• Title/Summary/Keyword: molecular monitoring

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Characterization of Two Ricin Isoforms by Sodium Dodecyl Sulfate-Capillary Gel Electrophoresis and Capillary Isoelectric Focusing

  • Na, Dong-Hee;Park, Eun-Ji;Kim, Myung-Sun;Cho, Cheong-Kwan;Woo, Byung-Ho;Lee, Hye-Suk;Lee, Kang-Choon
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4253-4257
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    • 2011
  • Sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) and capillary isoelectric focusing (CIEF) were employed to characterize and compare ricin E purified from the small grain seeds of Ricinus communis with ricin D isoform. During the purification of ricin E using ion-exchange and size-exclusion chromatography, SDS-CGE was found to be useful for monitoring the efficiencies of purification steps. SDS-CGE showed that the molecular size of ricin E was not significantly different from that of ricin D, which was confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. CIEF was useful for discriminating ricin E from ricin D based on their isoelectric points (pI). The pI values of ricin E and D were 8.6-8.8 and 7.0-7.4, respectively. This study demonstrates the usefulness of SDS-CGE and CIEF for the characterization of ricin toxins.

Diversity of Nitrifying and Denitrifying Bacteria in SMMIAR Process (완전침지형 회전매체공정 내 질산화 및 탈질 관련 미생물의 군집 분포)

  • Quan, Zhe-Xue;Lim, Bong-Su;Kang, Ho;Yoon, Kyung-Yo;Yoon, Yeo-Gyo
    • Journal of Korean Society on Water Environment
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    • v.22 no.6
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    • pp.1014-1021
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    • 2006
  • SMMIAR (Submerged Moving Media Intermittent Aeration Reactor) Process is a very efficient system which remove ammonia to nitrogen gas in one reactor. In this study, we determined the diversity of ammonia oxidizing bacteria and denitrifying bacteria using specific PCR amplification and the clone library construction. An ammonia monooxygenase gene(amoA) was analyzed to investigate the diversity of nitrifiers. Most of amoA gene fragments (27/29, 93%) were same types and they are very similar (>99%) to the sequences of Nitrosomonas europaea and other clones isolated from anoxic ammonia oxidizing reactors. ANAMMOX related bacteria have not determined by specific PCR amplification. A nitrite reductase gene(nirK) was analyzed to investigate the diversity of denitrifying bacteria. About half (9/20, 45%) of denitrifiers were clustered with Rhodobacter and most of others were clustered with Mesorhizobium (6/20, 30%) and Rhizobium (3/20, 15%). All of these nirK gene clones were clustered in alpha-Proteobacteria and this result is coincide with other system which also operate nitrification and denitrification in one reactor. The molecular monitoring of the population of nitrifiers and denitrifiers would be helpful for the system stabilization and scale-up.

Sub-mm variability of a YSO (EC53) in Serpens main region : JCMT Transient survey

  • Yoo, Hyunju;Lee, Jeong-Eun;Johnstone, Doug;Mairs, Steve;Herczeg, Gregory
    • The Bulletin of The Korean Astronomical Society
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    • v.42 no.1
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    • pp.39.2-39.2
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    • 2017
  • Stars form through the gravitational collapse of molecular clouds. However, the rate at which a star gains most of its mass and the physics that drives the main phase of stellar growth is still unclear. The typical luminosity of observed protostars is smaller than what expected from the Shu's inside-out collapse model, which predicts a constant mass accretion rate. The episodic accretion model has been suggested as a solution of this luminosity problem. The JCMT Transient survey is a long term monitoring program using JCMT/SCUBA-2 to detect accretion variability of protostars in the eight nearby star-forming regions. Recently, we found a rise of the 850 micron flux at a clump in the Serpens main region at the rate of ~17% relative to the mean flux over previous observations. The submm clump is associated with a class I protostar, EC53, which has been reported as a binary system with a periodic variability. In this talk, we will provide a brief overview of the JCMT Transient Survey project, present the detection of the variable source, and discuss about follow-up observations.

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Hybrid Imaging in Oncology

  • Fatima, Nosheen;uz Zaman, Maseeh;Gnanasegaran, Gopinath;Zaman, Unaiza;Shahid, Wajeeha;Zaman, Areeba;Tahseen, Rabia
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.5599-5605
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    • 2015
  • In oncology various imaging modalities play a crucial role in diagnosis, staging, restaging, treatment monitoring and follow up of various cancers. Stand-alone morphological imaging like computerized tomography (CT) and magnetic resonance imaging (MRI) provide a high magnitude of anatomical details about the tumor but are relatively dumb about tumor physiology. Stand-alone functional imaging like positron emission tomography (PET) and single photon emission tomography (SPECT) are rich in functional information but provide little insight into tumor morphology. Introduction of first hybrid modality PET/CT is the one of the most successful stories of current century which has revolutionized patient care in oncology due to its high diagnostic accuracy. Spurred on by this success, more hybrid imaging modalities like SPECT/CT and PET/MR were introduced. It is the time to explore the potential applications of the existing hybrid modalities, developing and implementing standardized imaging protocols and train users in nuclear medicine and radiology. In this review we discuss three existing hybrid modalities with emphasis on their technical aspects and clinical applications in oncology.

A Continuous Spectrophotometric Assay for NADPH-cytochrome P450 Reductase Activity Using 1,1-Diphenyl-2-Picrylhydrazyl

  • Yim, Sung-Kun;Yun, Su-Jung;Yun, Chul-Ho
    • BMB Reports
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    • v.37 no.5
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    • pp.629-633
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    • 2004
  • NADPH-cytochrome P450 reductase (CPR) transfers electrons from NADPH to cytochrome P450, and catalyzes the one-electron reduction of many drugs and foreign compounds. Various forms of spectrophotometric titration have been performed to investigate the electron-accepting properties of CPR, particularly, to examine its ability to reduce cytochrome c and ferricyanide. In this study, the reduction of 1,1-diphenyl-2-picrylhydrazyl (DPPH) by CPR was assessed as a means of monitoring CPR activity. The principle advantage of DPPH is that its reduction can be assayed directly in the reaction medium by a continuous spectrophotometry. Thus, electrons released from NADPH by CPR were transferred to DPPH, and DPPH reduction was then followed spectrophotometrically by measuring $A_{520}$ reduction. Optimal assay concentrations of DPPH, CPR, potassium phosphate buffer, and NADPH were first established. DPPH reduction activity was found to depend upon the strength of the buffer used, which was optimal at 100 mM potassium phosphate and pH 7.6. The extinction coefficient of DPPH was $4.09\;mM^{-1}\;cm^{-1}$. DPPH reduction followed classical Michaelis-Menten kinetics ($K_m\;=\;28\;{\mu}M$, $K_{cat}\;=\;1690\;min^{-1}$). This method uses readily available materials, and has the additional advantages of being rapid and inexpensive.

Anticoagulant activities of oleanolic acid via inhibition of tissue factor expressions

  • Lee, Won-Hwa;Yang, Eun-Ju;Ku, Sae-Kwang;Song, Kyung-Sik;Bae, Jong-Sup
    • BMB Reports
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    • v.45 no.7
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    • pp.390-395
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    • 2012
  • Oleanolic acid (OA), a triterpenoid known for its anti-inflammatory and anti-cancer properties, is commonly present in several medicinal plants but its anticoagulant activities have not been studied. Here, the anticoagulant properties of OA were determined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), fibrin polymerization as well as cell-based thrombin and activated factor X (FXa) generation activities. Data showed OA prolonged aPTT and PT significantly and inhibited thrombin catalyzed fibrin polymerization. In addition, OA inhibited the activities of thrombin and FXa and inhibited the generation of thrombin or FXa in human endothelial cells. OA also inhibited TNF-${\alpha}$-induced tissue factor expression on human endothelial cells. In accordance with these anticoagulant activities, OA showed an anticoagulant effect in vivo. These results indicate that OA possesses antithrombotic activities and suggest that daily consumption of a herb containing OA may be preventing thrombosis in pathological states.

Biomarkers for the lung cancer diagnosis and their advances in proteomics

  • Sung, Hye-Jin;Cho, Je-Yoel
    • BMB Reports
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    • v.41 no.9
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    • pp.615-625
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    • 2008
  • Over a last decade, intense interest has been focused on biomarker discovery and their clinical uses. This interest is accelerated by the completion of human genome project and the progress of techniques in proteomics. Especially, cancer biomarker discovery is eminent in this field due to its anticipated critical role in early diagnosis, therapy guidance, and prognosis monitoring of cancers. Among cancers, lung cancer, one of the top three major cancers, is the one showing the highest mortality because of failure in early diagnosis. Numerous potential DNA biomarkers such as hypermethylations of the promoters and mutations in K-ras, p53, and protein biomarkers; carcinoembryonic antigen (CEA), CYFRA21-1, plasma kallikrein B1 (KLKB1), Neuron-specific enolase, etc. have been discovered as lung cancer biomarkers. Despite extensive studies thus far, few are turned out to be useful in clinic. Even those used in clinic do not show enough sensitivity, specificity and reproducibility for general use. This review describes what the cancer biomarkers are for, various types of lung cancer biomarkers discovered at present and predicted future advance in lung cancer biomarker discovery with proteomics technology.

Feasibility of simultaneous measurement of cytosolic calcium and hydrogen peroxide in vascular smooth muscle cells

  • Chang, Kyung-Hwa;Park, Jung-Min;Lee, Moo-Yeol
    • BMB Reports
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    • v.46 no.12
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    • pp.600-605
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    • 2013
  • Interplay between calcium ions ($Ca^{2+}$) and reactive oxygen species (ROS) delicately controls diverse pathophysiological functions of vascular smooth muscle cells (VSMCs). However, details of the $Ca^{2+}$ and ROS signaling network have been hindered by the absence of a method for dual measurement of $Ca^{2+}$ and ROS. Here, a real-time monitoring system for $Ca^{2+}$ and ROS was established using a genetically encoded hydrogen peroxide indicator, HyPer, and a ratiometric $Ca^{2+}$ indicator, fura-2. For the simultaneous detection of fura-2 and HyPer signals, 540 nm emission filter and 500 nm~ dichroic beamsplitter were combined with conventional exciters. The wide excitation spectrum of HyPer resulted in marginal cross-contamination with fura-2 signal. However, physiological $Ca^{2+}$ transient and hydrogen peroxide were practically measurable in HyPer-expressing, fura-2-loaded VSMCs. Indeed, distinct $Ca^{2+}$ and ROS signals could be successfully detected in serotonin-stimulated VSMCs. The system established in this study is applicable to studies of crosstalk between $Ca^{2+}$ and ROS.

Detection of DNA Hybridization Characteristics Using Electrochemical methods (전기화학법을 이용한 DNA Hybridization 특성 검출)

  • Kim, Do-Kyun;Chang, Jeong-Soo;Kwon, Young-Soo
    • Proceedings of the KIEE Conference
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    • 2002.07c
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    • pp.1569-1571
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    • 2002
  • The determination of DNA hybridization can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, The determination of hybridization is very important for the improvement of DNA detection system. In this study, we report the characterization of the DNA hybridization by the electricalchemical methods. A new electrochemical biosensor is described for voltammetric detection of gene sequence related to probe oligonucleotide of bacterium Escherichia coli O157:H7. The biosensor involves the immobilization of a 18-mer probe oligonucleotide, which is complemetary to a specific gene sequence related to Escherichia coli O157:H7 on a gold electrode through specific adsorption. The probe oligonucleotide was used to determine the amount of target oligonucleotide in solution using mitoxantrone(MTX) as the electrochemical indicators. The cathodic peak currents $(I_{peak})$ of MTX were linearly related to the concentration of the target oligonucleotide sequence in the range $1[{\mu}M]{\sim}0.1[nM]$. The detection limit of this approach was 0.01[nM]. In addition, these indicators were capable of selectivity discriminating against various mismatching condition.

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Transcriptional Profile and Cellular Effects on Time Course & Doses Treatment of Methylmercury using Human cDNA Microarray System

  • Kim, Youn-Jung;Yun, Hye-Jung;Kim, Eun-Young;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.176-176
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    • 2003
  • Methylmercury is known to have devastating effects on the mammalian nervous system. When human neuroblastoma SH-SY5Y cells were treated with methylmercury at sublethal concentrations (6.25 uM), up-regulated genes (39) & down-regulated genes (19) were identified by human 8k cDNA microarray. These genes are related with microtubule process, signal transduction pathway and cell death (apoptosis), Apoptosis-associated genes, HSP70, CDK inhibitor 1, FOS-like antigen were up-regulated and microtubule related genes like villin and dynein down-regultaed. To confirm the presence of apoptosis in cultured SH-SY5Y cells treated 6.25 and 1 uM methylmercury, we applied Annexin V-FITC assay followed by flow cytometric measurements after 6 and 24h. Studies on transcriptional and molecular effect by methylmercury may provide an insight into the neurotoxic effects of methylmercury in human neuronal cells and a possibility to develop more efficient and exact monitoring system of heavy metals as ubiquitous environmental pollutants.

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