• Title/Summary/Keyword: Molecular techniques

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Analysis of Dominant Microorganisms of Bulking Sludge at Low Dissolved Oxygen Concentration using 16S rRNA Sequences (16S rRNA 염기서열을 이용한 낮은 용존산소농도에서 발생한 벌킹슬러지의 우점종 분석)

  • Kim, Yun-Jung;Park, Eun-Hye;Kim, Gyu-Dong;Nam, Kyoungphile;Chung, Tai Hak
    • Journal of Korean Society on Water Environment
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    • v.20 no.5
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    • pp.506-511
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    • 2004
  • Maintaining dissolved oxygen (DO) at sufficiently low concentration in the aeration tank at a wastewater treatment plant (WWTP) is essential for reduction of the costs of operation and maintenance. On the other hand, the low DO level may result in adverse effect on the integrity of the activated sludge, A typical and disastrous outcome frequently experienced is the outgrowth of filamentous microorganisms, which is called as filamentous bulking, In addition to the traditional methods such as sludge settleability and microscopic observation of the culture, molecular techniques including polymerase chain reaction (PCR) amplification followed by 16S rRNA sequencing were applied to identify filamentous bacteria present in bulking sludge under a condition of low DO concentration, Two morphologically distinct groups, presumably consisting of Sphaerofilus nafans, and Eikelboom Type 1701 or Type 1851, were identified through microscopic observation. They were further confirmed by subsequent 16S rRNA sequencing. Dominant filamentous bacteria identified by the molecular techniques were consisted of three major groups. Sequences of partial 16S rRNA cloned showed that the filamentous bulking organisms were closely related to Eikelboom Type 021N and Eikelboom Type 1701, and Sphaerotilus natans, respectively. Molecular methods were found to possess a strong potential of direct examination of the microbial community of an activated sludge system.

Alteration of voltage-dependent activation by a single point mutation of a putative nucleotide-binding site in large-conductance $Ca^{2+}$-activated $K^+$ channel

  • Kim, Hyun-Ju;Lim, Hyun-Ho;Park, Chul-Seung
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.44-44
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    • 2003
  • $BK_{Ca}$ channels were suggested to contain one or more domains of the ‘regulator of K+ conductance’(RCK) in their cytosolic carboxyl termini (Jiang et al.2001). It was also shown that the RCK domain in mammalian $BK_{Ca}$ channels might sense the intracellular $Ca^{2+}$ with a low affinity (Xia et al. 2002). We aligned the amino acid sequence of the $\alpha$-subunit of rat $BK_{Ca}$ channels (rSlo) with known RCK domains and identified a second region exhibiting about 50% homology. This putative domain, RCK2, contains the characteristic amino acids conserved in other RCK domains. We wondered whether this second domain is involved in the domain-domain interaction and the gating response to intracellular $Ca^{2+}$ for rSlo channel, as revealed in the structure of RCK domain of E. coli channel (Jiang et al.2001). In order to examine the possibility, site-directed mutations were introduced into the RCK2 domain of rSlo channel and the mutant channels were expressed in Xenopus oocytes for functional studies. One of such mutation, G772D, in the putative nucleotide-binding domain resulted in the enhanced $Ca^{2+}$ sensitivity and the channel gating of rSlo channel. These results suggest that this region of $BK_{Ca}$ channels is important for the channel gating and may form an independent domain in the cytosolic region of $BK_{Ca}$ channels. In order to obtain the mechanistic insights of these results, G772 residue was randomly mutagenized by site-directed mutagenesis and total 17 different mutant channels were constructed. We are currently investigating these mutant channels by electrophysiological techniques.ical techniques.

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The Role of Functional Feed Additives in Modulating Intestinal Health and Integrity

  • Kocher, Andreas
    • Korean Journal of Poultry Science
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    • v.39 no.1
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    • pp.33-37
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    • 2012
  • One of the biggest challenges for the animal feed industry in the coming years will be to meet the growing demand in animal protein in light of increased cost of feed ingredient as well as tougher restrictions on the use of antimicrobial growth promoters imposed by consumers and governments. A key focus area will be to maximise feed efficiency and minimise nutrient waste. It has been widely acknowledged that the composition of the intestinal microflora is closely related to intestinal health and performance of animals. Advanced microbial techniques have shown a close relationship between bacterial communities and their ability to modulate nutrient absorption and processing. In addition it has been recognised that modulating the immune response has significant impact on overall health as well as overall nutrient demand. Molecular techniques are a useful tool to gain an understanding of the impact of dietary interventions including the use of functional feed additives on specific changes in microbial communities or the immune system. Most these techniques however focus on the evaluation of large changes in bacterial compositions and often underestimate or neglect to recognise small changes in microbial diversity or behaviour changes without any measurable immune response. The key to understanding the relationship between specific nutritional intervention and the impact on health and performance lies in a deeper understanding of the impact of these nutrients on the expression of specific genes or specific metabolic pathways. The development of molecular tools as a result of developments in the field of Nutrigenomics has enabled researchers to study the effects of specific nutrients on the whole genome or in other words, the effect of thousands of genes simultaneously, and has opened a completely different avenue for nutritional research.

Application of extraction chromatographic techniques for separation and purification of emerging radiometals 44/47Sc and 64/67Cu

  • Vyas, Chirag K.;Park, Jeong Hoon;Yang, Seung Dae
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.2 no.2
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    • pp.84-95
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    • 2016
  • Considerably increasing interest in using the theranostic isotopes/ isotope pairs of radiometals like $^{44/47}Sc$ and $^{64/67}Cu$ for diagnosis and/or therapeutic applications in the nuclear medicine procedures necessitates its reliable production and supply. Separation and purification of no-carrier-added (NCA) isotopes from macro quantitates of the irradiated target matrix along with other impurities is a cardinal procedure amongst several other steps involved in its production. Multitudinous methods including but not limited to liquid-liquid (solvent) extraction, extraction chromatography (EXC), ion exchange, electrodeposition and sublimation are routinely applied either solitarily or in combination for the separation and purification of radioisotopes depending on their production routes, radioisotope of interest and impurities involved. However, application of EXC though has shown promises towards the numerous separation techniques have not received much attention as far as its application prospects in the field of nuclear medicine are concerned. Advances in the recent past for application of the EXC resins in separation and purification of the several medically important radioisotopes at ultra-high purity have shown promising behavior with respect to their operation simplicity, acidic and radiolytic stability, separation efficiencies and speedy procedures with the enhanced and excellent extraction abilities. In this mini review we will be talking about the recent developments in the application and the use of EXC techniques for the separation and purification of $^{44/47}Sc$ and $^{64/67}Cu$ for medical applications. Furthermore, we will also discuss the scientific and practical aspects of EXC in the view of separation of the NCA trace amount of radionuclides.

Advanced Analytical Techniques for Dissolved Organic Matter and Their Applications in Natural and Engineered Water Treatment Systems (최근 용존 유기물 분석 기법 및 자연환경과 수 처리 시스템 내 활용방안)

  • Lee, Yun Kyung;Hur, Jin
    • Journal of Korean Society on Water Environment
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    • v.38 no.1
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    • pp.31-42
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    • 2022
  • Dissolved organic matter (DOM), which changes according to various factors, is ubiquitously present from natural environments to engineered treatment systems. Only limited information is available regarding the environmental functions of DOM after bulk analyses are only applied for characterization. In this paper, latest DOM analytical techniques are briefly introduced, which include fluorescence excitation-emission matrix with parallel factor analysis (EEM-PARAFAC), size-exclusion chromatography with an organic carbon detector (SEC-OCD), carbon/nitrogen stable-isotope ratio, and Fourier transform-ion cyclotron resonance-mass spectroscopy (FT-ICR-MS). Recent examples of using advanced analyses to interpret the phenomena associated with DOM occurring in natural and engineered systems are presented here. Through EEM-PARAFAC, different components like protein-like, fulvic-like, and humic-like can be identified and tracked individually through the investigated systems. SEC-OCD allows researchers to quantify different size fractions. FT-ICR-MS provides thousands of molecular formulas present in bulk DOM samples. Lastly, carbon/nitrogen stable-isotope ratio offers reasonable tools for tracking the sources in environments. We also discuss the advantages and weakness of the above-mentioned characterizing tools. Specifically, they focus on single environmental factors (different sourced-DOM and interaction of sediment-pore water) or simple changes after individual treatment processes. Through collaboration with the advanced techniques later, they help the researchers to better understand environmental behaviors in aquatic systems and serve as essential tools for addressing various pending problems associated with DOM.

Promoting and improving three Rs practice: the Korean guidelines

  • Choe, Byung In;Lee, Gwi Hyang
    • BMB Reports
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    • v.50 no.12
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    • pp.610-614
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    • 2017
  • Scientists planning research that involves the use of animals are required to examine the possibilities for replacement, reduction, or refinement (the Three Rs), and their protocol must be reviewed by the Institutional Animal Care and Use Committee. Good scientific practice involving appropriate search techniques on the available Three Rs resources is essential for both ethical and scientific reasons. Appropriate experimental design and statistical analysis techniques are particularly necessary for research involving the use of animals, because this can improve animal welfare and scientific outcomes, as well as saving animal lives. There are a number of resources to help researchers improve their search techniques, experimental design strategies, and their reporting of research involving the use of animals. However, there is little specific information or resources on the Three Rs alternatives that is readily available in the Korean language. This paper outlines the common errors made by submitting researchers that have been repeatedly observed during the ethical review of experimental protocols over the last ten years, and provides information on the Korean resources available to promote good scientific practice. This could help to bridge the gap between Korean scientists and animal welfare advocates assisting scientists to improve ethical considerations and conduct responsible research.

Development of a general purpose molecular simulation system from microscopic to mesoscopic scales (미시영역에서 중간역역까지 적용 가능한 범용 분자 시뮬레이션 시스템의 개발)

  • Oh, Kwang-Jin
    • The KIPS Transactions:PartD
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    • v.12D no.6 s.102
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    • pp.921-930
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    • 2005
  • In this paper, a general purpose molecular simulation system which has been developed by the author, are described. One of the most advantageous features is that the molecular simulation system can handle a coarse-grained model as well as an all-atom mode. Therefore, we can simulate mesoscopic phenomena as well as microscopic phenomena with the help of Langevin dynamics simulation and dissipative particle dynamics simulation techniques. Thus we could study anesthesia, protein folding, biopolymer flow in microchannel with single framework, which spans from microscopic to mesoscopic scales. We expect that we can also simulate many other bio/nano systems of technological importance which are not feasible by means of molecular dynamics simulation technique. Finally, performance data are shown and a bottleneck is identified for future optimization.

Herbal Remedies for Combating Irradiation: a Green Anti-irradiation Approach

  • Lachumy, Subramanion Jothy;Oon, Chern Ein;Deivanai, Subramanian;Saravanan, Dharmaraj;Vijayarathna, Soundararajan;Choong, Yee Siew;Yeng, Chen;Latha, Lachimanan Yoga;Sasidharan, Sreenivasan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5553-5565
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    • 2013
  • Plants play important roles in human life not only as suppliers of oxygen but also as a fundamental resource to sustain the human race on this earthly plane. Plants also play a major role in our nutrition by converting energy from the sun during photosynthesis. In addition, plants have been used extensively in traditional medicine since time immemorial. Information in the biomedical literature has indicated that many natural herbs have been investigated for their efficacy against lethal irradiation. Pharmacological studies by various groups of investigators have shown that natural herbs possess significant radioprotective activity. In view of the immense medicinal importance of natural product based radioprotective agents, this review aims at compiling all currently available information on radioprotective agents from medicinal plants and herbs, especially the evaluation methods and mechanisms of action. In this review we particularly emphasize on ethnomedicinal uses, botany, phytochemistry, mechanisms of action and toxicology. We also describe modern techniques for evaluating herbal samples as radioprotective agents. The usage of herbal remedies for combating lethal irradiation is a green anti-irradiation approach for the betterment of human beings without high cost, side effects and toxicity.

Purification and Characterization of Membrane-Bound Phosphatidylinositol 4-Kinase from Mouse Brain

  • Lee, Sang-Min;Son, Hyeog-Gin;Lee, Young-Seek;Lee, Kang-Suk;Rhee, Sue-Goo;Cho, Key-Seung
    • BMB Reports
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    • v.29 no.6
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    • pp.555-563
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    • 1996
  • A membrane-bound phosphatidylinositol 4-kinase (PI 4-kinase) was separated in a sucrose gradient and solubilized with 1% Triton X-100 from mouse brain. The enzyme was purified 2,952-fold by various chromatographic techniques including DEAE-cellulose, PI-Sepharose and Sephacryl S-200 gel filtration. The molecular weight of PI 4-kinase was approximately 76 kDa by gel filtration and 70.8 kDa by SDS-polyacrylamide gel electrophoresis. The purified enzyme exhibited specific activity of 11.2 nmol/min/mg protein and pi value of 4.7. Kinetic analysis of the PI 4-kinase indicated apparent $K_m$, values of 190 ${\mu}M$ and 120 ${\mu}M$ for phosphatidylinositol and ATP, respectively. The maximal activity of this purified enzyme was observed at pH 7.4 at an incubation temperature of $37^{\circ}C$. The enzyme activity was significantly activated by $Mg^{2+}$, $Mn^{2+}$ and $Fe^{2+}$, and inhibited severely by $Ca^{2+}$. PI 4-kinase was proved to be pure in its immunoblot test by polyclonal antibody prepared from immunized rabbit sera. By this test, we were able to detect the existence of the same type of PI 4-kinase from other mouse organ tissues, such as liver, heart, kidney and spleen. Furthermore, similar immunoblot analysis with the same antisera recognized the different epitopes of PI 4-kinase proteins from various organs of rabbit, chinese hamster and rat.

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Recent Advances in Cancer Diagnosis: On an Overview of Diagnostic Cytopathologic Modalities and Ancillary Techniques (세포병리학적 기초에 의한 암진단의 발전: 진단방법과 보조기법)

  • Kim, Ki-Tai;Ham, Eui-Keun
    • The Korean Journal of Cytopathology
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    • v.7 no.1
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    • pp.1-11
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    • 1996
  • From the concepts of cellular pathology and of exfoliative cytology, as elucidated by Virchow and Papanicolaou respectively in the late 19th and early 20th century, have evolved the primary methods for the diagnosis of cancer today. From Papanicolaou's concept of exfoliative cytology developed fine needle aspiration biopsy in the early 1960's, this has become a major diagnostic procedure and has contributed to a significant reduction in open biopsies and, therefore, to medical cost-effectiveness immunobiochemical techniques provided us with a supplement to cancer diagnosis in the 1980's. The immunoperoxidase method, using monoclonal antibodies, is applied primarily as an ancillary measure to elucidate the nature of cancers The availability of specific monoclonal antibodies has greatly facilitated the identification of cell products or surface markers. For example, antibodies directed against intermediate filaments have proved to be of value in determining the histogenesis oi poorly differentiated neoplasms. Tumor markers may serve as biochemical indicators of the presence of a neoplasm. They can be detected In plasma and other body fluids. Their concentration can be applied as a diagnostic test, for monitoring the clinical course of known cancer, and as a screening measure to detect certain cancers in a population at risk. Flow cytometry is a useful tool for distinguishing several cell characteristics, such as the immunophenotype of leukemia-lymphoma cells, the DNA content of neoplastic cells, and cell proliferation rate. Molecular biologic techniques provided a giant step for the management of cancer patients encompassing diagnosis, prognostic evaluation, and therapy. Nucleic acid hybridization techniques are utilized as Southern, Northern, and dot blots and in situ hybridization. Molecular biology and its techniques may bring a blight new horizon for understanding cancer biology and in designing therapy on the basis of gene manipulation.

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