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Cytogenetic and Molecular Genetic Studies on Duchenne Muscular Dystrophy (Duchenne Muscular Dystrophy에 관한 세포유전학 및 분자유전학적 연구)

  • Hong, Hea-Sook
    • Journal of Korean Biological Nursing Science
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    • v.7 no.1
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    • pp.29-46
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    • 2005
  • Purpose ; 본 연구는 X-염색체와 관련된 장애 중에서 가장 흔하고 심한 Duchenne Muscular Dystrophy(DMD)의 세포유전학 및 분자유전학적 특성을 설명하기 위해서 DMD에 영향을 받고 있는 두 가계의 13명을 대상으로 가계도 분석과 염색체 분석 및 DNA 분석을 하였다. Method ; DNA분석은 DNA probe을 이용한 Southern blotting method로써 RFLPs와 DMD유전자 부위의 exon소실 유무를 조사하여 아래와 같은 결과를 얻었다. Conclusion ; A 염색체 분석 : 말초혈액과 양수를 표본으로 High-Resolution GTG염색에서 A가계와 B가계의 염색체 분석에서 12명의 염색체는 정상 X-염색체였으나 B가계의 I-2(DMD여성)에서 46, x,-x,+t(2:x)(q 21.1 : p21.2)로 나타난다. B. DNA분석3 : 1) RFLPs의 분석 J66,XJ-1.1,754-11로써 B가계의 RELPs(Restriction Fragment Length Polymorphisms)에서 J66/Pst I은 1.7hb(E), 1.6kb(e)을 보여 주었고 XJ-1.1/Taq I은 3.6kb(F), 3.0kb(f), 754-11/EoR I은 4.2kb(G), 2.0kb(g)의 대립인자를 나타내었다. 이상의 결과를 바탕으로 영향을 받고 있는 남자 (II-2)의 haplotype는 보인자인 어머니의 한쪽 인자를 받았으며 어머니와 딸은 보인자이고 임산부의 태아는 남아였고 태아의 인자들은 그의 할아버지로부터 물려받아 DMD에 영향을 받지 않은 것으로 진단되었다. 2) DMD 유전자의 exon 소실에 대한 분석 cDNA probe 8과 cDNA probe 2b-3으로써 소실에 대한 진단은 영향을 받은 남자(II-2)는 cDNA probe 8에서 12, 7.3, 6.6, 4.2kb에 소실이 있고 cDNA 2b-3은 1.7kb에 소실에 나타났다.

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Expression of Cyclin D3 Transcripts in the Postmeiotic Male Germ Cells of the Mouse

  • Sun, Woong-Sun;Geum, Dong-Ho;Choi, Wan-Sung;Kim Kwon, Yun-Hee;Rhee, Kun-Soo;Kim, Kyung-Jin
    • Animal cells and systems
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    • v.2 no.4
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    • pp.495-500
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    • 1998
  • D-type G1 cyclins are known to be crucial for the progression of mitotic cell cycle in mammals. Although many studies have been performed to elucidate the roles of D-type cyclins, it is largely unknown whether D-type cyclins are directly involved in the regulation of meiotic germ cell development. In the present study, we examined the expression patterns of D-type cyclins (cyclin D1 and D3) during male germ cell development by northern blot and in situ Hybridization analyses. In the adult testes, we detected a 4.2 kb cyclin D1 mRNA and two different sizes (2.3 kb and 1.8 kb) of cyclinD3 mRNAs. The short form of the cyclin D3 transcript was testis-specific. Along with the testicular development, expression of cyclin D3 mRNA was increased whereas cyclin D1 mRNA was gradually decreased. in situ hybridization study also revealed that the expression of cyclin D3 was restricted to the postmeiotic germ cells. Furthermore, the 2.3 kb transcript was highly expressed in the round spermatids and decreased in the elongated spermatids/residual bodies, while the 1.8 kb transcript was expressed in elongated spermatids/residual bodies more abundantly. Sucrose-gradient separation of polysomal RNA fractions demonstrated that some portions of the 2.3 kb transcript are translationally active, while the 1.8 kb transcript is likely to be inactive. Taken together, the present data suggest a functional importance of cyclin D3 expression in the differentiated postmeiotic male germ cells.

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Antioxidative Activity and Cytotoxicity on Human KB cell of Extracts from Rhododendron mucronulatum Turcz. Flower (진달래꽃 추출물의 항산화 효과 및 인체 KB cell에 대한 세포독성)

  • Park Seung-Woo;Kim Sang-Gyo;Kim Mee-Jeoung
    • Food Science and Preservation
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    • v.13 no.4
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    • pp.501-505
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    • 2006
  • This study was carried out to investigate the antioxidative activity and cytotoxicity on human KB cell of extract from Rhododendron mucronulatum flower, and also to determine the contents of total polyphenol and flavonoid. The methanol extract of Rhododendron mucronulatum flower was fractionated with various solvents such as hexane, chloroform, ethyl acetate, butanol and water. The DPPH radical scavenging activities of ethyl acetate fraction had stronger activity than other fractions. So, antioxidative substances of ethyl acetate fraction were crude purified by silica gel column chromatography. The DPPH radical scavenging activities of crude purified fraction 2 and 3 were more than 90% at 40 ppm. In the presence of $100{\mu}g/mL$, growth inhibition on human KB cell by WST-1 assay showed 81.2% in chloroform fraction and 74.6% in hexane fraction. The contents of total polyphenol and flavonoid of ethyl acetate traction were 32.70% and 20.30%, respectively, The antioxidative activity showed correlation with total polyphenol and flavonoid contents.

Isolation and Evaluation of Protective Effect against Fusarium Wilt of Sesame Plants of Antibiotic Substance from Bacillus polymyxa KB-8

  • Hyun, Jae-Wook;Kim, Young-Hoon;Lee, Yong-Se;Park, Won-Mok
    • The Plant Pathology Journal
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    • v.15 no.3
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    • pp.152-157
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    • 1999
  • An antibiotic compound was isolated from the culture of an antagonist against Fusarium oxysporum f. sp. sesami, Bacillus polymyxa strain KB-8, and tested for the control of Fusarium wilt of sesame in greenhouse conditions. Optimum conditions for culturing the antagonist to obtain the maximum antibiotic activity were determined using different culture media, initial medium acidity, and incubation periods for which yeast -malt extract agar with the initial acidity of pH 5 and over 13 days culture were best. Antibiotic substances extracted by methanol had 2 main fractions, KB-8A and KB-8B, in thin layer chromatography (OLC) with Rf values of 0.35 and 0.67 in a solvent system of chloroform : methanol = 7 : 3. The fraction KB-8A wa purified further by XAD-2, silica gel and Sephadex LH-20 column chromatography, and crystalization. Its minimum inhibitory concentrations (MICs) were $12.8\mu\textrm{g}$/ml for F. oxysporum and Alternaria mali, $6.4\mu\textrm{g}$/ml for Colletotrichum gloeosporioides and Rhizoctonia solani, and $3.2\mu\textrm{g}$/ml for Phytophthora capsici. Soil drenching of antibiotic KB-8A in the concentrations of $13.0\mu\textrm{g}$/ml and $26.0\mu\textrm{g}$/ml effectively inhibited the Fusarium wilt of sesame in a greenhouse test, which appeared to be comparable to the fungicide benlate of $6.5\mu\textrm{g}$ a. i./ml.

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Molecular Cloning and Expression of Bacillus stearothermophilus $\beta$-D-Xylosidase Gene in E. coli (Bacillus stearothermophilus $\beta$-D-Xylosidase 유전자의 크로닝 및 Escherichia coli에서의 발현)

  • 오세욱;박성수;최용진;박영인
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.136-142
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    • 1992
  • Bacillus stearothemophilus isolated from soil was identified to express multiple extracellular xylanases. Two HindIII restriction fragments of 5.4 and 6.4 kb from B. stearothermophilus genomic DNA were cloned into pBR322 to obtain recombinant plasmids pMG0l and pMG02, respectively, which enabled E. coli HBlOl cells to produce $\beta$-D-xylosidase activity. By subcloning into pUC18 and Southern blotting, the loci of the $\beta$-D-xyiosidase genes were elucidated to be on non-homologous DNA fragments of 2.2 kb from pMGOl(pMG1) and 1.0 kb from pMG02(pMG2), respectively. The two enzymes produced in E. coli cleaved xylobiose, xylotriose, xylotetrose and xylotetrose to produce xylose as a major end product. The gene on pMG1, distinct from that on pMG2 was observed to encode a bifunctional protein that displayed both P-D-xylosidase (EC.3.2.1.37) and a-L-arabinofuranosidase activities (EC.3.2.1.55).

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Numerical Modeling of Tide and Tidal Current in the Kangjin Bay, South Sea, Korea

  • Ro, Young-Jae;Jun, Woong-Sik;Jung, Kwang-Young;Eom, Hyun-Min
    • Ocean Science Journal
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    • v.42 no.3
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    • pp.153-163
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    • 2007
  • This study is based on a series of numerical modeling experiments to understand the tidal circulation in the Kangjin Bay (KB). The tidal circulation in the KB is mostly controlled by the inflow from two channels, Noryang and Daebang which introduce the open ocean water into the northern part of the KB with relatively strong tidal current, while in the southern part of the KB, shallowest region of the entire study area, weak tidal current prevails. The model prediction of the sea level agrees with observed records at skill scores exceeding 90 % in terms of the four major tidal constituents (M2, S2, K1, O1). However, the skill scores for the tidal current show relatively lower values of 87, 99, 59, 23 for the semi-major axes of the constituents, respectively. The tidal ellipse parameters in the KB are such that the semi-major axes of the ellipse for M2 range from 1.7 to 38.5 cm/s and those for S2 range from 0.5 to 14.4 cm/s. The orientations of the major-axes show parallel with the local isobath. The eccentricity values at various grid points of ellipses for M2 and S2 are very low with 0.2 and 0.06 on the average, respectively illustrating that the tidal current in the KB is strongly rectilinear. The magnitude of the tidal residual current speed in the KB is on the order of a few cm/s and its distribution pattern is very complex. One of the most prominent features is found to be the counter-clockwise eddy recirculation cell at the mouth of the Daebang Channel.

A study for a construction of Theileria sergenti DNA probe (Theileria sergenti DAN probe를 만들기 위한 기초 연구)

  • Kim, Myeong-chul;Lee, Joo-mook;Kwon, Oh-deog;Chae, Joon-seok;Kim, Heung-seob
    • Korean Journal of Veterinary Research
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    • v.33 no.3
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    • pp.479-486
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    • 1993
  • This study was attempted co develope a method for detection of Theileria sergenti infection on the basis of hybridization of parasite DNA with a probe. For construction of a T sergenti genomic library, T sergenti DNA was digested completely with Bam-HI and the fragments were ligated into the Bam-HI site of pUC-19 before transformation of Escherichia colistrain JM83. To detect clones containing the parasite's DNA sequences, a genomic DNA library of T sergenti constructed in pUC-19 was screened by cracking and Southern hybridization. Seven colonies were chosen from 29 colonies which were screened by transformation of Escherichia coli strain JM83. Seven transformants were comfirmed from seven colonies by cracking. The sizes of transformants were about 5Kb, 5.7Kb, 4.3Kb, 7.75Kb, 7.85Kb, 5.8Kb, 3.8Kb, respectively. DNA inserts, T sergenti DNA, and bovine DNA were hybridized with radio-labelled T sergenti DNA. Two($pT_1$, $pT_1$) of the seven inserts and T sergenti DNA reacted strongly but another 5 inserts and bovine DNA showed weak reation. All of the DNA inserts were not reaction, but T sergenti DNA were very weakly and bovine DNA were strongly reacted to hybridization with radio-labelled bovine DNA. Therefore, we obtained total 7 T sergenti DNA fragments in this study.

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Structure of Spodoptera exigua Nucleopolyhedrovirus p10 Gene (파밤나방 핵다각체병 바이러스의 p10 유전자 구조)

  • 최재영;우수동;홍혜경;이해광;제연호;강석권
    • Korean journal of applied entomology
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    • v.38 no.2
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    • pp.145-149
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    • 1999
  • To develop the baculovirus expression vector system (BEVS) adopting p10 gene promoter of Spodoptera exigua nucleopolyhedrovirus (SeNPV), we characterized the p10 gene of SeNPV. The nucleotide sequence of 545 bases including the coding region of p10 gene was determined. Compared with the previously reported SeNPV p10 gene (Zuidema et al., 1993), 4 bases were different in the 5' and 3' flanking region but no difference was found in the coding region. The p10 gene was located within Xho I 1.5 Kb, Sph 1 2.4 Kb and Cla I 4.0 Kb fragments by Southern hybridization analysis. Also, the Sph I 2.4 Kb and the Cla I 4.0 Kb fragments were cloned and their restriction enzyme maps were determined.

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A STUDY ON THE RADIOSENSITIVITY AND CHEMOSENSITIVITY OF KB CELL LINE IN VITRO (시험관내 KB세포주의 방사선 및 항암제감수성에 관한 연구)

  • Hong Sung-Woo;Choi Eun-Suk;Koh Kwang-Joon
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.28 no.1
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    • pp.271-283
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    • 1998
  • The purpose of this study was to aid in the prediction of tumor cell tolerance to radiotherapy and/or chemotherapy. For this study, cell surviving curves were obtained for human squamous cell carcinoma KB cell line after radiation exposure and/or administration of antitumor drugs. 2, 4, 6, 8, 10Gy were irradiated at a dose rate of 210cGy/min using /sup 60/Co Irradiator ALDORADO 8. After irradiation, KB cell lines (3×104cells/ml) were exposed to 2㎍/ml of bleomycin or cisplatin for 1 hour. The viable cells were determined by MTT assay for each radiation dose and/or each drug at the 4th day. And they were compared to control values. The obtained results were as follows : 1. The slope of the surviving curve after irradiation of 2, 4, 6, 8, 10Gy on KB cell line was relatively steep. 2. There was no significant difference between the cytotoxicity of bleomycin compared to control group. But, there was significant difference between the cytotoxicity of cisplatin compared to control group. And the cytotoxicity of cisplatin was greater than that of bleomycin on KB cell line. 3. There were significant differences of surviving fractions after irradiation of 2Gy and 10Gy with 2㎍/ml of bleomycin compared with the groups of irradiation only on KB cell line. 4. There were significant differences of surviving fractions after irradiation of 2, 4, 6, 8, 10Gy with 2㎍/ml of cisplatin compared with the groups of irradiation only on KB cell line. 5. There was significant difference of surviving fraction between groups after irradiation of 10Gy with 2㎍/ml of bleomycin and cisplatin.

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An Area Efficient Low Power Data Cache for Multimedia Embedded Systems (멀티미디어 내장형 시스템을 위한 저전력 데이터 캐쉬 설계)

  • Kim Cheong-Ghil;Kim Shin-Dug
    • The KIPS Transactions:PartA
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    • v.13A no.2 s.99
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    • pp.101-110
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    • 2006
  • One of the most effective ways to improve cache performance is to exploit both temporal and spatial locality given by any program executional characteristics. This paper proposes a data cache with small space for low power but high performance on multimedia applications. The basic architecture is a split-cache consisting of a direct-mapped cache with small block sire and a fully-associative buffer with large block size. To overcome the disadvantage of small cache space, two mechanisms are enhanced by considering operational behaviors of multimedia applications: an adaptive multi-block prefetching to initiate various fetch sizes and an efficient block filtering to remove rarely reused data. The simulations on MediaBench show that the proposed 5KB-cache can provide equivalent performance and reduce energy consumption up to 40% as compared with 16KB 4-way set associative cache.