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Association of a Newly Identified Variant of DNA Polymerase Beta (polβΔ63-123, 208-304) with the Risk Factor of Ovarian Carcinoma in India

  • Khanra, Kalyani;Bhattacharya, Chandan;Bhattacharyya, Nandan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.1999-2002
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    • 2012
  • Background: DNA polymerase is a single-copy gene that is considered to be part of the DNA repair machinery in mammalian cells. The encoded enzyme is a key to the base excision repair (BER) pathway. It is evident that pol beta has mutations in various cancer samples, but little is known about ovarian cancer. Aim: Identification of any variant form of $pol{\beta}$ cDNA in ovarian carcinoma and determination of association between the polymorphism and ovarian cancer risk in Indian patients. We used 152 samples to isolate and perform RT-PCR and sequencing. Results: A variant of polymerase beta (deletion of exon 4-6 and 11-13, comprising of amino acid 63-123, and 208-304) is detected in heterozygous condition. The product size of this variant is 532 bp while wild type pol beta is 1 kb. Our study of association between the variant and the endometrioid type shows that it is a statistically significant factor for ovarian cancer [OR=31.9 (4.12-246.25) with p<0.001]. The association between variant and stage IV patients further indicated risk (${\chi}^2$ value of 29.7, and OR value 6.77 with 95% CI values 3.3-13.86). The correlation study also confirms the association data (Pearson correlation values for variant/stage IV and variant/endometrioid of 0.44 and 0.39). Conclusion: Individuals from this part of India with this type of variant may be at risk of stage IV, endometrioid type ovarian carcinoma.

Identification and Analysis of Putative Polyhydroxyalkanoate Synthase (PhaC) in Pseudomonas fluorescens

  • Lim, Ju Hyoung;Rhie, Ho-Gun;Kim, Jeong Nam
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1133-1140
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    • 2018
  • Pseudomonas fluorescens KLR101 was found to be capable of producing polyhydroxyalkanoate (PHA) using various sugars and fatty acids with carbon numbers ranging from 2 to 6. The PHA granules consisted mainly of a poly(3-hydroxybutyrate) homopolymer and/or poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer. Genomic DNA of P. fluorescens was fractionated and cloned into a lambda library, in which a 5.8-kb fragment that hybridized to a heterologous phaC probe from Ralstonia eutropha was identified. In vivo expression in Klebsiella aerogenes KC2671 (pUMS), restriction mapping, Southern hybridization experiments, and sequencing data revealed that PHA biosynthesis by P. fluorescens relied upon a polypeptide encoded by a 1,683-bp non-operonal ORF, which was preceded by a possible -24/-12 promoter and highly similar to DNA sequences of a gene encoding PHA synthase in the genus Pseudomonas. In vivo expression of the putative PHA synthase gene ($phaC_{Pf}$) in a recombinant Escherichia coli strain was investigated by using glucose and decanoate as substrates. E. coli (${phaC_{Pf}}^+$, pUMS) grown in medium containing glucose accumulated PHA granules consisting mainly of 3-hydroxybutyrate, whereas only a trace amount of 3-hydroxydecanoate was detected from an E. coli fadR mutant (${phaC_{Pf}}^+$) grown in medium containing decanoate. In vitro enzymatic assessment experiments showed that 3-hydroxybutyryl-CoA was efficiently used as a substrate of purified $PhaC_{Pf}$, suggesting that the putative PHA synthase of P. fluorescens utilizes mainly short-chain-length PHA precursors as a substrate.

Rapid diagnosis of bovine tuberculosis in slaughter cattle using PCR (PCR 기법을 이용한 도축 소의 결핵병 신속진단)

  • Koh, Ba-Ra-Da;Kim, Hyun-Joong;Park, Duk-Woong;Park, Seong-Do;Kim, Jae-Ik;Park, Jong-Tae;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.393-406
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    • 2007
  • Bovine tuberculosis is an important zoonosis worldwide. Mycobacterium bovis, the causative agent of this disease in cattle, is also a pathogen for humans and several economically important animals. The cases of tuberculosis are reported in two cow found at slaughter house located in Gwangju city. Histopathologically, in the lymph nodes, granulomas consisted of large areas of necrosis surrounded by variable thick bands of cellular infiltrate containing macrophages, Langhans-type multinucleated giant cells and lymphocytes. Lesions in the lung followed the same developmental pattern as did lesions in the lymph nodes with some exceptions. With the acid-fast staining, numerous mycobacteria were revealed in the lung and lymph nodes. M bovis was confirmed as a causative agent in these cattle using bacterial isolation and PCR and restriction fragment length polymorphism method based on a unique 12.7 kb fragment insertion sequence from the Mycobacterium tuberculosis genome and the pncA polymorphism, The insertion element IS6110 and IS1081 were present M bovis isolated from lungs and lymph nodes of cattle using PCR assay. These cases are interesting and important in public health aspect that M bovis-infected cattle were found during a routine post-mortem inspection at slaughter house.

Comparison of Green Color Retention of Zoysiagrass and Cool-season Grass under Multilayer System, USGA System, and Mono-layer System of Sports Field (스포츠용 다단구조, USGA구조 및 약식구조 지반에서 한국잔디 및 한지형 잔디의 녹색기간 비교)

  • Kim, Kyoung-Nam
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.342-353
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    • 2016
  • This study was initiated to evaluate green color retention under three different soil systems. Several turfgrasses were evaluated in multi-layer, USGA, and mono-layer systems. Turfgrass entries were comprised of three cultivars of Korean lawngrass (Zoysia japonica Steud.) as warm-season grass (WSG) and three blends and three mixtures of Kentucky bluegrass (KB, Poa pratensis L.), perennial ryegrass (PR, Lolium perenne L.), and tall fescue (TF, Festuca arundinacea Schreb.) as cool-season grass (CSG). Significant differences were observed in visual turf color and green color retention among soil systems and turfgrasses. Both the multi-layer and USGA systems were highly associated with better color ratings and longer color retention, as compared with the mono-layer system. Seasonal variation of visual turf color greatly occurred from late December to early spring. CSG exhibited longer color retention than did WSG. The latter maintained green color for approximately 6 months, regardless of the soil system. Spring green-up of Korean lawngrass occurred from early to middle May, while it underwent discoloration from late October to early November. Among the CSGs green-up occurred between early March and early April and leaf color was maintained until middle December to early February. Therefore, the CSGs were green for 8.5 to 11 months, depending on turfgrass and soil system. The mean period of green color duration across all soil systems was approximately 10-11, 9-10 and 8.5-9.0 months for PR, KB and TF, respectively. As for the CSG mixtures, the greater the proportion of PR, the longer the green color retention, while the higher the proportion of TF, the shorter the color retention. There was greater variation in green color duration among the CSGs than the WSGs. Across soil systems, color retention differences of 2 to 6 days were observed for the Korean lawngrass, but 7 to 36 days for the CSGs. These results demonstrate that green color retention varied greatly according to soil systems and also among turfgrasses. Selections of turfgrass and soil system should be made using a concept-oriented approach, when establishing garden, park, soccer field, golf course and other sports field. Information obtained in this study can be used to select soil systems and turfgrasses based on the expected degree of leaf color retention.

A Comparative Study on the Graphics Usage by Terrestrial Broadcasters TV News - Focusing on the Evening Main News of KBS, MBC, and SBS - (지상파방송사 TV뉴스의 그래픽 현황 비교 연구 -KBS, MBC, SBS 저녁 메인뉴스 중심으로-)

  • Kim, Sang-cheol;Kim, Kwang-Ho
    • Journal of Digital Contents Society
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    • v.16 no.3
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    • pp.397-406
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    • 2015
  • Terrestrial broadcasters are devoting great efforts in order to increase sense of reality that would help viewers feel as if they were watching events on site by producing-depth, differentiated news and offering a variety of information to them. They maximize news video graphic effects by replacing news set and install large display units, rail camera, and Jimmy Jib camera. This study aims to grasp whether there is any difference among the main evening news of the three major terrestrial broadcasters. KBS, MBC, and SBS used 18%, 15%, and 12% of graphic usage hours respectively while the number of graphic usage cases amounted to 267, 329, and 276 for each of them. The number of cases where in-depth news report hours exceeded three minutes that require increasingly more virtual reality graphic technology production was 16 for KBS, 7 for MBC, and 5 for SBS. MBC showed the highest number of uses in graphics while KBS showed more graphic use hours than the other two. There have been a number of preceding studies on news contents, but it is significant that this study looked at how much graphics technologies are used in such news video contents.

On Reducing the Shadow Region for Extending the Service Area of TBN-Jeonju Broadcasting (전주교통방송 가청권 확대를 위한 음영 지역 해소 방안)

  • Kim, Tae-Hoon;Lee, Moon-Ho;Lee, Mi-Sung;Cho, Kye-Mun
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.21 no.12
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    • pp.1352-1370
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    • 2010
  • In this paper, we investigated the shadow area of radio programs which are currently being broadcast via FM 102.5 MHz in Jeollabuk-do consisting of 6 cities and 8-guns. The field strength data of FM 102.5 MHz was used with criterion of shadow area. As a result, the field strength of the eastern mountainous area in Jeollabuk-do was very low. We also measured, compared and analyzed the field strength of 102.5 MHz(TBN-Jeonju), 100.7 MHz(KBS) and 99.1 MHz (MBC) in Jeollabuk-do. Based on the field strength data, we tried to find what correlation exists and how much influence is between difference of antenna output power and solution of shadow area. In result, enhancement of antenna output power is not solution in terms of reducing the shadow Area of 102.5 MHz(TBN-Jeonju). In addition, we measured, compared and analyzed the field strength of 102.5 MHz(TBN-Jeonju) and 106.9 MHz(EBS). In fact, the relay stations with 104.1 MHz, 104.9 MHz, 105.3 MHz and 107.5 MHz of EBS FM has a number of FM relay stations in the eastern mountainous area of Jeollabuk-do. As a result, we confirmed that EBS almost did not have shadow area in the eastern mountainous area of Jeollabuk-do based on the field strength data. As in the case of EBS and KBS, we suggest installing TBN-Jeonju FM relay station with the best solution to reducing the shadow area to extend the audible range of TBN-Jeonju.

Petorshemical Study on the Mantle Xwnoliths in alkli basalts from S. Korea: P-T Regime of Upper Mantle (남한의 알카리 현무암에 분포하는 맨틀포획암의 암석화학적 연구: 상부맨틀포획암의 암석화학적 연구: 상부맨틀의 온도 및 압력 추정)

  • 이한영
    • The Journal of the Petrological Society of Korea
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    • v.4 no.2
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    • pp.104-123
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    • 1995
  • Mantle xenoliths in alkali basalt from Boun, Gansung area, and Baegryung island in S. Korea are spinel lherzolites composed of olivine, orthopyroxene, clinopyroxene, and spinel. The xenoliths generally show triple junctions among grams, kink-banding in olivine and pyroxenes, and protogranular and eqigranular textures having m orlentatron of specific direction. Anhedral brown spinels are disseminated in the intergranular spaces of minerals. Mineral compositions are very homogeneous without compositional zonation from rim to core in grains regardless different locahties. Olivine shows Fo. component of 89.0-90.2 and low CaO of 0.03-0.12wt%, orthopyroxene is enstatite with En component of 89.0 - 90.0 and $Al_2O_3$ of 4-5wt%, and clinopyroxene is diopside having En. component of 47.2-49.1 and $Al_2O_3$ of 7.42-7.64wt% from Boun and 4.70-4.91wt% from Baegryung showing local variation. Spinel shows the distinctive negative trend with increasing of A1 and decreasing of Cr, and Mg value and Cr number are 75.1-81.9 and 8.5-12.6, respectively. To estlmate T and P for these mantle xenoliths pyroxene-geothermometers (Wood and Banno, 1973; Wells, 1977; Mercier, 1980; Sachtleben and Seck, 1981; Bertrand and Mercier, 1985; Brey and Kohler, 1990) and Al-solubility geobarometer (Mercier, 1980; Lane and Ganguly, 1980) are used. Temperatures of Mercier (1980) and Sachtleben and Seck (1981) are compatible and equilibrium temperatures of xenoliths, average value of these two, aiie from $970^{\circ}C$ to $1020^{\circ}C$, and equihbrium pressures derived from Mercier (1980) are in the range of 12-19 Kb (42-63 Km). These temperatures and pressures seem to be reasonble wlth the consideration of Al-isopleths in MAS system (Lane and Ganguly, 1980) and Fe effect on Al-solubility in orthopyroxene (Lee and Ganguly, 1988). Equllibrium of temperatures and pressures of xenoliths in P-T space belong to ocenanic geothem among the Mercier's mantle geotherms (1980) and are completely different from continental geotherms of S. Africa (Lesotho) and S. India having different geologcal ages. anera1 compositions of spmel-lherzohtes in S. Korea and eastern China are primitwe and paleogeothems of both are very s~mllar, but degrees of depletion of upper mantle could be locally different from each other since eastern China has various depleted xenoliths due to different degrees of partial melting.

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Cloning and Expression of the Cathepsin F-like Cysteine Protease Gene in Escherichia coli and Its Characterization

  • Joo, Han-Seung;Koo, Kwang-Bon;Park, Kyun-In;Bae, Song-Hwan;Yun, Jong-Won;Chang, Chung-Soon;Choi, Jang-Won
    • Journal of Microbiology
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    • v.45 no.2
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    • pp.158-167
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    • 2007
  • In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

Anti-Angiogenic and Anti-Cell Adhesion Effect of the Camellia japonica Flower Extract (동백꽃 추출물의 신생혈관생성 및 세포부착 억제 효과)

  • Heo, In-Do;Seo, Hyo-Jin;Kim, Jong-Deog
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1152-1156
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    • 2007
  • The Camella japonica flower(CJF) extract was studied for their anti-angiogenenic and anti-cell adhesion effect. CJF-extract inhibited the tube formation on human umbilical vein endotherial cells(HUVEC) with butanol extract by 70.2%, acetone extract by 54.2%, ethyl acetate extract by 37.0%, chloroform extract by 21.2%. Cell adhesion molecules were effectively suppressed at different concentration of CJF at 50, 100, 200 ug/well such as for intercellular adhesion molecule(ICAM) by 5.9%, 29.4% and 52.9%, for vascular cell adhesion molecule(VCAM) by 12.5%, 43.8% and 62.5%, for E-selectin by 7.1%, 21.4% and 35.7%, respectively. Signal molecules of vascular endotherial growth factor receptor 2(VEGFR2), ${/beta}$-catenin and PI3K are inhibited by different concentration of CJF at 10, 20 and 30 ${\mu}g/mL$ with western blot. Angiogenesis will be inhibited with suppressing NF-kB molecule resulted in signal molecules blocked by CJF. CJF will be useful materials for treatment of angiogenesis related diseases such as cancer, metastasis, rheumathioid arthritis and obesity.

Gene Expression Analysis of Inducible cAMP Early Repressor (ICER) Gene in Longissimus dorsi of High- and Low Marbled Hanwoo Steers (한우 등심부위 근육 내 조지방함량에 따른 inducible cAMP early repressor (ICER) 유전자발현 분석)

  • Lee, Seung-Hwan;Kim, Nam-Kuk;Kim, Sung-Kon;Cho, Yong-Min;Yoon, Du-hak;Oh, Sung-Jong;Im, Seok-Ki;Park, Eung-Woo
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1090-1095
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    • 2008
  • Marbling (intramuscular fat) is an important factor in determining meat quality in Korean beef market. A grain based finishing system for improving marbling leads to inefficient meat production due to an excessive fat production. Identification of intramuscular fat-specific gene might be achieved more targeted meat production through alternative genetic improvement program such as marker assisted selection (MAS). We carried out ddRT-PCR in 12 and 27 month old Hanwoo steers and detected 300 bp PCR product of the inducible cAMP early repressor (ICER) gene, showing highly gene expression in 27 months old. A 1.5 kb sequence was re-sequenced using primer designed base on the Hanwoo EST sequence. We then predicted the open reading frame (ORF) of ICER gene in ORF finder web program. Tissue distribution of ICER gene expression was analysed in eight Hanwoo tissue using realtime PCR analysis. The highest ICER gene expression showed in Small intestine followed by Longissimus dorsi. Interestingly, the ICER gene expressed 2.5 time higher in longissimus dorsi than in same muscle type, Rump. For gene expression analysis in high- and low marbled individuals, we selected 4 and 3 animal based on the muscle crude fat contents (high is 17-32%, low is 6-7% of crude fat contents). The ICER gene expression was analysed using ANOVA model. Marbling (muscle crude fat contents) was affected by ICER gene (P=0.012). Particularly, the ICER gene expression was 4 times higher in high group (n=4) than low group (n=3). Therefore, ICER gene might be a functional candidate gene related to marbling in Hanwoo.