• Title/Summary/Keyword: Amplification Mechanism

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Spoilage Lactic Acid Bacteria in the Brewing Industry

  • Xu, Zhenbo;Luo, Yuting;Mao, Yuzhu;Peng, Ruixin;Chen, Jinxuan;Soteyome, Thanapop;Bai, Caiying;Chen, Ling;Liang, Yi;Su, Jianyu;Wang, Kan;Liu, Junyan;Kjellerup, Birthe V.
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.955-961
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    • 2020
  • Lactic acid bacteria (LAB) have caused many microbiological incidents in the brewing industry, resulting in severe economic loss. Meanwhile, traditional culturing method for detecting LAB are time-consuming for brewers. The present review introduces LAB as spoilage microbes in daily life, with focus on LAB in the brewing industry, targeting at the spoilage mechanism of LAB in brewing industry including the special metabolisms, the exist of the viable but nonculturable (VBNC) state and the hop resistance. At the same time, this review compares the traditional and novel rapid detection methods for these microorganisms which may provide innovative control and detection strategies for preventing alcoholic beverage spoilage, such as improvement of microbiological quality control using advanced culture media or different isothermal amplification methods.

Molecular Biologic Analysis of c-kit Gene in Salivary Gland Carcinoma (타액선암에서 c-kit 유전자에 대한 분자생물학적 연구)

  • Seo Kyu-Hwan;Jung Kwang-Yoon;Woo Jung-Soo;Baek Seung-Kuk;Choi Sung-Bae;Kim Sang-Hee;Kim In-Sun;Kwon Soon-Young
    • Korean Journal of Head & Neck Oncology
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    • v.19 no.2
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    • pp.121-126
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    • 2003
  • Objectives: The c-kit gene encodes a transmembrane receptor-type tyrosine kinase, which is known to have a significant role in the normal migration and development of germ cells and melanocytes. In the previous studies of c-kit gene, c-kit expressions showed only in adenoid cystic carcinomas, lymphoepithelioma-like carcinomas and myoepithelial carcinomas, but not in others and mutation was not found in any types of salivary carcinoma. We investigate the c-kit expression which may be useful to differentiating adenoid cystic carcinomas from others, and mutation of the gene which may not be exist nor the mechanism of c-kit activation in salivary carcinomas. Material and Methods: The archival tissue samples from 42 salivary carcinomas of major and minor salivary glands were studied for c-kit expression by immunohistochemistry and gene mutation by polymerase chain reaction amplification and single strand conformational polymorphism. Results: The c-kit expressions were noted in 22/24 adenoid cystic carcinomas, 7/9 mucoepidermoid carcinomas, 2/3 acinic cell carcinomas, 3/4 malignant mixed tumors, and one undifferentiated carcinoma. The mutation of c-kit gene was found in 3/24 adenoid cystic carcinomas, 3/8 mucoepidermoid carcinomas, one acinic cell carcinoma, and 2/4 malignant mixed tumors. Conclusion: c-kit protein overexpression is seen in a variety of salivary gland carcinomas, and the mutation of the gene may be the mechanism of c-kit activation in these neoplasms.

Generation and Growth of Long Ocean Waves along the West Coast of Korea in March 2007 (2007년 3월 한국 서해안에 발생한 해양장파의 형성과 성장과정)

  • Choi, Byoung-Ju;Park, Yong-Woo;Kwon, Kyung-Man
    • Ocean and Polar Research
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    • v.30 no.4
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    • pp.453-466
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    • 2008
  • In order to examine the generation mechanism of long ocean waves along the west coast of Korea and to understand the amplification process of the long ocean waves, sea level, atmospheric pressure and wind data observed every minute from 2007 March 29 to 2007 April 1 were analyzed and onedimensional numerical ocean model experiments were performed. An atmospheric pressure jump propagated southeastward from Backryungdo to Yeonggwang along the west coast of Korea with speed of $13{\sim}27\;m/s$ between 2007 March 30 23:00 and 2007 April 1 1:30. Average magnitude of pressure jump was 4.2 hPa. As a moving atmospheric jump propagated from north to south along the coast, long ocean waves were generated and the sea level abnormally rose or fell at Anheung, Kunsan, Wido and Yeonggwang. Average amplitude of sea level rise (or fall) was about 113.6 cm. In a one-dimensional numerical ocean model, nonlinear shallow water equations were numerically integrated and a moving atmospheric pressure jump with traveling speed of 24 m/s was used as an external force. While the atmospheric pressure jump travels over 60 m depth ocean, a long ocean wave is generated. Because the propagation speed of the atmospheric jump is almost equal to that of the long ocean wave, Proudman resonance occurs and the long ocean wave amplifies. As the atmospheric pressure jump moves into the coastal area shallower than 60 m, the speed of the long ocean wave decreases and Proudman resonance effect decreases. However, the amplitude of the long ocean wave increases and wave length becomes shorter because of shoaling effect. When the long ocean wave hits the land boundary, amplitude of the long ocean wave drastically amplifies due to reflection. Data analysis and numerical experiments suggest that the southeastward propagation of an atmospheric pressure jump over the shallow ocean, which is a necessary condition for Proudaman resonance, generated the long ocean waves along the west coast of Korea on 2007 March 31 and the ocean waves amplified due to shoaling effect in the coastal area and reflection at the shore.

Cloning and Characterization of a Cellulase Gene from a Plant Growth Promoting Rhizobacterium, Bacillus subtilis AH18 against Phytophthora Blight Disease in Red-Pepper (고추역병을 방제하는 PGPR균주 Bacillus subtilis AH18의 항진균성 Cellulase 유전자의 Cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Jung, Hee-Kyoung;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.311-317
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    • 2006
  • Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

Enhancement of TRAIL-Mediated Apoptosis by Genistein in Human Hepatocellular Carcinoma Hep3B Cells: Roles of p38 MAPK Signaling Pathway (인체간암세포에서 genistein의 TRAIL에 의한 apoptosis 유도 상승효과에서 미치는 p38 MAPK signaling pathway의 영향)

  • Jin, Cheng-Yun;Park, Cheol;Park, Sang-Eun;Hong, Sang-Hoon;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1549-1557
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    • 2011
  • Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can selectively induce apoptosis in many types of transformed cells; however, some human hepatocellular carcinoma cells are particularly resistant to the effects of TRAIL. Although genistein, a natural isoflavonoid phytoestrogen, has been shown to have pro-apoptotic activity against human cancer cell lines, little is known about the mechanism of genistein in terms of TRAIL-induced apoptosis. In the present study, it was investigated whether or not combined treatment with genistein and TRAIL synergistically induced apoptosis in Hep3B hepatocarcinoma cells. Results indicate that treatment with TRAIL in combination with nontoxic concentrations of genistein sensitized TRAIL-resistant Hep3B cells to TRAIL-induced apoptosis, which was associated with mitochondrial dysfunction. Further, the inhibition of p38 mitogen-activated protein kinase (MAPK) activation markedly decreased genistein and TRAIL-induced cell viability and apoptosis by enhanced truncation of Bid, increase of pro-apoptotic Bax, decrease of anti-apoptotic Bcl-2, and release of cytochrome c from mitochondria to cytoplasm. Activation of caspases and degradation of poly (ADP-ribose) polymerase induced by the combined treatment was also markedly increased by the inhibition of p38 MAPK, through the mitochondrial amplification step. In conclusion, our data suggest that genistein sensitizes TRAIL-induced-apoptosis via p38 MAPK-dependent pathway.

Expression of receptors of Vitamin D and cytokines in osteoclasts differentiated by M-CSF and ODF (Macrophage Colony-Stimulating Factor와 Osteoclast Differentiation Factor로 분화 유도된 생쥐 파골세포에서 Vitamin D 및 수종의 싸이토카인 수용체의 발현)

  • Seong, Soo-Mi;Um, Heung-Sik;Ko, Sung-Hee;Woo, Kyung-Mi;Chang, Beom-Seok
    • Journal of Periodontal and Implant Science
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    • v.32 no.4
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    • pp.865-873
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    • 2002
  • The primary cause of tooth loss after 30 years of age is periodontal disease. Destruction of alveolar bone by periodontal disease is done by bone resorbing activity of osteoclasts. Understanding differentiation and activation mechanism of osteoclasts is essential for controling periodontal disease. The purpose of this study is to identify the possible effects of Vitamin D and cytokines affecting osteoclasts and its precursor cells. Four to six week-old mice were killed and humerus, radius, tibia and femur were removed aseptically and washed two times with Hank's solution containing penicillin-streptomycin and then soft tissue were removed. Bone marrow cells were collected by 22 gauge needle. Cells were cultured in Hank's solution containing 1 mg/ml type II collagenase, 0.05% trypsin, 41mM EDTA. Supernatant solution was removed 5 times after 15 minutes of digestion with above mentioned enzyme solution, and remained bone particles were maintained in alpha-MEM for 15 minutes and $4^{\circ}C$ temperature. Bone particles were agitated for 1 minute and supernatant solution containing osteoclast precursor cells were filtrated with cell stainer. These separated osteoclast precursor cells were dispensed with 100-mm culture dish by $1{\times}10^7$ cells unit and cultured in ${\alpha}$- MEM containing 20 ng/ml recombinant human M-CSF, 30 ng/ml recombinant human soluble osteoclast differentiation factor and 10% fetal calf serum for 2 and 7 days. Total RNA of osteoclast precursor cells were extracted using RNeasy kit. One ${\mu}g$ of total RNA was reverse transcribed in $42^{\circ}C$ for 30 minutes using SuperScriptII reverse transcriptase. Expression of transcribed receptors of each hormone and cytokine were traced with 1 ${\mu}l$ of cDNA solution by PCR amplification. Vitamin D receptor WAS found in cells cultured for 7 days. TNF-${\alpha}$ receptor was found in cells cultured for 2 days and amount of receptors were increased by 7 days. IL-1 type I receptor was not found in cells cultured 2 and 7 days. But, IL-1 receptor type II was found in cells cultured for 2 days. TGF-${\alpha},{\beta}$type I receptor was found in cells cultured 2 and 7 days, and amount of receptors were increased by 7 days of culture. These results implies Vitamin D and cytokines can affect osteoclasts directly, and affecting period in differentiation cycle of osteoclasts is different by Vitamin D and cytokines.

Coat colour phenotype of Qingyu pig is associated with polymorphisms of melanocortin receptor 1 gene

  • Wu, Xiaoqian;Tan, Zhendong;Shen, Linyuan;Yang, Qiong;Cheng, Xiao;Liao, Kun;Bai, Lin;Shuai, Surong;Li, Mingzhou;Li, Xuewei;Zhang, Shunhua;Zhu, Li
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.7
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    • pp.938-943
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    • 2017
  • Objective: Qingyu pig, a Chinese indigenous pig breed, exhibits two types of coat colour phenotypes, including pure black and white with black spotting respectively. Melanocortin receptor 1 (MC1R) and agouti signaling protein (ASIP) are two widely reported pivotal genes that significantly affect the regulation of coat colour. The objectives of this study were to investigate whether the polymorphisms of these two genes are associated with coat colour and analyze the molecular mechanism of the coat colour separation in Qingyu pig. Methods: We studied the phenotype segregation and used polymerase chain reaction amplification and Sanger sequencing to investigate the polymorphism of MC1R and ASIP in 121 Qingyu pigs, consisting of 115 black and 6 white with black spotted pigs. Results: Coat colour of Qingyu pig is associated with the polymorphisms of MC1R but not ASIP. We only found 2 haplotypes, $E^{QY}$ and $E^{qy}$, based on the 13 observed mutations from MC1R gene. Among which, $E^{qy}$ presented a recessive inheritance mode in black spotted Qingyu pigs. Further analysis revealed a g.462-463CC insertion that caused a frameshift mutation and a premature stop codon, thus changed the first transmembrane domain completely and lost the remaining six transmembrane domains. Altogether, our results strongly support that the variety of Qingyu pig's coat colour is related to MC1R. Conclusion: Our findings indicated that black coat colour in Qingyu pig was dominant to white with black spotted phenotype and MC1R gene polymorphism was associated with coat colour separation in Qingyu pig.

First Detection of $bla_{IMP-1}$ in Clinical Isolate Multiresistant Acinetobacter baumannii from Korea

  • Jeong Seok-Hoon;Bae Il-Kwon;Sohn Seung-Ghyu;Park Kwang-Ok;An Young-Jun;Sung Kwang-Hoon;Jang Seon-Ju;Heo Myong-Jin;Yang Ki-Suk;Lee Sang-Hee
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1377-1383
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    • 2006
  • Among 46 Acinetobacter baumannii isolates collected in 2004, two imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Republic of Korea. Two carbapenemase-producing isolates were further investigated to determine the mechanism of resistance. These isolates were analyzed by antibiotic susceptibility testing, microbiological tests of carbapenemase activity, determination of pI, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. Two cases of infection by A. baumannii producing the IMP-1 ${\beta}$-lactamase were detected. The isolates were characterized by a modified cloverleaf synergy test and EDTA-disk synergy test. Isoelectric focusing of crude bacterial extracts revealed nitrocefin-positive bands with a pI value of 9.0. PCR amplification and characterization of the amplicons by direct sequencing indicated that the isolates carried a $bla_{IMP-l}$ determinant. The isolates were characterized by a multidrug resistance phenotype, including penicillins, extended-spectrum cephalosporins, carbapenems, and aminoglycosides. These results indicate that the observed imipenem resistance of two Korean A. baumannii isolates was due to the spread of an IMP-1-producing clone. Our microbiological test of carbapenemase activity is simple to screen class B metallo-${\beta}$-lactamase-producing clinical isolates to determine their clinical impact and to prevent further spread. This study shows that the $bla_{IMP-l}$ resistance determinant, which is emerging in Korea, may become an emerging therapeutic problem, since clinicians are advised not to use extended-spectrum cephalosporins, imipenem, and aminoglycosides. This observation emphasizes the importance of having effective control measures in Asian hospitals, such as early detection of colonized patients, isolation procedures, and a judicious use of antibiotics.

Rapid detection of Rifampicin- resistant M, tuberculosis by PCR-SSCP of rpoB gene (결핵균의 rpoB유전자 PCR-SSCP법에 의한 Rifampicin 내성의 신속 진단)

  • Shim, Tae Sun;Yoo, Chul-Gyu;Han, Sung Koo;Shim, Young-Soo;Kim, Young Whan
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.842-851
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    • 1996
  • Background : Rifampicin(RFP) is a key component of the antituberculous shon-course chemotherapy and the RFP-resistance is a marker of multi-drug resistant(MDR) M. tuberculosis. rpoB gene encodes the ${\beta}$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. Recent reports show that rpoB gene mutations are the cause of RFP resistance of M. tuberculosis and the main mechanism of rpoB gene mutation is point mutation. And PCR-SSCP is a rapid and easy method for detecting point mutations. So we performed PCR-SSCP of rpoB gene of M. tuberculosis and compared the result with traditional RFP sensitivity test. Method : The 27 RFP sensitive M. tuberculosis culture isolates and 25 RFP resistant isolates were evaluated. The RFP sensitivity test was done at the Korean Tuberculosis istitute. The DNA was extracted by bead beater method and was amplified with primers TR-8 and TR-9 in a 20ul PCR reaction containing 0.1ul(luCi) [${\alpha}-^{32}P$] - dCTP. After amplification, SSCP was done using non-denaturaring polyacrylamide gel electrophoresis. Then direct sequencing was done in cases of different eletrophoretic mobility compared with that of H37Rv. In 19 cases, we compared PCR-SSCP results with patient's clinical course and the results of traditional RFP sensitivity test. Results : 1) All 27 RFP sensitive M. tuberculosis isolates showed the same electrophoretic mobility compared with that of H37Rv. And all 25 RFP resistant M. tuberculosis isolates showed different electrophoretic mobility. 2) The mechanism of rpoB gene mutation of M. tuberculosis is mainly point mutation. 3) The PCR-SSCP results correlate well with traditional RFP sensitivity and patient's clinical response to antituberculous treatment. Conclusion: The PCR-SSCP of rpoB gene is a very sensitive and rapid mehod in detecting RFP- resistant M. tuberculosis.

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Differences of Illness Behavior in Depressive Patients According to the Presence of Somatization (우울증 환자에서 신체화 증상에 따른 질병행동의 차이)

  • Yoon, Chang-Young;Jang, Se-Heon;Jae, Young-Myo;Lee, Dae-Su;Choi, Jin-Hyuk
    • Korean Journal of Psychosomatic Medicine
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    • v.17 no.2
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    • pp.68-74
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    • 2009
  • Objectives : Illness behavior is defined as the persistence of an adaptive/maladaptive mode of perceiving, evaluating and responding to health status and symptoms according to the status. In a cognitive aspect, somatizing symptoms are regarded as being originated from distortions, including magnification and/or amplification, of perceiving, evaluating and responding to symptoms such as bodily sensations. That is somatization may be explained by maladaptive illness behavior. In this study, we tried to investigate differences of illness behavior in depressive patient according to the presence of somatization. Methods : We divided 45 patients who were diagnosed as depression with ICD-10 diagnostic criteria into two groups(somatizing and non-somatizing group) according to the somatization subscale of Korean Depression Scale and compared two groups in the differences of illness behavior using the Illness Behavior Questionnaire. Results : Somatizing group showed significantly higher scores than non-somatizing group on the disease affirmation subscale($6.79{\pm}2.08$ vs. 4.76, p=0.003) and the denial subscale($3.25{\pm}1.22$ vs. $2.10{\pm}1.41$, p=0.006). There were no significant differences between two groups on the general hypochondriasis subscale and the affective unstability subscale. In a logistic regression analysis, somatizing group also showed higher odds ratio (OR) scores on the disease affirmation subscale(OR=1.418, p=0.089) and the denial subscale(OR=1.880, p=0.083). Conclusion : The disease affirmation and denial may be a discriminative mechanism of somatization in depressive patients. These subscales of Illness Behavior Questionnare could be useful markers, and psychiatric illnesses with somatizing and depressive symptoms may be differentially diagnosed and be predicted through these subscales.

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