$N^{(R)}$ Chong Kun Dang) can be utilized quite effectively on children with SD/FR or SR NS and further trial of cyclosporine A on long-term basis (1-2 year period) is needed to determine it's efficacy and side effect (especially nephrotoxicity) of long-term administration of cyclosporine A.

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Surface Structures and Thermal Desorption Behaviors of Cyclopentanethiol Self-Assembled Monolayers on Au(111)

  • Kang, Hun-Gu;Kim, You-Young;Park, Tae-Sun;Park, Joon-B.;Ito, Eisuke;Hara, Masahiko;Noh, Jae-Geun
    • Bulletin of the Korean Chemical Society
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    • v.32 no.4
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    • pp.1253-1257
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    • 2011
  • The surface structures, adsorption conditions, and thermal desorption behaviors of cyclopentanethiol (CPT) self-assembled monolayers (SAMs) on Au(111) were investigated by scanning tunneling microscopy (STM), X-ray photoelectron spectroscopy (XPS), and thermal desorption spectroscopy (TDS). STM imaging revealed that although the adsorption of CPT on Au(111) at room temperature generates disordered SAMs, CPT molecules at $50^{\circ}C$ formed well-ordered SAMs with a $(2{\surd}3{\times}{\surd}5)R41^{\circ}$ packing structure. XPS measurements showed that CPT SAMs at room temperature were formed via chemical reactions between the sulfur atoms and gold surfaces. TDS measurements showed two dominant TD peaks for the decomposed fragments ($C_5H_9^+$, m/e = 69) generated via C-S bond cleavage and the parent molecular species ($C_5H_9SH^+$, m/e = 102) derived from a recombination of the chemisorbed thiolates and hydrogen atoms near 440 K. Interestingly, dimerization of sulfur atoms in n-alkanethiol SAMs usually occurs during thermal desorption and the same reaction did not happen for CPT SAMs, which may be due to the steric hindrance of cyclic rings of the CPT molecules. In this study, we demonstrated that the alicyclic ring of organic thiols strongly affected the surface structure and thermal desorption behavior of SAMs, thus providing a good method for controlling chemical and physical properties of organic thiol SAMs.

Isolation and Genetic Study of Hantavirus from Apodemus peninsulae Captured in Yeuncheon-gun, Kyunggi-do (경기도에서 채집한 Apodemus peninsulae에서 한탄바이러스 분리와 유전학적 연구)

  • Song, Ki-Joon;Kim, Yong-Soo;Lee, Yong-Ju;Kang, Ju-Il;Song, Jin-Won;Baek, Luck-Ju
    • The Journal of Korean Society of Virology
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    • v.28 no.4
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    • pp.337-345
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    • 1998
  • Hantaviruses are distributed in rodent population world-widely even in geographical areas where hemorrhagic fever with renal syndrome (HFRS) has not been reported. Various species of Family Muridae and Arvicolidae serve as the natural reservoirs of hantaviruses. Hantaan virus, Seoul virus, Puumala virus, Prospect HII virus, Sin Nombre virus and New York virus are members of genus Hantavirus and isolated from lungs of A. agrarius, R. norvegicus, C. glareolus, M. pennsylvanicus, P. maniculatus and P. leucopus respectively. This experiment was intended to find the distribution of hantavirus infection among wild rodents and isolate the hantavirus from lung tissue of seropositve Apodemus peninsulae, and compared the nucleotide and amino acid sequences with prototype of hantaan virus 76-118 strain. Hantaviral sequences were amplified from lung tissues of A. peninsulae by reverse-transcriptase polymerase chain reaction. Alignment and comparison of the 324 nucleotide of G2 region of M-genomic segment diverged 4.6% and 0% at the nucleotide and amino acid levels, and complete N protein-coding region of S-genomic segment diverged 3.7% and 1.4% nucleotide and amino acid levels, respectively. This is the report to spill-over on the hantaan virus from A. agrarius to A. peninsulae in Korea.

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Effect of Autoclave Curing on the Microstructure of Blended Cement Mixture Incorporating Ground Dune Sand and Ground Granulated Blast Furnace Slag

  • Alawad, Omer Abdalla;Alhozaimy, Abdulrahman;Jaafar, Mohd Saleh;Aziz, Farah Nora Abdul;Al-Negheimish, Abdulaziz
    • International Journal of Concrete Structures and Materials
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    • v.9 no.3
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    • pp.381-390
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    • 2015
  • Investigating the microstructure of hardened cement mixtures with the aid of advanced technology will help the concrete industry to develop appropriate binders for durable building materials. In this paper, morphological, mineralogical and thermogravimetric analyses of autoclave-cured mixtures incorporating ground dune sand and ground granulated blast furnace slag as partial cementing materials were investigated. The microstructure analyses of hydrated products were conducted using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), differential thermal analysis (DTA), thermo-graphic analysis (TGA) and X-ray diffraction (XRD). The SEM and EDX results demonstrated the formation of thin plate-like calcium silicate hydrate plates and a compacted microstructure. The DTA and TGA analyses revealed that the calcium hydroxide generated from the hydration binder materials was consumed during the secondary pozzolanic reaction. Residual crystalline silica was observed from the XRD analysis of all of the blended mixtures, indicating the presence of excess silica. A good correlation was observed between the compressive strength of the blended mixtures and the CaO/$SiO_2$ ratio of the binder materials.