• Title/Summary/Keyword: gDNA

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High Frequency of TP53 but not K-ras Gene Mutations in Bolivian Patients with Gallbladder Cancer

  • Asai, Takao;Loza, Ernesto;Roig, Guido Villa-Gomez;Ajioka, Yoichi;Tsuchiya, Yasuo;Yamamoto, Masaharu;Nakamura, Kazutoshi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5449-5454
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    • 2014
  • Although genetic characteristics are considered to be a factor influencing the geographic variation in the prevalence of gallbladder cancer (GBC), they have not been well studied in Bolivia, which has a high prevalence rate of GBC. The purpose of this study was to examine the frequency of TP53 and K-ras mutations in Bolivian patients with GBC and to compare them with our previous data obtained in other high-GBC-prevalence countries, namely Japan, Chile, and Hungary. DNA was extracted from cancer sites in paraffin-embedded tissue from 36 patients using a microdissection technique. TP53 mutations at exons 5 to 8 and K-ras mutations at codons 12, 13 and 61 were examined using direct sequencing techniques. The data obtained were compared with those in the other high-GBC-prevalence countries. Of the 36 patients, 18 (50.0%) had a TP53 mutation (one mutation in each of 17 patients and three mutations in one patient), and only one (2.8%) had a K-ras mutation. Of the 20 TP53 mutations, 12 were of the transition type (60.0%). This rate was significantly lower than that in Chile (12/12, P<0.05). In addition, three mutations were of the CpG transition type (15.0%), which is a feature of endogenous mutation. All three were found in the hot spot region of the TP53 gene. In contrast, G:C to T:A transversion was found in Bolivia, suggesting the presence of exogenous carcinogens. Our findings suggest that the development of GBC in Bolivia is associated with both exogenous carcinogens and endogenous mechanisms. The identification of an environmental risk factor for GBC is needed to confirm these findings.

The DNA Repair Gene ERCC6 rs1917799 Polymorphism is Associated with Gastric Cancer Risk in Chinese

  • Liu, Jing-Wei;He, Cai-Yun;Sun, Li-Ping;Xu, Qian;Xing, Cheng-Zhong;Yuan, Yuan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6103-6108
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    • 2013
  • Objective: Excision repair cross-complementing group 6 (ERCC6) is a major component of the nucleotide excision repair pathway that plays an important role in maintaining genomic stability and integrity. Several recent studies suggested a link of ERCC6 polymorphisms with susceptibility to various cancers. However, the relation of ERCC6 polymorphism with gastric cancer (GC) risk remains elusive. In this sex- and age-matched case-control study including 402 GC cases and 804 cancer-free controls, we aimed to investigate the association between a potentially functional polymorphism (rs1917799 T>G) in the ERCC6 regulatory region and GC risk. Methods: The genotypes of rs1917799 were determined by Sequenom MassARRAY platform and the status of Helicobacter pylori infection was detected by enzyme-linked immunosorbent assay. Odd ratios (ORs) and 95% confidential interval (CI) were calculated by logistic regression analysis. Results: Compared with the common TT genotype, the ERCC6 rs1917799 GG genotype was associated with increased GC risk (adjusted OR=1.46, 95%CI: 1.03-2.08, P=0.035). When compared with (GT+TT) genotypes, the GG genotype also demonstrated a statistical association with increased GC risk (adjusted OR=1.38, 95%CI: 1.01-1.89, P=0.044). This was also observed for the male subpopulation (GG vs. TT: adjusted OR=1.71, 95%CI: 1.12-2.62, P=0.013; G allele vs. T allele: adjusted OR=1.32, 95%CI: 1.07-1.62, P=0.009). Genetic effects on increased GC risk tended to be enhanced by H. pylori infection, smoking and drinking, but their interaction effects on GC risk did not reach statistical significance. Conclusions: ERCC6 rs1917799 GG genotype might be associated with increased GC risk in Chinese, especially in males.

Characterization of a new cultivar of Auricularia auricula-judae 'Yong-A' (목이 신품종 '용아'의 특성)

  • Kim, Kil-Ja;Kim, Dami;Lee, Sook-Jae;An, Ho-Sub;Kim, Dong-Kwan;Kwon, Oh-Do
    • Journal of Mushroom
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    • v.16 no.3
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    • pp.198-202
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    • 2018
  • 'Yong-A' was bred by Jeollanamdo Agricultural Research and Extension Services in 2015. It was primarily selected through monospore crossing between JNM21008 and JNM21013 in 2009. The strain is named as JNM-Mi-194 and 'Yong-A', showing major characteristics with good production yield. The cultivation characteristics, yield potential, and fruiting body characteristics of 'Yong-A' are as follows; The wrinkle of the fruiting body was of branching type its shape was wave type. MCM, YM, Malt, and PDA media were suitable for growth of this cultivar. The number of effective stipes was 13 ea/0.9 kg. The minor axis of pileus was 5 cm and the major axis was 9 cm. The yield was 291 g per plastic bag (0.9 kg). Yong-A required 60 days for mycelial running at $20^{\circ}C$. The growth and primordial period required 24 days, which was shorter than that of JNM-Mi-21002 (the conventional cultivar). Somatic incompatibility was observed between parental lines and Yong-A. Analysis of genetic diversity in the new variety 'Yong-A' showed a different profile compared to that of the parent strain when Random Amplified Polymorphic DNA primers were used.

Isolation and characterization of a novel gossypol-degrading bacteria Bacillus subtilis strain Rumen Bacillus Subtilis

  • Zhang, Yunhua;Zhang, Zhengyou;Dai, Li;Liu, Ying;Cheng, Maoji;Chen, Lijuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.63-70
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    • 2018
  • Objective: The aim of the study was to isolate gossypol-degrading bacteria and to assess its potential for gossypol degradation. Methods: Rumen liquid was collected from fistulated cows grazing the experimental pasture. Approximately 1 mL of the rumen liquid was spread onto basal medium plates containing 2 g/L gossypol as the only source of carbon and was then cultured at $39^{\circ}C$ to isolate gossypol-degrading bacteria. The isolated colonies were cultured for 6 h and then their size and shape observed by microscope and scanning electron microscope. The 16S rRNA gene of isolated colonies was sequenced and aligned using National Center for Biotechnology Information-Basic Local Alignment Search Tool. The various fermentation conditions, initial pH, incubation temperature, inoculum level and fermentationperiod were analyzed in cottonseed meal (CSM). The crude protein (CP), total gossypol (TG), and free gossypol (FG) were determined in CSM after fermentation with isolated strain at $39^{\circ}C$ for 72 h. Results: Screening results showed that a single bacterial isolate, named Rumen Bacillus Subtilis (RBS), could use gossypol as a carbon source. The bacterium was identified by 16S rDNA sequencing as being 98% homologous to the sequence of Bacillus subtilis strain GH38. The optimum fermentation conditions were found to be 72 h, $39^{\circ}C$, pH 6.5, moisture 50%, inoculum level $10^7cell/g$. In the optimum fermentation conditions, the FG and TG content in fermented CSM decreased 78.86% and 49% relative to the control. The content of CP and the essential amino acids of the fermented CSM increased respectively, compared with the control. Conclusion: The isolation of a gossypol-degrading bacterium from the cow rumen is of great importance for gossypol biodegradation and may be a valuable potential source for gossypol-degradation of CSM.

Association of Single Nucleotide Polymorphism (SNP) in the PGK 2 Gene with Growth Traits in Pigs (돼지 PGK 2 유전자의 단일염기다형성 및 성장 형질과의 연관성 구명)

  • Jang, Hong-Chul;Kim, Sang-Wook;Lim, Da-Jeong;Kim, Jae-Young;Cho, Kyu-Ho;Kim, Myung-Jick;Lee, Ji-Woong;Choi, Bong-Hwan;Kim, Tae-Hun
    • Journal of Animal Science and Technology
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    • v.53 no.1
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    • pp.15-22
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    • 2011
  • The purpose of this study was to analyse of association between growth traits and single nucleotide polymorphisms (SNPs) polymorphism of phosphoglycerate kinase 2 (PGK 2) gene in pigs. The birth weight of piglet influences on weaning weight and survival rate that are import economic traits in pig industry. Also, these growth traits are representative factor to decrease a period getting to marketing weight as well as growth rate in pig. The PGK 2 is an isozyme that catalyzes the first ATP-generating step in the glycolytic pathwayand important enzyme related with energy metabolism. Twenty of SNPs were discoveredby genome structure analysis that compares the sequence on promoter and transcription region of PGK 2 gene in porcine chromosome 7. An association between PGK 2 SNPs and growth traits was analyzed in $F_2$ reciprocal-crossbred population between korean native pig (KNP) and Landrace. Association analysis indicated that polymorphism of the PGK 2 gene promoter region has significant effects on weight at birth (p<0.01) and weight at 3 weeks of age (p<0.0001). These results suggest that PGK 2 gene polymorphism was associated with energy metabolism and physiological function of growth in pig.

Construction and Production of Concatameric Human TNF Receptor-Immunoglobulin Fusion Proteins

  • Yim, Su-Bin;Chung, Yong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.81-89
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    • 2004
  • Tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and lymphotoxin-$\alpha$ (LT-$\alpha$, TNF-$\beta$) can initiate and perpetuate human diseases such as multiple sclerosis (MS), rheumatoid arthritis (RA), and insulin-dependent diabetes mellitus (IDDM). TNFs can be blocked by the use of soluble TNF receptors. However, since monomeric soluble receptors generally exhibit low affinity or function as agonists, the use of monomeric soluble receptors has been limited in the case of cytokines such as TNF-$\alpha$, TNF-$\alpha$, interleukin (IL)-1, IL-4, IL-6, and IL-13, which have adapted to a multi component receptor system. For these reasons, very high-affinity inhibitors were created for the purpose of a TNFs antagonist to bind the TNFR and trigger cellular signal by using the multistep polymerase chain reaction method. First, recombinant simple TNFR-Ig fusion proteins were constructed from the cDNA sequences encoding the extracellular domain of the human p55 TNFR (CD120a) and the human p75 TNFR (CD120b), which were linked to hinge and constant regions of human $IgG_1$ heavy chain, respectively using complementary primers (CP) encoding the complementary sequences. Then, concatameric TNFR-Ig fusion proteins were constructed using recombinant PCR and a complementary primer base of recombinant simple TNFR-Ig fusion proteins. For high level expression of recombinant fusion proteins, Chinese hamster ovary (CHO) cells were used with a retroviral expression system. The transfected cells produced the simple concatameric TNFR-Ig fusion proteins capable of binding TNF and inactivating it. These soluble versions of simple concantameric TNFR-Ig fusion proteins gave rise to multiple forms such as simple dimers and concatameric homodimers. Simple TNFR-1g fusion proteins were shown to have much more reduced TNF inhibitory activity than concatameric TNFR-Ig fusion proteins. Concatameric TNFR-Ig fusion proteins showed higher affinity than simple TNFR-Ig fusion proteins in a receptor inhibitor binding assay (RIBA). Additionally, concatameric TNFR-Ig fusion proteins were shown to have a progressive effect as a TNF inhibitor compared to the simple TNFR-Ig fusion proteins and conventional TNFR-Fc in cytotoxicity assays, and showed the same results for collagen induced arthritis (CIA) in mice in vivo.

Molecular Mechanism of Tetrabromobisphenol A (TBBPA)-induced Target Organ Toxicity in Sprague-Dawley Male Rats

  • Choi, Jae-Seok;Lee, Young-Jun;Kim, Tae-Hyung;Lim, Hyun-Jung;Ahn, Mee-Young;Kwack, Seung-Jun;Kang, Tae-Seok;Park, Kui-Lea;Lee, Jae-Won;Kim, Nam-Deuk;Jeong, Tae-Cheon;Kim, Sang-Geum;Jeong, Hye-Gwang;Lee, Byung-Mu;Kim, Hyung-Sik
    • Toxicological Research
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    • v.27 no.2
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    • pp.61-70
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    • 2011
  • Brominated flame retardants (BFRs) are present in many consumer products ranging from fabrics to plastics and electronics. Wide use of flame retardants can pose an environmental hazard, which makes it important to determine the mechanism of their toxicity. In the present study, dose-dependent toxicity of tetrabromobisphenol A (TBBPA), a flame retardant, was examined in male prepubertal rats (postnatal day 18) treated orally with TBBPA at 0, 125, 250 or 500 mg/kg for 30 days. There were no differences in body weight gain between the control and TBBPA-treated groups. However, absolute and relative liver weights were significantly increased in high dose of TBBPA-treated groups. TBBPA treatment led to significant induction of CYP2B1 and constitutive androstane receptor (CAR) expression in the liver. In addition, serum thyroxin (T4) concentration was significantly reduced in the TBBPA treated group. These results indicate that repeated exposure to TBBPA induces drug-metabolising enzymes in rats through the CAR signaling pathway. In particular, TBBPA efficiently produced reactive oxygen species (ROS) through CYP2B1 induction in rats. We measured 8-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of DNA oxidative damage, in the kidney, liver and testes of rats following TBBPA treatment. As expected, TBBPA strongly induced the production of 8-OHdG in the testis and kidney. These observations suggest that TBBPA-induced target organ toxicity may be due to ROS produced by metabolism of TBBPA in Sprague-Dawley rats.

Molecular Typing of Legionella pneumophila Isolated in Busan, Using PFGE (부산지역에서 분리한 레지오넬라균에 대한 PFGE를 이용한 molecular typing)

  • Park Eun-Hee;Kim Mi-Hee;Kim Joung-A;Han Nan-Sook;Lee Ju Hyeoun;Min Sang Gi;Park Yon Koung;Jin Seong Hyun;Jeong Gu Young;Bin Jae Hun
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.161-168
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    • 2005
  • In this study, we did the molecular typing of 39 environmental Legionella pneumophila serogroup 1 isolates collected from 2001-2003 in Busan using the pulsed-filed gel electrophoresis (PFGE). PFGE of SfiI fragments were divided into 10 pulsotypes $(A\~J)$, corresponding to $<65\%$ similarity and a subtype within each pulsotype was characterized by $>84\%$ similarity. The major cluster was pulsotype E $(46.2\%)$, which included 18 isolates and was divided into 4 subtypes $(E1\~E4)$. PFGE of NotI fragments were divided into 8 pulsotypes $(a\~h)$, corresponding to $<60\%$ similarity and a subtype within each pulsotype was characterized by $100\%$ similarity. The major cluster was pulsotype f $(38.5\%)$, which included 15 isolates. The ATCC type strain L. pneumophila serogroup 1 was identified as a different molecular pulsotype compare to the Busan isolates. It is possible that L. pneumophila serogroup 1 isolated in Busan with specific DNA pattern is comparable with those isolation in other cities in Korea.

Effects of Tillage on Organic Matters and Microbial Communities in Organically Cultivated Corn Field Soils (유기농 옥수수밭에서 경운이 토양 유기물 함량 및 미생물군집에 미치는 영향)

  • Ahn, Dalrae;An, Nan-Hee;Kim, Da-Hye;Han, Byeong-Hak;You, Jaehong;Park, InCheol;Ahn, Jae-Hyung
    • Korean Journal of Environmental Agriculture
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    • v.39 no.1
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    • pp.65-74
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    • 2020
  • BACKGROUND: Soil carbon sequestration has been investigated for a long time because of its potential to mitigate the greenhouse effect. No- or reduced tillage, crop rotations, or cover crops have been investigated and practiced to sequester carbon in soils but the roles of soil biota, particularly microorganisms, have been mostly ignored although they affect the amount and stability of soil organic matters. METHODS AND RESULTS: In this study we analyzed the organic matter and microbial community in organically cultivated corn field soils where no-tillage (NT) or conventional tillage (CT) had been practiced for about three years. The amounts of organic matter and recalcitrant carbon pool were 18.3 g/kg dry soil and 4.1 g C/kg dry soil, respectively in NT soils, while they were 12.4 and 2.5, respectively in CT soils. The amounts of RNA and DNA, and the copy numbers of bacterial 16S rRNA genes and fungal ITS sequences were higher in NT soils than in CT soils. No-tillage treatment increased the diversities of soil bacterial and fungal communities and clearly shifted the bacterial and fungal community structures. In NT soils the relative abundances of bacterial phyla known as copiotrophs, Betaproteobacteria and Bacteroidetes, increased while those known as oligotrophs, Acidobacteria and Verrucomicrobia, decreased compared to CT soils. The relative abundance of a fungal phylum, Glomeromycota, whose members are known as arbuscular mycorrhizal fungi, was about two time higher in NT soils than in CT soils, suggesting that the higher amount of organic matter in NT soils is related to its abundance. CONCLUSION: This study shows that no-tillage treatment greatly affects soil microbial abundance and community structure, which may affect the amount and stability of soil organic matter.

A Case of Neonatal Intrahepatic Cholestasis Caused by Citrin Deficiency (NICCD) Confirmed by SLC25A13 Mutation

  • Son, Yeong-Bae;Jang, Ju-Yeong;Park, Hyeong-Du;Lee, Su-Yeon
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.14 no.2
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    • pp.186-190
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    • 2014
  • Citrullinemia type 2 (citrin deficiency) is an autosomal recessive inborn error metabolism, caused by the SLC25A13 gene mutation. Citrin deficiency is associated with two clinical phenotype; neonatal-onset type II citrullinemia (CTLN2), also known as neonatal intraphepatic cholestasis caused by citrin deficiency (NICCD) and adult-onset CTLN2. Clinical manifestations of NICCD include poor growth, intrahepatic cholestasis, liver dysfunction and increased plasma citrulline, methionine, threonine, arginine. The molecular diagnosis could be confirmed by SLC25A13 gene mutation analysis. A 3-month-old male infant with persistent jaundice was referred for evaluation. Newborn screening was normal at birth. Mild elevation of serum ammonia and AST/ALT were observed. Plasma amino acid analysis showed significantly elevated citrulline, methionine, threonine. DNA sequence analysis of the SLC25A13 gene revealed two compound heterozygous mutations, c.[852_855del]($p.Met285Profs^*2$) and [1180+1G>A]. We suggest that NICCD should be considered as one of the cause of in infants with cholestatic jaundice, although the newborn screening was normal.