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Comparison of Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, and Loop-Mediated Isothermal Amplification for the Detection of Cronobacter sakazakii in Milk Powder (분유에 오염된 Cronobacter sakazakii 검출을 위한 중합효소연쇄반응, 실시간중합효소연쇄반응, 등온검출법의 비교)

  • Kim, Young-Joo;Seo, Sheungwoo;Wang, Xiaoyu;Seo, Dong Joo;Lee, Min Hwa;Son, Na Ry;Lee, Bog-Hieu;Choi, Changsun
    • Food Science of Animal Resources
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    • v.33 no.5
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    • pp.610-616
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    • 2013
  • Loop-mediated isothermal amplification (LAMP) is an emerging detection technology for the amplification of DNA under isothermal conditions. The aim of this study was to develop a rapid and reliable LAMP technique for the detection of Cronobacter sakazakii in milk powder. In order to enhance the sensitivity and specificity, LAMP primers targeting outer membrane protein A (ompA) gene of C. sakazakii were designed using Explorer V4 software. Thirty seven C. sakazakii strains and 13 pathogenic microorganisms were used for comparative detection of C. sakazakii using polymerase chain reaction (PCR), real-time PCR, and LAMP. LAMP developed in this study could specifically detect C. sakazakii strains without cross-reactivity with other foodborne pathogens. LAMP products amplified from ompA gene of C. sakazakii were digested with with HhaI and NruI enzyme. The specificity of LAMP was confirmed by restriction fragment length polymorphism (RFLP) analysis. LAMP could detect C. sakazakii within 1 h without bacterial culture and its detection limit was as low as 1 CFU/mL C. sakazakii in milk. In the comparison of the sensitivity, LAMP showed 10,000- and 100-times higher detection limit than PCR or real-time PCR, respectively. Therefore, this study can conclude that LAMP is a rapid and reliable detection technique for C. sakazakii contaminated in powdered milk.

Development and Evaluation of PCR-Based Detection for Pseudomonas syrinage pv. tomato in Tomato Seeds (토마토 종자로부터 PCR을 이용한 Pseudomonas syringae pv. tomato의 검출)

  • Cho, Jung-Hee;Yim, Kyu-Ock;Lee, Hyok-In;Yea, Mi-Chi;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.17 no.3
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    • pp.376-380
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    • 2011
  • The bacterial speck of tomato caused by Pseudomonas syringae pv. tomato leads to serious economic losses especially on fruits of susceptible genotype. Thus, Pseudomonas syringae pv. tomato is a plant quarantine bacterium in many countries including Korea. In this study, we developed specific PCR assays for detection of the bacterium from tomato seeds. A specific primer set is designed from the hrpZ gene for specific detection of Pseudomonas syringae pv. tomato. A 501 bp PCR product corresponding to hrpZ gene was amplified only form Pseudomonas syringae pv. tomato strains, but no PCR product was amplified from other tomato bacterial pathogens, such as Pseudomonas syringae pv. glycinea, P. syringae pv. maculicola, P. syringae pv. atropurpurea, P. syringae pv. morsprunorum, and from other P. syringae pathovar strains. The nested-PCR primer set corresponding to an internal fragment of the 501 bp sequence (hrpZ) gine was used to specific detection of Pseudomonas syringae pv. tomato in tomato seed. A 119 bp PCR product using nested PCR primer was highly specific and sensitive to detect low level of Pseudomonas syrigae pv. tomato in tomato seeds. We believe that the PCR assays developed in this study is very useful to detect Pseudomonas syringae pv. tomato from the tomato seeds.

Prediction of Rock Fragmentation and Design of Blasting Pattern based on 3-D Spatial Distribution of Rock Factor (발파암 계수의 3차원 공간 분포에 기초한 암석 파쇄도 예측 및 발파 패턴 설계)

  • Shim Hyun-Jin;Seo Jong-Seok;Ryu Dong-Woo
    • Tunnel and Underground Space
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    • v.15 no.4 s.57
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    • pp.264-274
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    • 2005
  • The optimum blasting pattern to excavate a quarry efficiently and economically can be determined based on the minimum production cost which is generally estimated according to rock fragmentation. Therefore it is a critical problem to predict fragment size distribution of blasted rocks over an entire quarry. By comparing various prediction models, it can be ascertained that the result obtained from Kuz-Ram model relatively coincides with that of field measurements. Kuz-Ram model uses the concept of rock factor to signify conditions of rock mass such as block size, rock jointing, strength and others. For the evaluation of total production cost, it is imperative to estimate 3-D spatial distribution of rock factor for the entire quarry. In this study, a sequential indicator simulation technique is adopted for estimation of spatial distribution of rock factor due to its higher reproducibility of spatial variability and distribution models than Kriging methods. Further, this can reduce the uncertainty of predictor using distribution information of sample data The entire quarry is classified into three types of rock mass and optimum blasting pattern is proposed for each type based on 3-D spatial distribution of rock factor. In addition, plane maps of rock factor distribution for each ground levels is provided to estimate production costs for each process and to make a plan for an optimum blasting pattern.

Effects of c.494A>C and c.267T>G SNPs in OCX-32 Gene of Korean Native Chicken on Egg Production Traits (OCX-32 유전자 내 c.494A>C 및 c.267T>G SNP이 한국 재래닭 산란형질에 미치는 효과 분석)

  • Lee, Ji-Yeon;Choi, So-Young;Kim, Chong-Dae;Hong, Yeong Ho;Jeong, Dong Kee;Lee, Sung-Jin
    • Korean Journal of Poultry Science
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    • v.41 no.3
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    • pp.191-196
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    • 2014
  • The identification and utilization of potential candidate genes with significant effects on economically important traits have become increasingly important in poultry breeding programs. The ovocalyxin-32 (OCX-32) gene is located chromosome 9 in chicken, plays an important role in eggshell formation. This study was performed to assess the association between single nucleotide polymorphisms (SNPs) of OCX-32 gene and egg production traits in the Korean native chicken. Four Korean native chicken population (n = 181; including 46 females of Ogol, 46 females of white, 43 females of gray and 46 females of black) were used to analyze two SNPs (c.494A>C and c.267T>G) in the OCX-32 gene by PCR-RFLP (Polymerase Chain Reaction-Restricted Fragment Length Polymorphism). We measured egg production traits of age at first egg, first egg weight, egg production ratio and egg weight. The SNP c.494A>C was significantly associated with egg production ratio in Korean Ogol chickens (p<0.001) and egg weight in Korean white chickens (p<0.05). SNP c.267T>C was significantly associated with egg weight in Korean Ogol chickens (p<0.05). But there was no significant association in Korean gray and black chickens. Results suggest the possibility of using molecular markers in OCX-32 gene as a tool for performance and egg production traits in Korean native chicken breeding program.

Association between Economic Traits and Candidate Gene Polymorphism in Korean Native Pig and Duroc (한국 재래 돼지와 듀록의 경제형질과 후보 유전자 다형성간의 연관성 분석)

  • Kim, M.J.;Oh, J.D.;Cho, G.H.;Lee, J.H.;Lee, S.S.;Hong, Y.S.;Jeon, K.J.;Jeon, G.J.;Lee, H.K.
    • Journal of Embryo Transfer
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    • v.21 no.4
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    • pp.273-280
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    • 2006
  • MC4R, PRKAG3, FABP3, and ESR have reported as important candidate genes related to some economic traits in pigs. To investigate the association between these genes and economic traits, the analysis of restriction fragment length polymorphism (RFLP) was conducted on 147 individuals (96 Durocs and 86 Korea native pigs; KNP) using single nucleotide polymorphism (SNP). Different genotype frequencies of 4 candidate genes were observed in Duroc and KNP. There were significant associations between MC4R polymorphic site and average daily gain (ADG, p<0.05) and backfat thickness (BF, p<0.05) in the Duroc, ADG (p<0.05) and days to 70 kg (p<0.05) in KNP. PRXAG3 polymorphic site were significantly .elated to BF (p<0.05) in the Duroc, ADG (p<0.05) and days to 70 kg (p<0.05) in the KNP. In FABP3, association with BF (p<0.05) in the Duroc, ADG (p<0.05) and days to 70 kg (p<0.05) in the KNP were found. ESR polymorphic site was not significantly associated to any other traits.

Prevalence of Aflatoxin Induced p53 Mutation at Codon 249 (R249s) in Hepatocellular Carcinoma Patients with and without Hepatitis B Surface Antigen (HBsAg)

  • Chittmittrapap, Salyavit;Chieochansin, Thaweesak;Chaiteerakij, Roongruedee;Treeprasertsuk, Sombat;Klaikaew, Naruemon;Tangkijvanich, Pisit;Komolmit, Piyawat;Poovorawan, Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7675-7679
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    • 2013
  • Background: A missense mutation in exon 7 (R249S) of the p53 tumor suppressor gene is characteristic of aflatoxin B1 (AFB1) exposure. AFB1 is believed to have a synergistic effect on hepatitis virus B (HBV) carcinogenesis. However, results of studies comparing R249S prevalence among patients are conflicting. The aim of this study was to determine the prevalence of the R249S mutation in hepatocellular carcinoma (HCC) patients with or without positive HBsAg. Materials and Methods: Paraffin embedded liver tissues were obtained from 124 HCC patients who underwent liver resection and liver biopsy in King Chulalongkorn Memorial Hospital. Restriction fragment length polymorphism (RFLP) was utilized to detect the R249S mutation. Positive results were confirmed by direct sequencing. Results: Sixty four (52%) patients were positive for HBsAg and 18 (15%) were anti-HCV positive. 12 specimens tested positive by RFLP. Ten HCC patients (8.1%) were confirmed to be R249S positive by Sanger sequencing (AGG to AGT). Out of these 10, six were HBsAg positive, and out of the remaining 4, two were anti-HCV positive. The R249S prevalence among HCC patients with positive HBsAg was 9.4% compared to 6.7% for HBsAg negative samples. Patients with the R249S mutation were younger ($55{\pm}10$ vs $60{\pm}13$ year-old) and tended to have a more advanced Edmonson-Steiner grade of HCC, although differences did not reach statistical significance. Conclusions: Our study shows moderate prevalence of aflatoxin B1-related p53 mutation (R249S) in HCC with or without HBsAg. HBsAg positive status was not associated with R249S prevalence.

Tactics and Pitfalls of MED(Micro Endoscopic Discectomy) System for Lumbar Disc - For Surgeons Who Wish to Attempt - (요추간반 탈출증에 있어 MED(Microendoscopic Discectomy) System을 이용한 수술의 효율성과 수기)

  • Hong, Hyun Jong;Oh, Seong Hoon;Bak, Kwang Hum;Kim, Jae Min;Kim, Choong Hyun;Kim, Young Soo;Ko, Yong;Oh, Suck Jun;Kim, Kwang Myung;Lee, Sang Gu;Kim, Nam Kyu
    • Journal of Korean Neurosurgical Society
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    • v.29 no.1
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    • pp.35-43
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    • 2000
  • Objective : Percutaneous lumbar approaches such as arthroscopic discectomy, laser discectomy, and nucleotome remain controversial and have technical limitations to free fragment disc, bony pathology and access to L5-S1, The purpose of this study was to determine efficacy of this new endoscopic system and to report techniques and tactics. Methods : From July 1997 to May 1998, we treated 40 consecutive patients(43 levels) with the MED system. Mean age was 32 years(range ; 18 to 62). There were 30 males and 10 females. All patients had sciatica with SLRT limitation. There were 23 patients with disc herniation at L4-5 and 14 patients at L5-S1. Three patients had 2 level disc herniations. There was one far lateral disc herniation at L4-5. Results : Using modified MacNab criteria, there were 37 excellent results and 3 good result. Most patients were discharged within 3-4 days except 2 patients with dural tearing. There were no other complications. Mean operation time was 1.5 hours(range : 40 minutes to 2.5 hours). Conclusion : The MED system is a reliable approach to lumbar disc herniations. This system combines the advantages of conventional open surgery and a minimally invasive technique. As tactics for the doctors who wish to attempt, "palpate" the lamina by first dilator, identification of interlaminar space by removal of overlying soft tissue and confirmation of the shoulder portion of nerve root before discectomy are important to this procedure. We conclude that lumbar disc herniations can be successfully treated with MED approach.

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Evaluation of Plasma D-Dimer Concentration in Dogs with Chronic Mitral Valve Insufficiency (만성 이첨판 폐쇄부전증에 걸린 개에서 혈장 D-dimer 농도 측정 연구)

  • Park, Joungsoon;Suh, Sang-Il;Oh, Yeonsu;Hyun, Changbaig
    • Journal of Veterinary Clinics
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    • v.32 no.1
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    • pp.5-8
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    • 2015
  • D-dimer is a fibrin degradation product (FDP), a small protein fragment present in the blood after a blood clot is degraded by fibrinolysis. D-dimer concentration is widely used for determining thrombosis and thromboembolism. Because one major cause of thromboembolism is congestive heart failure in human, we investigated the degree and risk of thromboembolism in dogs with different stage of congestive heart failure caused by chronic mitral valvular insufficiency (CMVI). The plasma level of d-dimer was evaluated in 20 healthy dogs and 30 dogs with different stage of congestive heart failure caused by CMVI. The d-dimer concentrations were measured by a commercialized assay kit. The plasma levels of d-dimer were not significantly different between healthy and CMVI dogs. Furthermore, there was no association of d-dimer concentrations to left atrium to aorta (LA/Ao) ratio, left ventricular dimension at diastole to aorta (LVIDd/Ao) ratio and severity of heart failure in our study population. Our study results implied that the degree of thromboembolism in canine heart failure might be minimal or the plasma d-dimer test might not be reliable for detecting thromboembolism in dogs.

CYP2C19 Genotype Could be a Predictive Factor for Aggressive Manifestations of Hepatocellular Carcinoma Related with Chronic Hepatitis B Infection in Thailand

  • Nun-anan, Pongjarat;Chonprasertsuk, Soonthorn;Siramolpiwat, Sith;Tangaroonsanti, Anupong;Bhanthumkomol, Patommatat;Pornthisarn, Bubpha;Vilaichone, Ratha-korn
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3253-3256
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    • 2015
  • Background: Chronic hepatitis B virus (HBV) infection related hepatocellular carcinoma (HCC) is a major health problem in the Asia-Pacific region including Thailand. Several factors have been proposed as contributing to hepatocarcinogenesis. This study was aimed to investigate the impact of CYP2C19 genotypic polymorphism in HCC related to chronic HBV infection in Thailand. Materials and Methods: A cross-sectional study was performed between April 2014 and January 2015. Chronic HBV patients with HCC (n=50) and without HCC (n=50) were included. Clinical information and blood samples of all patients were collected. The CYP2C19 genotype was determined by polymerase chain reaction-restriction fragment length polymorphism method, and was classified as rapid metabolizer (RM), intermediate metabolizer (IM) or poor metabolizer (PM). Results: The CYP2C19 genotype frequencies of RM, IM and PM in HBV patients were found to be 19/50 (38%), 25/50 (50%) and 6/50 (12%), respectively. The CYP2C19 genotype frequencies of RM, IM and PM in HBV with HCC patients were 21/50 (42%), 25/50 (50%) and 4/50 (8%), respectively. The distribution of CYP2C19 genotype was not different between patients with and without HCC. Interestingly, among HBV with HCC patients, the RM genotype of CYP2C19 tended to increase risk of aggressive manifestation (OR=2.89, 95%CI=0.76-11.25, P-value=0.07), compared with non RM genotype carriers. Conclusions: CYP2C19 genotype IM was the most common genotype in Thai patients with chronic HBV infection. In addition, genotype RM could be an associated factor for aggressive presentation in HCC related to chronic HBV infection.

Analysis of Chromosomal Aberration Induced by Low Dose of Radiation (저선량방사선에 의한 염색체이상 빈도)

  • Yi, Chun-Ja;Ha, Sung-Whan
    • Radiation Oncology Journal
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    • v.11 no.2
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    • pp.233-240
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    • 1993
  • Chromosomal aberration analysis, as a basis for biological radiation dosimetry, was performed for radiation dose ranges below 150 cGy. The yield, ratio of lymphocytes with dicentric and/or ring chromosomes, was 0, 0, 0.4, 0.5, 0.6, 0.8, 1.8, 5.5, 8.0, and $18.5\%$ for 0,5, 10, 15, 20, 25, 50, 75, 100 and 150 cGy, respectively. The Qdr, ratio of dicentric and ring chromosomes in total lymphocytes, was 0, 0, 0.004, 0.005, 0.006, 0.009, 0.018, 0.055, 0.084 and 0.207, respectively. The Qdr, ratio of dicentric and ring chromosomes in lymphocytes with aberration, was 1.0 for the radiation doses up to 75 cGy and 1.05 and 1.11 for 100 and 150 cGy, respectively. From the results, it seems possible to estimate radiation dose from Ydr when the exposure is 25 cGy or more. All the 5 radiation workers studied, with exposure much less than 1 mSv per month, had chromosomal aberrations. And acentric fragment pairs, in addition to dicentric and ring chromosomes, showed good dose response relationship and so may be useful for biological dosimetry for low dose radiation.

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