• Title/Summary/Keyword: Fragment method

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The biochemical and molecular characteristics of Streptococcus equi subsp. zooepidemicus isolated from the genital tract of Thoroughbred mares in Korea

  • Choi, Seong-Kyoon;Kim, Seong-Guk;Cho, Gil-Jae
    • Korean Journal of Veterinary Service
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    • v.34 no.3
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    • pp.201-208
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    • 2011
  • Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a pathogen of a variety of infections in horse. We studied biochemical and molecular characteristics of S. zooepidemicus isolated from the genital tract of Thoroughbred mares in Korea. Seventy-nine isolates were identified as S. zooepidemicus by biochemical and PCR method from 374 horses. The biochemical characteristics of S. zooepidemicus isolates were positive reaction of lactose and sorbitol. However, S. zooepidemicus isoltes were negative reaction of inulin, mannitol, raffinose, trehalose, aesculin hydrolysis, growth in 6.5% NaCl and variable reaction of maltose. Epidemiological investigations of S. zooepidemicus isolates were performed by fragment analysis of SzP (S. zooepidemicus protective protein) gene, CNE (collagen binding protein) gene and ISR (16s rRNA intergenic spacer region) gene using ABI Prism $3,130{\times}1$ Genetic Analyzer System. All isolates were shown single amplification size of 906 bp in CNE gene, but SzP and ISR gene were shown variable patterns of fragment size. The characteristics of S. zooepidemicus investigated in this study will be very useful for the prevention of infection and the studies of epidemiologic characteristics of S. zooepidemicus, causing the severe economic losses due to reproductive failures.

Fragmentations and Proton Transfer Reactions of Product Ions Formed from Mono-, Di-, and Triethanolamines

  • Choi, Sung-Seen;So, Hun-Young
    • Bulletin of the Korean Chemical Society
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    • v.25 no.10
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    • pp.1538-1544
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    • 2004
  • Fragmentations and proton transfer reactions of mono-, di-, and triethanolamines were studied using FTMS. It was found that the most abundant fragment ion was $[M-CH_2OH]^+$. The $[M-CH_2OH-H_2O]^+$ was observed in the mass spectra of diethanolamine and triethanolamine. By increasing the ion trapping time in the ICR cell, the $[M+H]^+$ and $[M+H-H_2O]^+$ ions were notably increased for all the samples while the $[M+H-2H_2O]^+$ was observed in the mass spectra of diethanolamine and triethanolamine. The proton transfer reactions between the fragment ions and neutral molecules occurred predominantly by increasing the ion trapping time. The rate constants for the proton transfer reactions were calculated from experimental results. The proton transfer reaction of $CHO^+$ was the fastest one, which is consistent with the heats of reaction. The rate constants for proton transfer reactions of triethanolamine were much slower than those of ethanolamine and diethanolamine because of the steric hindered structure of triethanolamine. The plausible structures of observed ions and heats of reaction for proton transfer were calculated with AM1 semiempirical method.

Hinge Technique in One Stage Reduction Cranioplasty for Macrocephaly with Hydrocephalus - A Case Report - (수두증을 동반한 거대두개증 환아에서 격자술을 이용한 One Stage Reduction Cranioplasty - 증례보고 -)

  • Weon, Keun Soo;Yoon, Soo Han;Shin, Yong Sam;Cho, Ki Hong;Cho, Kyung Gi
    • Journal of Korean Neurosurgical Society
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    • v.30 no.sup1
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    • pp.159-164
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    • 2001
  • Reduction cranioplasty is one of the treatment modality among many treatment options for macrocephaly with hydrocephalus. The most previous techniques of reduction cranioplasty have some disadvantages such as difficult airway maintenance, pressure sore due to modified prone position, severe venous infarct due to obstruction of venous blood flow and large amount of bleeding from the dissection of superior sagittal sinus to obtain bone fragment needed. A 28-month-old girl had extreme macrocephaly. She couldn't control head rotation and keep sitting position. The operation was performed at supine position with adequate exposure of entire calvarium and the hinge was made in occipital bone fragment that covered posterior part of superior sagittal sinus. Bleeding volume and the other complications were decreased and acceptable reduction was achieved with this method.

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Theoretical Studies on the Gas-Phase Pyrolysis of Esters The effect of ${\alpha}$- and ${\beta}$-methylation of Ethyl Formates

  • Ikchoon Lee;Ok Ja Cha;Bon-Su Lee
    • Bulletin of the Korean Chemical Society
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    • v.11 no.1
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    • pp.49-54
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    • 1990
  • The gas-phase thermolysis reactions of ${\alpha}$- and ${\beta}$-methylated ethyl formates, Y = $CH-X-CHR_1CH_2R_2$ where X = Y = O or S and $R_1\;=\;R_2$ = H or $CH_3$, are investigated theoretically using the AM1 method. The experimental reactivity order is reproduced correctly by AM1 in all cases. The thermolysis proceeds through a six-membered cyclic transition state conforming to a retro-ene reaction, which can be conveniently interpreted using the frontier orbital theory of three-species interactions. The methyl group substituted at $C_{\alpha}\;or\;C_{\beta}$ is shown to elevate the ${\pi}$-HOMO of the donor fragment (Y = C) and depress the ${\sigma}^{\ast}$-LUMO of the acceptor fragment ($C_{\beta}$-H), increasing the nucleophilicity of Y toward ${\beta}$-hydrogen which in turn increases the reactivity. The two bond breaking processes of the $C_{\alpha}$-X and $C_{\beta}$-H bonds are concerted but not synchronous so that the reaction takes place in two stages as Taylor suggested. The initial cleavage of $C_{\alpha}$-X is of little importance but the subsequent scission of $C_{\beta}$-H occurs in a rate determining stage.

Characterization of the Fragmentation Pattern of Peptide from Tandem Mass Spectra

  • Ramachandran, Sangeetha;Thomas, Tessamma
    • Mass Spectrometry Letters
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    • v.10 no.2
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    • pp.50-55
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    • 2019
  • The fragmentation statistics of ion trap CID (Collision-Induced Dissociation) spectra using 87,661 tandem mass spectra of doubly charged tryptic peptides are analyzed here. In contrast to the usual method of using intensity information, the frequency of occurrence of fragment ions, with respect to the position of the cleavage site and the residues at these sites is studied in this paper. The analysis shows that the frequency of occurrence of fragment ion peaks is more towards the middle of the peptide than its ends. It was noted that amino acid with an aromatic and basic side chain at N- & C- terminal end of the peptide stimulates more peaks at the lower end of the spectrum. The residue pair effect was shown when the amide bond occurs between acidic and basic residues. The fragmentation at these sites (D/E-H/R/K) stimulates the generation of the y-ion peak. Also, the cleavage site H-H/R/K stimulates the generation of b-ions. K-P environment in the peptide sequence has more tendency to generate y-ions than b-ions. Statistical analysis helps in the visualization of the CID fragmentation pattern. Cleavage pattern along the length of the peptide and the residue pair effects, enhance the knowledge of fragmentation behavior, which is useful for the better interpretation of tandem mass spectra.

Purification and Gene Analysis of Peptidyl Prolyl cia-trans Isomerase from Bacillus stearothermophilus (Bacillus stearothermophilus Peptidyl Prolyl cis-trans Isomerase의 정제 및 유전자 분석)

  • 김동주
    • The Korean Journal of Food And Nutrition
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    • v.15 no.2
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    • pp.104-111
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    • 2002
  • The peptidyl prolyl sis-trans isomerase (PPIase, EC 5.2.1.8) from bacillus stearothermophilus was extracted from the cells treated with by lysozyme. PPIase was purified from the cell extracts by heat treatment, ammonium sulfate precipitation, ion exchange chromatography and finally gel filtration, sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The molecular weight of the purified PPIase was estimated as 18kDa by SDS-PAGE. The 39 amino acid residues from the N-terminus were determined by the protein sequencer. The enzyme showed the optimum pH at 8.0 and was stable at the range of pH 7.0∼8.0. The enzyme was considerably stable after heat treatment at 60$\^{C}$ for 30minutes, and the enzyme was quite stable up to 65$\^{C}$. The presence of the PPIase in the refolding solution accelerated the isomerization rate of the assay peptide. PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-l primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPI-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus.

INTRA-ALVEOLAR TRANSPLANTATION OF COMPLETELY CROWN-ROOT FRACTURED TOOTH WITH DEMINERALIZED FREEZED DRIED BONE GRAFT (치은연 하방으로 파절된 치아의 탈회냉동건조골을 이용한 Intra-alveolar transplantation)

  • Lim, Hyoung-Soo;Kim, Dong-Phil;Lee, Chang-Seop;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.2
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    • pp.344-350
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    • 2000
  • Incidence of crown-root fracture due to traumatic injury, have been reported 3% in the permanent dentiton, 2% in the deciduous dentition. There are two treatment methods for crown-root fractured teeth with pulp exposure, when the fracture line was located under the alveolar crest. One way is the extrusion by orthodontic force the other way is intra-alveolar transplantation which occlusally repositioning of apical fragment in the alveolar socket. Since intra-alveolar transplantation has introduced in 1970s, it was practiced as alternative to orthodontic extrusion. As the result, this method may thoughted that had a good prognosis. As a result of trauma, completely crown-root fracture was occured in the maxillary right central incisor in this case. We couldn't reposition the deepest fracture line above the alveolar crest by the conventional surgical extrusion, because apical fragment was too short. Thus, after extraction of apical fragment, we repositioned it to the socket following demineralized freezed dried bone graft, which possible to support the apical fragment. At the 15-month recall examination, the root still showed normal mobility and there was not observed any in flammatory or replacement root resorption in the periapical radiograph.

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Gene Cloning, Nucleotide Sequence and Efficent Expression of Peptidyl proryl cis-trans Isomerase from Bacillus stearothermophilus (Bacillus stearothermophilus의 Peptidyl Prolyl cis-trans Isomerase 유전자 분리 염기배열 및 발현)

  • 김동주
    • The Korean Journal of Food And Nutrition
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    • v.9 no.4
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    • pp.452-458
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    • 1996
  • A PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-1 primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI1-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPi-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus. The translated protein sequence of PPIase B. stearothermophilus was compared with sequence from periplasmic PPIase from Escherichina coil ; homogies of 16 and 58%, respectively, were found. The clond PPIase gene was over-expressed in E. coil cell using pUC19 as an expression vector. The enzyme was partially purified by heat treatment and colum chromatochraphy on DEAE-Sepharose CL-6B. The molecular weight of the enzyme was dermined to be about 18.0 kDal by SDS-PAGE.

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Application of Methodology for Microbial Community Analysis to Gas-Phase Biofilters (폐가스 처리용 바이오필터에 미생물 군집 분석 기법의 적용)

  • Lee, Eun-Hee;Park, Hyunjung;Jo, Yun-Seong;Ryu, Hee Wook;Cho, Kyung-Suk
    • Korean Chemical Engineering Research
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    • v.48 no.2
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    • pp.147-156
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    • 2010
  • There are four key factors for gas-phase biofilters; biocatalysts(microorganisms), packing materials, design/operating techniques, and diagnosis/management techniques. Biofilter performance is significantly affected by microbial community structures as well as loading conditions. The microbial studies on biofilters are mostly performed on basis of culture-dependent methods. Recently, advanced methods have been proposed to characterize the microbial community structure in environmental samples. In this study, the physiological, biochemical and molecular methods for profiling microbial communities are reviewed, and their applicability to biofilters is discussed. Community-level physiological profile is based on the utilization capability of carbon substrate by heterotrophic community in environmental samples. Phospholipid fatty acid analysis method is based on the variability of fatty acids present in cell membranes of different microorganisms. Molecular methods using DNA directly extracted from environmental samples can be divided into "partial community DNA analysis" and "whole community DNA analysis" approaches. The former approaches consist in the analysis of PCR-amplified sequence, the genes of ribosomal operon are the most commonly used sequences. These methods include PCR fragment cloning and genetic fingerprinting such as denaturing gradient gel electrophoresis, terminal-restriction fragment length polymorphism, ribosomal intergenic spacer analysis, and random amplified polymorphic DNA. The whole community DNA analysis methods are total genomic cross-DNA hybridization, thermal denaturation and reassociation of whole extracted DNA and extracted whole DNA fractionation using density gradient.

Synthesis of 5-Fluorouracil by Ring Transformation of s-Triazine (s-Triazine의 Ring Transformation에 의한 5-FU의 합성)

  • 정원근;정진현
    • YAKHAK HOEJI
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    • v.26 no.1
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    • pp.25-27
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    • 1982
  • We had reported that s-triazine can readily be converted into the corresponding 5-substituted pyrimide. In order to develop new synthetic method of 5-fluorouracil, we tried to replace eliminating fragment, 1, 3-dimethylurea, by fluoroacetamide, which was expected to undergo nucleophilic attack by proton extraction of both .alpha.-hydrogen and aminohydrogen by lithium diisopropylamide (LDA). We found that 5-fluorouracil could be transformed from s-triazine under strong base condition like LDA as well as other 5-substituted pyrimidines.

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