• Title/Summary/Keyword: Single-Cell Analysis

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Sensitivity and Scoring of AutoPap 300 QC System for Abnormal Cervicovaginal Cytology (비정상 자궁경부도말에서 AutoPap 300 QC System의 민감도와 Score에 영향을 주는 인자의 평가)

  • Hong, Sung-Ran
    • The Korean Journal of Cytopathology
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    • v.9 no.2
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    • pp.139-146
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    • 1998
  • The AutoPap 300 QC System is an automated device for the analysis and classification of conventional cervical cytology slides for quality control purpose. These studies evaluated the sensitivity of the AutoPap 300 QC System, and estimated morphologic features other than epithelial abnormality to identify a high quality control(QC) score with the AutoPap 300 QC System. The sensitivity of the AutoPap 300 QC System at 10% review rate for 210 cases of cervicovaginal cytology with low grade squamous intraepithelial lesion(LSIL) and higher grade lesion was assessed, and compared with a 10% random rescreening. The morphologic features, such as presence of endocervical component, dirty background, atrophy, abnormal ceil size, and celluiarity of single atypical cells were estimated in 45 cases of no review and 30 cases of QC review cases. The AutoPap 300 QC System identified 119(56.7%) out of 210 cases with LSIL and higher grade lesion at 10% review rate. It was more sensitive to squamous cell lesions$(50{\sim}62%)$ than to glandular lesions(10%). The dirty background and the scanty cellularity of single atypical cells were significantly related to low QC score. Conclusively, AutoPap 300 QC System is superior to human random rescreen for the identification of false negative smears. The upgrading of this device is required to enhance the defection of glandular lesion and certain Inadequate conditions of the slides.

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Isolation and Characterization of Nonylphenol-degrading Bacteria

  • Yu, Dae-Ung;Kim, Dong-Myung;Chung, Yong-Hyun;Lee, Yang-Bong;Kim, Young-Mog
    • Fisheries and Aquatic Sciences
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    • v.15 no.2
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    • pp.91-97
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    • 2012
  • To isolate a nonylphenol (NP)-degrading bacterium, we isolated a single colony from the NP-degrading microbial consortium SW-3, which was previously isolated from an aqueous environment. Ten colonies that exhibited different cell morphologies were isolated and the strains were named SW-3-A, -B, -C, -D, -E, -F1, -F2, -G, -H, and -I. The ability of isolates to degrade NP was evaluated by kinetic analysis by the constant of NP degradation rate ($k_1$) and the half-life time of NP degradation ($t_{1/2}$). SW-3-F1, -F2, -G, and -I strains were superior at degrading NP. The $k_1$ and $t_{1/2}$ values of the four strains were sixfold higher and one-sixth lower, respectively, than those of the consortium strain. Additionally, SW-3-F1, -G, and -I strains were tested for their ability to degrade NP during coculture. NP degradation by coculture with a combination of all three strains was inferior to that of culture conducted with single isolates, suggesting that the three strains are antagonistic toward each other during NP degradation.

The Effects of Intracellular Monocarboxylates on the ATP-sensitive Potassium Channels in Rabbit Ventricular Myocytes

  • Kim, Na-Ri;Han, Jin;Kim, Eui-Yong;Ho, Won-Kyung;Earm, Yung E.
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.5
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    • pp.581-589
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    • 1998
  • A regulating mechanism of the ATP-sensitive potassium channels $(K_{ATP}\;channels)$ is yet to fully explained. This study was carried out to investigate the effects of intracellular application of monocarboxylates (acetate, formate, lactate, and pyruvate) on $K_{ATP}$ channels in isolated rabbit ventricular myocytes. Single channel currents of $K_{ATP}$ channels were recorded using the excised inside-out or permeabilized attached (open-cell) patch-clamp technique at room temperature. Intracellular application of acetate, formate and pyruvate led to an inhibition of channel activity, whereas intracellular application of lactate increased channel activity. These effects were reversible upon washout. Analysis of single channel kinetics showed that monocarboxylates did not affect open-time constant and close-time constant. These results suggest that monocarboxylates participate in modulating $K_{ATP}$ channels activity in cardiac cells and that modulation of $K_{ATP}$ channels activity may resolve the discrepancy between the low $K_i$ in excised membrane patches and high levels of intracellular ATP concentration during myocardial ischemia or hypoxia.

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Structural Analysis of 2-Benzyl-3-[3-(4-bromo-phenyl)-1-phenyl-1H-pyrazol-4yl]-4,6-dioxo-5-phenyl-octahydro-pyrrolo[3,4-C]pyrrole-1-carboxylic Acid Ethyl Ester through X-ray Crystallography

  • Ganapathy, Jagadeesan;Pramesh, M.;Perumal, P.T.;Sanmargam, Aravindhan
    • Journal of Integrative Natural Science
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    • v.8 no.3
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    • pp.192-203
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    • 2015
  • In view of the growing medicinal importance of pyrazole and its derivatives, the single crystal X-ray diffraction study was carried out for the potential active 2-Benzyl-3-[3-(4-bromo-phenyl)-1-phenyl-1H-pyrazol-4yl]-4,6-dioxo-5-phenyl-octahydro-pyrrolo[3,4-C]pyrrole-1-carboxylic acid ethyl ester ($C_{37}H_{31}BrN_4O_4$, H2O). In the title compound are two molecules exist in the asymmetric unit. It crystallizes in the monoclinic space group $P{\hat{i}}$ with unit cell dimension $a=13.361(18){\AA}$, $b=13.424(17){\AA}$ and $c=21.649(2){\AA}$ [${\alpha}=80.745(9)^{\circ}$, ${\beta}=79.770(10)^{\circ}$ and ${\gamma}=60.788(6)^{\circ}$]. The pyrazole ring adopts planar conformation. The sum of the bond angles at nitrogen atom of the pyrazole ring indicates the $Sp^2$ hybridized state. The crystal structure is stabilized by intramolecular C-H...O hydrogen bond interaction.

Device-to-Device Relay Cooperative Transmission Based on Network Coding

  • Wang, Jing;Ouyang, Mingsheng;Liang, Wei;Hou, Jun;Liu, Xiangyang
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.11 no.7
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    • pp.3431-3445
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    • 2017
  • Due to the advantages of low transmit power consumption, high spectral efficiency and extended system coverage, Device-to-Device (D2D) communication has drawn explosive attention in wireless communication field. Considering that intra-cell interference caused between cellular signals and D2D signals, in this paper, a network coding-based D2D relay cooperative transmission algorithm is proposed. Under D2D single-hop relay transmission mode, cellular interfering signals can be regarded as useful signals to code with D2D signals at D2D relay node. Using cellular interfering signals and network coded signals, D2D receiver restores the D2D signals to achieve the effect of interference suppression. Theoretical analysis shows that, compared with Amplify-and-forward (AF) mode and Decode-and-forward (DF) mode, the proposed algorithm can dramatically increase the link achievable rate. Furthermore, simulation experiment verifies that by employing the proposed algorithm, the interference signals in D2D communication can be eliminated effectively, and meanwhile the symbol error rate (SER) performance can be improved.

Analysis of temperature-dependent abnormal bursting patterns of neurons in Aplysia

  • Hyun, Nam Gyu;Hyun, Kwangho;Oh, Saecheol;Lee, Kyungmin
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.4
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    • pp.349-362
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    • 2020
  • Temperature affects the firing pattern and electrical activity of neurons in animals, eliciting diverse responses depending on neuronal cell type. However, the mechanisms underlying such diverse responses are not well understood. In the present study, we performed in vitro recording of abdominal ganglia cells of Aplysia juliana, and analyzed their burst firing patterns. We identified atypical bursting patterns dependent on temperature that were totally different from classical bursting patterns observed in R15 neurons of A. juliana. We classified these abnormal bursting patterns into type 1 and type 2; type 1 abnormal single bursts are composed of two kinds of spikes with a long interspike interval (ISI) followed by short ISI regular firing, while type 2 abnormal single bursts are composed of complex multiplets. To investigate the mechanism underlying the temperature dependence of abnormal bursting, we employed simulations using a modified Plant model and determined that the temperature dependence of type 2 abnormal bursting is related to temperature-dependent scaling factors and activation or inactivation of potassium or sodium channels.

Large-Circular Single-stranded Sense and Antisense DNA for Identification of Cancer-Related Genes (장환형 단일가닥 DNA를 이용한 암세포 성장 억제 유전자 발굴)

  • Bae, Yun-Ui;Moon, Ik-Jae;Seu, Young-Bae;Doh, Kyung-Oh
    • Microbiology and Biotechnology Letters
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    • v.38 no.1
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    • pp.70-76
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    • 2010
  • The single-stranded large circular (LC)-sense DNA were utilized as probes for DNA chip experiments. The microarray experiment using LC-sense DNA probes found differentially expressed genes in A549 cells as compared to WI38VA13 cells, and microarray data were well-correlated with data acquired from quantitative real-time RT-PCR. A 5K LC-sense DNA microarray was prepared, and the repeated experiments and dye swap test showed consistent expression patterns. Subsequent functional analysis using LC-antisense library of overexpressed genes identified several genes involved in A549 cell growth. These experiments demonstrated proper feature of LC-sense molecules as probe DNA for microarray and the potential utility of the combination of LC-sense microarray and antisense libraries for an effective functional validation of genes.

Numerical Analysis for Two-Dimensional Compressible and Two-Phase Flow Fields of Air-Water in Eulerian Grid Framework (2차원 압축공기-물의 압축성 이상 유동 수치 해석)

  • Park, Chan-Wook;Lee, Sung-Su
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.32 no.6
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    • pp.429-445
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    • 2008
  • Two-phase compressible flow fields of air-water are investigated numerically in the fixed Eulerian grid framework. The phase interface is captured via volume fractions of each phase. A way to model two phase compressible flows as a single phase one is found based on an equivalent equation of states of Tait's type for a multiphase cell. The equivalent single phase field is discretized using the Roe‘s approximate Riemann solver. Two approaches are tried to suppress the pressure oscillation phenomena at the phase interface, a passive advection of volume fraction and a direct pressure relaxation with the compressible form of volume fraction equation. The direct pressure equalizing method suppresses pressure oscillation successfully and generates sharp discontinuities, transmitting and reflecting acoustic waves naturally at the phase interface. In discretizing the compressible form of volume fraction equation, phase interfaces are geometrically reconstructed to minimize the numerical diffusion of volume fraction and relevant variables. The motion of a projectile in a water-filled tube which is fired by the release of highly pressurized air is simulated presuming the flow field as a two dimensional one, and several design factors affecting the projectile movement are investigated.

Recent Advanced Toxicological Methods for Environmental Hazardous Chemicals (환경 오염물질의 진보된 독성 평가 기법)

  • 류재천;최윤정;김연정;김형태;방형애;송윤선
    • Environmental Analysis Health and Toxicology
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    • v.14 no.1_2
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    • pp.1-12
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    • 1999
  • Recently, several new methods for the detection of genetic damages in vitro and in vivo based on molecular biological techniques were introduced according to the rapid progress in toxicology combined with cellular and molecular biology. Among these methods, mouse lymphoma thymidine kanase (tk) gene forward mutation assay, single cell gel electrophoresis (comet assay) and transgenic animal and cell line model as a target gene of lac I (Big Blue) and lac Z (Muta Mouse) gene mutation are newly introduced based on molecular toxicological approaches. The mouse lymphoma tk$\^$+/-/ gene assay (MOLY) using L5178Y tk$\^$+/-/ mouse lymphoma cell line is one of the mammalian forward mutation assays, and has many advantages and more sensitive than hprt assay. The target gene of MOLY is a heterozygous tk$\^$+/-/ gene located in 11 chromosome, so it is able to detect the wide range of genetic changes like point mutation, deletion, rearrangement, and mitotic recombination within tk gene or deletion of entire chromosome 11. The comet assay is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakages in mammalian cells, Also, transgenic animal and cell line models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease process, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Also in vivo acridine orange supravital staining micronucleus assay by using mouse peripheral reticulocytes was introduced as an alternative of bone marrow micronucleus assay. In this respect, there was an International workshop on genotoxicity procedure (IWGTP) supported by OECD and EMS (Environmental Mutagen Society) at Washington D. C. in March 25-26, 1999. The objective of IWGTP is to harmonize the testing procedures internationally, and to extend to finalization of OECD guideline, and to the agreement of new guidelines under the International Conference of Harmonization (ICH) for these methods mentioned above. Therefore, we introduce and review the principle, detailed procedure, and application of MOLY, comet assay, transgenic mutagenesis assay and supravital staining micronucleus assay.

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Silencing of CaCDPK4 ( Capsicum annuum Calcium Dependent Protein Kinase) and ItsOrtholog, NbCDPK5 Induces Cell Death in Nicotiana benthamiana

  • Eunsook Chung;Kim, Young-Cheol;Oh, Sang-Keun;Younghee Jung;Kim, Soo-Yong;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.77.1-77
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    • 2003
  • We have isolated a full-length cDNA clone, CaCDPK4 encoding a typical calcium-dependent protein kinase (CDPK) from hot pepper cDNA library. Genomic southern blot analysis showed that it belongs to a multigene family, but represents a single copy gone in hot pepper genome. RNA expression pattern of this gene revealed that it is induced by infiltration of Xanthomonas axonopodis pv. glycines Bra into hot pepper leaves but not by water deficit stress. However, high salt treatment of NaCl (0.4 M) solution to hot pepper plants strongly induced CaCDPK4 gene. In addition, this gene is weakly responsive to the exogenous application of salicylic acid or ethephon. Biochemical study of the GST-CaCDPK4 recominant protein showed that it autophosphorylates in vitro and the presence of EGTA, a calcium chelater, eliminates the kinase activity of the recombinant protein. As a way to identify the in vivo function of CaCDPK4 in plants, VIGS (Virus-Induced Gene Silencing) was employed. Agrobacterium-mediated TRV silencing construct containing the kinase and calmodulin domain of CaCDPK4 resulted in cell death of Nicotiana benthamiana plants. A highly homologous H benthamiana CDPK gene, NbCDPK5, to CaCDPK4 was cloned from N. benthamiana cDNA library. VIGS of NbCDPK5 also resulted in cell death. The molecular characterization of this cell death phenotype is being under investigation.

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