• Title/Summary/Keyword: Bip

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Detection of Clostridium difficile by Loop-Mediated Isothermal Amplification (등온증폭법을 이용한 Clostridium difficile 검출)

  • In, Ye-Won;Ha, Su-Jeong;Yang, Seung-Kuk;Oh, Se-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.9
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    • pp.1326-1330
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    • 2012
  • This study was conducted to develop a loop-mediated isothermal amplification (LAMP) method for the detection of Clostridium difficile. The tested target gene was 16S ribosomal RNA. Five different LAMP primer sets were designed, and LAMP was performed. All primer sets targeting the 16S rRNA gene (BIP, FIP, B3, F3, LF, PF) were determined as positive in tcdA-positive, tcdB-postive ($A^+B^+$) and tcdA-negative, tcdB-negative ($A^-B^-$) Clostridium difficile strains. As the LAMP reaction took less than 80 min and did not require expensive machine such as thermocycler, it can be used as a rapid and simple detection method for foodborne pathogens.

Induction of HaCaT Cell Apoptosis by Sodium Nitroprusside (Sodium Nitroprusside로 유발한 HaCaT Cell의 Apoptosis)

  • Park, Yuri;Moon, Cheol;Kim, Sa-Hyun;Lee, Pyeongjae
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.3
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    • pp.112-116
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    • 2015
  • Nitric Oxide (NO) has been known to play important physiological and pathological roles. In this study, Sodium nitroprusside (SNP), NO donor, induced the apoptosis of HaCaT cell, human spontaneous immortal keratinocyte, which was investigated through DAPI staining and cleavage of PARP and caspase-3 protein. However, the expression level of Bip and CHOP, involved in ER stress, was not significantly changed as compared to the control cell group. Recent studies have showed that SIRT1, $NAD^+$-dependent deacetylase, is the key protein that controls cell survival and death. SNP treatment suppressed the SIRT1 gene expression, which indicated that apoptosis induced by SNP could be implicated in SIRT1 down-regulation.

Oxymatrine Causes Hepatotoxicity by Promoting the Phosphorylation of JNK and Induction of Endoplasmic Reticulum Stress Mediated by ROS in LO2 Cells

  • Gu, Li-li;Shen, Zhe-lun;Li, Yang-Lei;Bao, Yi-Qi;Lu, Hong
    • Molecules and Cells
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    • v.41 no.5
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    • pp.401-412
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    • 2018
  • Oxymatrine (OMT) often used in treatment for chronic hepatitis B virus infection in clinic. However, OMT-induced liver injury has been reported. In this study, we aim to investigate the possible mechanism of OMT-induced hepatotoxicity in human normal liver cells (L02). Exposed cells to OMT, the cell viability was decreased and apoptosis rate increased, the intracellular markers of oxidative stress were changed. Simultaneously, OMT altered apoptotic related proteins levels, including Bcl-2, Bax and pro-caspase-8/-9/-3. In addition, OMT enhanced the protein levels of endoplasmic reticulum (ER) stress makers (GRP78/Bip, CHOP, and cleaved-Caspase-4) and phosphorylation of c-Jun N-terminal kinase (p-JNK), as well as the mRNA levels of GRP78/Bip, CHOP, caspase-4, and ER stress sensors (IREI, ATF6, and PERK). Pre-treatment with Z-VAD-fmk, JNK inhibitor SP600125 and N-acetyl-l-cysteine (NAC), a ROS scavenger, partly improved the survival rates and restored OMT-induced cellular damage, and reduced caspase-3 cleavage. SP600125 or NAC reduced OMT-induced p-JNK and NAC significantly lowered caspase-4. Furthermore, 4-PBA, the ER stress inhibitor, weakened inhibitory effect of OMT on cells, on the contrary, TM worsen. 4-PBA also reduced the levels of p-JNK and cleaved-caspase-3 proteins. Therefore, OMT-induced injury in L02 cells was related to ROS mediated p-JNK and ER stress induction. Antioxidant, by inhibition of p-JNK or ER stress, may be a feasible method to alleviate OMT-induced liver injury.

Aerobic Exercise Ameliorated High Fat Diet-induced Endoplasmic Reticulum Stress More Than Polyphenol Supplementation in Skeletal Muscle of Obese Mice (비만에 의해 유도된 근형질세망 스트레스에서 유산소 운동에 의한 감소효과)

  • Park, Jong-Gu;Kim, Yong-An;Park, Hee-Geun;Lee, Wang-Lok
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1186-1192
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    • 2018
  • The purpose of this study was to compare the effects of either aerobic exercise or polyphenols supplementation on mRNA expression of endoplasmic reticulum stress in skeletal muscle of high fat diet-induced obese mice. In the study, mice were divided into five groups: (1) NC (normal diet for 16 weeks as a control, n=10), (2) HC (high fat diet for 16 weeks as a control, n=10), (3) H-Re (high fat diet with resveratrol 25 mg/kg supplementation for 16 weeks, n=10), (4) H-Ch (high fat diet with chrysin 50 mg/kg supplementation for 16 weeks, n=10), and (5) HE (high fat diet with aerobic exercise for 16 weeks, n=10). Aerobic exercise was performed on a treadmill for 40~60 min/day at 10~14 m/min, 0% grade, four days/week for 16 weeks. Endoplasmic reticulum stress related genes were measured by real-time polymerase chain reaction. ATF6, PERK, $IRE1{\alpha}$, and BIP/GRP78 mRNA were significantly decreased in HE compared with those in HC (p<0.05). Also, ATF6, $IRE1{\alpha}$, and BIP/GRP78 mRNA were significantly decreased in H-Re compared with those in HC (p<0.05). ATF6 mRNA was significantly decreased in H-Ch compared with that in HC (p<0.05). These findings suggest that aerobic exercise, resveratrol, and chrysin supplementation changed ER stress markers. However, aerobic exercise was most effective on ameliorating the high fat diet induced ER stress markers. Thus, it seems that aerobic exercise might have a more positive effect on skeletal muscle endoplasmic reticulum stress compared with polyphenol supplementation in high fat diet-induced obese mice.

Effects of High Stocking Density on the Expressions of Stress and Lipid Metabolism Associated Genes in the Liver of Chicken (닭의 고밀도 사양체계가 스트레스 및 지방대사 연관 유전자 발현에 미치는 영향)

  • An, Young Sook;Park, Jeong Geun;Jang, In Surk;Sohn, Sea Hwan;Moon, Yang Soo
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1672-1679
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    • 2012
  • The effect of high stocking density (HSD) on the expression of stress and lipid metabolism associated genes in the liver of broiler chickens was examined by chicken genome array analysis. The chickens in a control group were randomly assigned to a $495cm^2/bird$ stocking density, whereas the chickens in a HSD group were arranged in a $245cm^2/bird$ stocking density with feeding ad libitum for 35 days. The chickens assigned to the HSD group had a significantly lower body weight, weight gain, and feed intake compared with those of the control group (p<0.05). The mortality of chickens was higher in the HSD group than in the control group. The microarray analysis indicated up-regulation of stress associated genes such as HMGCR, $HSP90{\alpha}$, HSPA5 (GRP78/Bip), DNAJC3 and ATF4, and down-regulation of interferon-${\gamma}$ and PDCD4 genes. The endoplasmic reticulum stress associated genes, HSPA5 (GRP78/Bip), DNAJC3 and ATF4, were highly expressed in the HSD group. The genes, ACSL5, TMEM195 and ELOVL6, involved in fatty acid synthesis, were elevated in the HSD group. The genes, ACAA1, ACOX1, EHHADH, LOC423347 and CPT1A, related to fatty acid oxidation, were also activated in the HSD group. These results suggest that a HSD rearing system stimulates the genes associated with fatty acid synthesis as well as fatty acid oxidation in the liver of broiler chickens.

Detection of Salmonella typhi by Loop-mediated Isothermal Amplification Assay

  • Jo, Yoon-Kyung;Lee, Chang-Yeoul
    • Biomedical Science Letters
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    • v.14 no.2
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    • pp.115-118
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    • 2008
  • Salmonella typhi is frequent causes of foodborne illness and its detection is important for monitoring disease progression. In this study, by using general PCR and novel LAMP (Loop Mediated Isothermal Amplification) assay, we evaluated the usefulness of LAMP assay for detection of Salmonella typhi. In this LAMP assay, forward inner primer (FIP) and back inner primer (BIP) was specially designed for recognizing target invA gene. Target DNA was amplified and visualized as ladder-like pattern of bands on agarose gel within 60 min under isothermal conditions at $65^{\circ}C$. When the sensitivity and reproducibility of LAMP were compared to general PCR, there was no difference of reproducibility but sensitivity of LAMP assay was more efficient than PCR (the detection limit of LAMP assay was 30 fg, while the PCR assay was 3 pg). These results indicate that the LAMP assay is a potential and valuable means for detection of Salmonella typhi, especially for its rapidity, simplicity and low cost.

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시추공 유속측정기(Borehole Flowmeter)를 이용한 암반의 구간별 수리전도도 산정

  • 구민호;차장환;이주형;박창희;정복선
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2001.04a
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    • pp.93-97
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    • 2001
  • 시추공의 구간별 수리전도를 추정하는 방법으로는 수압시험(packer test)이 많이 이용되는데 최근에는 유속측정기(flowmeter)를 이용한 시추공 검층법이 개발되어 활발한 연구가 진행 중이다. 본 연구에서는 열원(heat-pulse) 공급 방식의 유속측정기를 이용하여 공주대학 교내에 설치된 시추공에서 자연 유속(ambient flow) 및 양수 유발 유속(pump-induced flow)을 측정하였으며, 자료를 분석하여 수리전도도의 수직적인 분포를 산정하였다. 분석 결과는 수압시험에 의해 산정된 수리전도도의 분포와 잘 일치하였으며, BIPS에 의해 촬영된 시추공 영상 자료와 비교함으로써 지하수 유동과 관련된 투수성 단열(conductive fracture) 들의 수직적인 위치를 정확하게 파악할 수 있었다. 분석 결과는 암반 대수층 내에 발달된 단열망(fracture network)에 대한 3차원적인 정보를 제공할 수 있으며, 이는 효과적인 지하수 모니터링, 모델링, 및 정화 설계(remedial design)에 필요한 기초 자료로 활용될 수 있을 것으로 기대된다.

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Energy Efficient MST based Broadcast with Hitch-hiking in Wireless Ad Hoc Network (Hitch-hiking 기법을 이용한 MST 기반 무선 애드 혹 네트워크 브로드캐스트 알고리즘)

  • Jeon, Sang-Hyun;Chang, Hyeong-Soo;Cho, Joon-Ho
    • Proceedings of the Korean Information Science Society Conference
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    • 2006.10d
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    • pp.211-216
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    • 2006
  • 본 논문에서는 정적인(static) 무선 애드 혹 네트워크(wireless ad hoc network)상에서 동작하는 에너지 효율적인 broadcast알고리즘인 "MST based Broadcast with Hitch-hiking(MBH)"알고리즘을 제안한다. MBH는 BIP[1]에 Hitch-hiking기법[2]을 적용시킨 BHH(Broadcast by Hitch-Hiking)[3]에 비해 적은 시간 복잡도를 가지면서 더 적은 에너지를 소모해 broadcast를 수행할 수 있음을 실험을 통해 확인할 수 있었다. 또한 분산(distributed) MBH는 1-hop 이웃정보만을 이용해 broadcast 트리를 구성하기 때문에 높은 확장성(scalability)을 가지며, 빠르고 효율적으로 에너지 효율적인 broadcast 트리를 구성할 수 있다. 또한 분산 MBH는 이동성을 고려한 동적 네트워크의 잦은 위상변화에 빠르게 적응할 수 있도록 쉽게 확장할 수 있다.

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A Loop-mediated Isothermal Amplification Method for White-backed Planthopper-specific Detection (고리매개등온증폭법(LAMP)을 이용한 흰등멸구 특이 판별법)

  • Seo, Bo Yoon;Park, Chang Gyu;Jung, Jin Kyo;Cho, Jumrae;Lee, Gwan-Seok;Kim, Kwang-Ho
    • Korean journal of applied entomology
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    • v.57 no.4
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    • pp.393-399
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    • 2018
  • A loop-mediated isothermal amplification (LAMP) primer set (WBPH-65) was designed for the species-specific detection of white-backed planthopper (WBPH) Sogatella furcifera based on the full-length sequence of the internal transcribed spacer 2 (ITS2) (KC417469.1). The WBPH-65 primer set consists of six primers (total 165 bp), F3 (18 bp), B3 (18 bp), FIP (43 bp), BIP (40 bp), LF (21 bp), and LB (25 bp). After the LAMP reaction of three rice planthoppers, S. furcifera, Nilaparvata lugens, and Laodelphax striatellus, with the WBPH-65 primer set for 60 min at $65^{\circ}C$, the LAMP products were observed in the genomic DNA of S. furcifera only. According to the DNA amount of S. furcifera and incubation duration at $65^{\circ}C$, the difference of fluorescence relative to the negative control (0 ng) was clearly observed in a 40-min incubation with 10 and 100 ng or in case of 60-min incubation with 0.01, 0.1, 1, 10, and 100 ng. There was little difference in fluorescence between the negative control and all the other DNAs tested in 20- and 30-min incubations. On the other hand, the WBPH-65 primer set without LF and LB primers showed little amplification in the genomic DNAs of the three rice planthoppers, S. furcifera, N. lugens, and L. striatellus in a 60-min incubation. These results suggest that all six primers (F3, B3, FIP, BIP, LF, and BF) are necessary for the WBPH-65 primer set to detect S. furcifera within a 60-min incubation, and is able to discriminate S. furcifera from at least N. lugens and L. striatellus.

Detection of Mycobacterium Tuberculosis by Loop-Mediated Isothermal Amplification Assay (등온 증폭법을 이용한 결핵균의 빠른 검출 시스템 개발)

  • Ahn, Young-Chang;Nam, Youn-Hyoung;Park, Su-Min;Cho, Min-Ho;Seo, Jae-Won;Yoon, Il-Kyu;Park, Yong-Hyun;Jang, Won-Cheoul
    • Journal of the Korean Chemical Society
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    • v.52 no.3
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    • pp.273-280
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    • 2008
  • Mycobacterium tuberculosis (MTB) remains a major worldwide public health problem. In recent years, the incidence of MTB has been rising. Rapid and reliable diagnosis of Mycobacterium tuberculosis is essential to initiate correct treatment, avoid severe complications, and prevent transmission. LAMP was used to develop a rapid and sensitive laboratory diagnostic system for the MTB. In this research, the loop-mediated isothermal amplification method (LAMP) that amplifies DNA with high specificity and rapidity at an isothermal condition was evaluated for rapid detection of MTB. Undiluted DNA (2.10 × 106 copy/mL), 10-1, 10-2, 10-3, 10-4, 10-5 and 10-6 (copy/mL) of MTB DNA were amplified by PCR and LAMP to determine the sensitivity of the assay. At results, the LAMP assay reported here has the advantages of rapid amplification, high sensitivity, and high specificity and will be useful for rapid and reliable clinical diagnosis of MTB in hospital clinical laboratory.