• Title/Summary/Keyword: Fragment method

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CLINICO-STATISTICAL ANALYSIS OF POSSIBLE FACTORS LEADING TO PROBLEMS IN THE SURGICAL TREATMENT OF UNILATERAL MANDIBLAR CONDYLE FRACTURES (편측 하악 과두골절의 관혈적 치료에 있어서 예후에 영향을 줄 수 있는 인자들에 관한 임상 통계학적 연구)

  • Sung, Hun-Mo;Lee, Dong-Keun;Min, Seung-Ki;Oh, Seung-Hwan;Jang, Kwan-Sik
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.23 no.1
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    • pp.31-39
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    • 2001
  • The purpose of this study is to reveal the factors leading to the problem of unilateral condylar fractures and suggest a treatment guideline of treatment for good prognosis in surgical treatment. The factors can be age, sex, fracture site, degree of displacement, posterior occlusion loss, post-operative alteration of condylar head position, post-operative condylar head resorption, and maxillomandibular fixation period. One hundred and eleven patients with unilateral condylar fractures, who were treated by surgical method from 1990 Feb. to 2000 Feb., were studied. Minimum follow-up period was 6 months. The results were as follows ; 1. In the age group of $41{\sim}60$, females had significantly higher complication rate than males, therefore we must be careful about treatment of female in this age group 2. In level I fractures of the mandibular condyle, because there were abundant complications when the patients were treated with fragment removal, conservative treatment is recommended over the surgical approach. 3. There were no differences in the complication rate, in the level II, III fractures. but were severe complications in the cases of patients treated by Dr.Nam's method or fragment removal. Therefore, open reduction and internal fixation is recommended over Dr.Nam's method or fragment removal. 4. In level IV fractures, open reduction and internal fixation is recommended 5. Although there was a higher complication rate depending on the degree of deviation, there was no correlation between the degree of deviation and development of complications in each level of fracture 6. Because the complication rate was higher in cases of condylar resorption, vertical dimension loss, and alteration of condylar head position, we must make an effort to prevent such complications during treatment

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Development of RT-PCR Kit for Diagnosis of Pathogenic Agent of Ginseng Root Rot in the Ginseng Field (인삼포장에서 뿌리섞음병원균의 진단을 위한 RT-PCR KIT의 개발)

  • 도은수
    • Korean Journal of Plant Resources
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    • v.16 no.1
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    • pp.40-48
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    • 2003
  • Cylindrocarpon destructans is the major pathogen inducing the root rot disease in ginseng. Up to now, there is no reliable and convenient method to analyze the spore density or population of this pathogen in ginseng-growing soil or any contaminated farmlands. Therefore, it will be very valuable to develop a new and reliable method in detecting the spore of this pathogen. In this study, a molecular biological technique using two step nested PCR method, was developed. Two universal ITS primers, ITS5F and ITS4R were used in the first round of PCR to amplify a fragment of ITS region from the genomic DNA of C. destructans. The specific prmers Nest 1 and Nest 2 were designed and used in the second round of PCR to amplify a inner fragment from the first round PCR product of C. destructans. C. destructans spore, only soil samples from the diseased ginseng farm produced the positive bands, suggesting its usefulness in detecting the C. destructans spores in soil samples. Thus it is recommended to first extract the whole genomic DNA from soil samples and use it for the PCR reaction, thereby eliminating the inhibitory activity of soil components.

Theoretical Study on the Reaction Mechanism of Azacyclopropenylidene with Epoxypropane: An Insertion Process

  • Tan, Xiaojun;Wang, Weihua;Li, Ping
    • Bulletin of the Korean Chemical Society
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    • v.35 no.9
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    • pp.2717-2722
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    • 2014
  • The reaction mechanism between azacyclopropenylidene and epoxypropane has been systematically investigated employing the second-order M${\o}$ller-Plesset perturbation theory (MP2) method to better understand the reactivity of azacyclopropenylidene with four-membered ring compound epoxypropane. Geometry optimization, vibrational analysis, and energy property for the involved stationary points on the potential energy surface have been calculated. It was found that for the first step of this reaction, azacyclopropenylidene can insert into epoxypropane at its C-O or C-C bond to form spiro intermediate IM. It is easier for the azacyclopropenylidene to insert into the C-O bond than the C-C bond. Through the ring-opened step at the C-C bond of azacyclopropenylidene fragment, IM can transfer to product P1, which is named as pathway (1). On the other hand, through the H-transferred step and subsequent ring-opened step at the C-N bond of azacyclopropenylidene fragment, IM can convert to product P2, which is named as pathway (2). From the thermodynamics viewpoint, the P2 characterized by an allene is the dominating product. From the kinetic viewpoint, the pathway (1) of formation to P1 is primary.

The Relative Identification of C. officinale and L. chuanxiong by PCR-Mediated Fingerprinting (천궁류(川芎類) 한약재의 유전자 감식 연구)

  • Choi, Ho-Young;Kim, Dong-Wook;Kim, Dong-Eun;Suh, Young-Bae;Ham, In-Hye
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.151-161
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    • 2005
  • Objectives : Our research purpose is to establish the standard identification analysis on C. officinale and L. chuanxiong in Korea and China by PCR-mediated fingerprinting. Methods : The Restriction Fragment Length Polymorphism (RFLP) and Randomly Amplified Polymorphic DNA (RAPD) method was used on Internal Transcribed Spacer (ITS) regions and rbcL regions to compare and discriminate genes extracted from crude drugs as C. officinale and L. chuanxiong in Korea and China. Results : L. chuanxiong Korea and China have very similar polymorphism, whereas L. chuanxiong in Korea and C. officinale have very different polymorphism in RFLP. And restriction enzymes AluI and SacI forms the specific fragment band only in C. officinale, they can be used as RFLP marker on ITS regions to discriminate among the species. Conclusions : The results could be applied in discriminating crude drugs among C. officinale and L. chuanxiong in Korea and China. Also they could be used in controlling drug quality, preserving medicinal plants, and improving plant description.

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Identification of Mycobacteria by Comparative Sequence Apalysis and PCR-Restriction Fragment Length Polymorphism Analysis (염기서열과 PCR-Restriction Fragment Length Polymorphism 분석에 의한 Mycobacteria 동정)

  • Kook, Yoon-Hoh
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.6
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    • pp.561-571
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    • 1999
  • Diagnosis of mycobacterial infection is dependent upon the isolation and identification of causative agents. The procedures involved are time consuming and technically demanding. To improve the laborious identification process mycobacterial systematics supported by gene analysis is feasible, being particularly useful for slowly growing or uncultivable mycobacteria. To complement genetic analysis for the differentiation and identification of mycobacterial species, an alternative marker gene, rpoB encoding the ${\beta}$ subunit of RNA polymerase, was investigated. rpoB DNAs (342 bp) were amplified from 52 reference strains of mycobacteria including Mycobacterium tuberculosis H37Rv (ATCC 27294) and clinical isolates by the PCR. The nucleotide sequences were directly determined (306 bp) and aligned using the multiple alignment algorithm in the MegAlign package (DNASTAR) and MEGA program. A phylogenetic tree was constructed with a neighborhood joining method. Comparative sequence analysis of rpoB DNA provided the basis for species differentiation. By being grouped into species-specific clusters with low sequence divergence among strains belonging to same species, all the clinical isolates could be easily identified. Furthermore RFLP analysis enabled rapid identification of clinical isolates.

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Development Strategy for functional rice improved with human lactoferrin and enhancement of nutrient compounds (인체 모유 단백질 및 영양 성분 강화 고부가가치 기능성 쌀 생산 벼 품종 개발 전략)

  • Rhim Seong-Lyul;Lee Jin-Hyoung;Lee Hyo-Yeon;Suh Suk-Cheol
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2002.05a
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    • pp.48-50
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    • 2002
  • A strategy for development of a functional rice in proved with human lactoferrin and enhancement of nutrient compounds was planned. For the purposes we have cloned and characterized a human lactoferrin cDNA from human mammary gland cDNA library A endosperm storage vacuole targeting sequence and the cDNA fragment was linked to endosperm specific glutelin promoter. The fusion gene fragment was inserted into a binary vector containing MAR gene. In addition a new ${\beta}$-galactosidase gene from Bifidobacterium of human was used as a reporter gene in the vector system, Rice plants showing a high concentration of amino acids in the endosperm cells were developed by using a biochemical mutation and bred for the transformation with the binary vector system Finally we have established a transformation method for the rice endosperm cells.

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Paratope Mapping of Anti-Ex-A IgG as Studied by NMR (NMR에 의한 anti-Ex-A IgG의 항원결합부위 해석)

  • Kim, Ha-Hyeong;Lee, Gwang-Pyo
    • YAKHAK HOEJI
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    • v.40 no.4
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    • pp.422-427
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    • 1996
  • The anti-Ex-A IgG was specifically labeled with stable isotopes, DL-His-2,4-$d_2$, L-Phe-$d_5$, L-Trp-$d_5$, L-Tyr-2,6-$d_2$ and L-[1-$^{13}C$]Trp, by growing hybridoma cell in serum-free medium. By use of NMR spectroscopy with selectively labeled Fab fragment, we applied a paratope mapping on antigen-antibody complex. Assignments of the observed carbonyl carbon resonances have been determined by using $^{13}C$-$^{15}N$ double labeling method in order to assign the Trp resonances. Photo CIDNP was also applied to investigate the antigen-binding site(s) on the surface residues of antibody. We found that Trp 36, which is located at the $V_H$ domain, is an important residue to bind to Ex-A, however, two Tyr on the surface of anti-Ex-A IgG plays no crucial role to bind to antigen. On the basis of these results, we demonstrate that stable isotope-aided NMR strategy can be extended to molecular structural analyses of the complex of an Fab fragment and a protein antigen.

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A Rapid and Simple Method for Construction and Expression of a Synthetic Human Growth Hormone Gene in Escherichia coli

  • Roytrakul, Sittiruk;Eurwilaichitr, Lily;Suprasongsin, Chittiwat;Panyim, Sakol
    • BMB Reports
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    • v.34 no.6
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    • pp.502-508
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    • 2001
  • A cDNA, encoding the human growth hormone (hGH), was synthesized based on the known 191 amino acid sequence. Its codon usage was optimized for a high level expression in Escherichia coli. Unique restriction sites were incorporated throughout the gene to facilitate mutagenesis in further studies. To minimize an initiation translation problem, a 624-bp cassette that contained a ribosome binding site and a start codon were fused to the hGH-coding sequence that was flanked between the EcoRI and HindIII sites. The whole fragment was synthesized by an overlapped extension of eight long synthetic oligonucleotides. The four-short duplexes of DNA, which were first formed by annealing and filling-in with a Klenow fragment, were assembled to form a complete hGH gene. The hGH was cloned and expressed successfully using a pET17b plasmid that contained the T7 promoter. Recombinant hGH yielded as much as 20% of the total cellular proteins. However, the majority of the protein was in the form of insoluble inclusion bodies. N-terminal amino acid sequencing also showed that the hGH produced in E. coli contained formyl-methionine. This study provides a useful model for synthesis of the gene of interest and production of recombinant proteins in E. coli.

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Synthesis of Cyclic Peticdes Related to GBⅠ-Ⅱand Study of Their Inhibitory Activity for Proteinases (GBⅠ-Ⅱ 와 관련된 Cyclic Peptide 들의 합성과 단백질 분해 효소에 대한 저해활성 연구)

  • Gang, Sin Won;Heo, Nam Won
    • Journal of the Korean Chemical Society
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    • v.34 no.3
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    • pp.288-296
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    • 1990
  • Antielastic fragment in GBⅠ-Ⅱ differ on $P_1$ site with that of antitryptic fragment in LBI. To obtain further understanding of the role of amino acid residue near the reactive site, specificity of $P_1$ site and loop size, Tyr substituted cyclic nonapeptide and cyclic pentapeptide were synthesized and the inhibition constants for some proteinases were calculated by Dixon method. Cyclic nonapeptide showed no inhibition for chymotrypsin but appeared low inhibitory activity for trypsin and elastase and that of cyclic pentapeptide possessed inhibitory activity for chymotrypsin.

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Identification of Meat Species Using PCR-RFLP Marker of Cytochrome b Gene

  • Shin, Sung-Chul;Chung, Ku-Young;Chung, Eui-Ryong
    • Food Science of Animal Resources
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    • v.26 no.3
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    • pp.375-379
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    • 2006
  • Food labeling regulations require that the meat species in various meat products are accurately declared to the consumer. Substitution or adulteration of costly meat with a cheaper one is one of the most common problems in the meat industry. In this study, PCR-restriction fragment length polymorphism(RFLP) method of the mitochondrial cytochrome b(mt cyt b) gene has been applied for identification of the origin of six mammalian meat species(beef, port horse, goat, mutton and deer) and three poultry meat species(chicken, turkey and duck) as raw materials for meat products. PCR was used to amplify a variable region of mt cyt b gene. Meat species differentiation was determined by digestion of the amplified products with a 359 bp fragment using HaeIII and HinfI restriction enzymes, which generated species-specific RFLP patterns. This PCR-RFLP DNA marker of mt cyt b gene could be very useful for the accurate and reliable identification and discrimination of animal meat species in routine analysis.