• Title/Summary/Keyword: Single-Cell Analysis

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$V_H$ Gene Expression and its Regulation on Several Different B Cell Population by using in situ Hybridization technique

  • Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.111-122
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    • 1993
  • The mechanism by which $V_H$ region gene segments is selected in B lymphocyte is not known. Moreover, evidence for both random and nonrandom expression of $V_H$ genes in matured B cells has been presented previously. In this report, the technique of in situ hybridization allowed us to analyze expressed $V_H$ gene families in normal B lymphocyte at the single cell level. The analysis of normal B cells in this study eliminated any posssible bias resulting from transformation protocols used previously and minimized limitation associated with sampling size. Therefore, an accurate measure of the functional and expressed $V_H$ gene repertoire in B lymphocyte could be made. One of the most important controls for the optimization of in situ hybridization is to establish probe concentration and washing stringency due to the degree of nucleotide sequence similarlity between different families which in some cases can be as high as 70%. When the radioactive $C{\mu}$ and $V_{H}J558$ RNA probes are tested on LPS-stimulated adult spleen cells, $2{\sim}4{\times}106cpm$/slide shows low background and reasonable frequency of specific positive cells. For the washing condition. 40~50% formamide at $54^{\circ}C$ is found to be optimum for the $C{\mu}$. $V_{H}S107$ and $V_{H}J558$ probes. The analyzed results clearly demonstrate that the level of each different $V_H$ gene family expression is dependent upon the complexity or size of that family. These findings are also extended to the level of $V_H$ gene family expression in separated bone marrow B cells depend upon the various stage of differentiation and conclude no preferential utilization of specific $V_H$ gene family. Thus, the utilization of VH gene segments in B lymphocyte of adult BALB/c mice is random and is not regulated or changed during the differentiation of B cells.

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A Novel type of High-Frequency Transformer Linked Soft-Switching PWM DC-DC Power Converter for Large Current Applications

  • Morimoto Keiki;Ahmed Nabil A.;Lee Hyun-Woo;Nakaoka Mutsuo
    • Journal of Electrical Engineering and Technology
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    • v.1 no.2
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    • pp.216-225
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    • 2006
  • This paper presents a new circuit topology of DC busline switch and snubbing capacitor-assisted full-bridge soft-switching PWM inverter type DC-DC power converter with a high frequency link for low voltage large current applications as DC feeding systems, telecommunication power plants, automotive DC bus converters, plasma generator, electro plating plants, fuel cell interfaced power conditioner and arc welding power supplies. The proposed power converter circuit is based upon a voltage source-fed H type full-bridge high frequency PWM inverter with a high frequency transformer link. The conventional type high frequency inverter circuit is modified by adding a single power semiconductor switching device in series with DC rail and snubbing lossless capacitor in parallel with the inverter bridge legs. All the active power switches in the full-bridge inverter arms and DC busline can achieve ZVS/ZVT turn-off and ZCS turn-on commutation operation. Therefore, the total switching losses at turn-off and turn-on switching transitions of these power semiconductor devices can be reduced even in the high switching frequency bands ranging from 20 kHz to 100 kHz. The switching frequency of this DC-DC power converter using IGBT power modules is selected to be 60 kHz. It is proved experimentally by the power loss analysis that the more the switching frequency increases, the more the proposed DC-DC converter can achieve high performance, lighter in weight, lower power losses and miniaturization in size as compared to the conventional hard switching one. The principle of operation, operation modes, practical and inherent effectiveness of this novel DC-DC power converter topology is proved for a low voltage and large current DC-DC power supplies of arc welder applications in industry.

Functional Effects of Increased Copy Number of the Gene Encoding Proclavaminate Amidino Hydrolase on Clavulanic Acid Production in Streptomyces clavuligerus ATCC 27064

  • Song, Ju-Yeon;Kim, Eun-Sook;Kim, Dae-Wi;Jesen, Susan E.;Lee, Kye-Joon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.417-426
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    • 2008
  • The effect of increasing levels of proclavaminate amidino hydrolase (Pah) on the rate of clavulanic acid production in Streptomyces clavuligerus ATCC 27064 was evaluated by increasing dosoge of a gene (pah2) encoding Pah. A strain (SMF5703) harboring a multicopy plasmid containing the pah2 gene showed significantly retarded cell growth and reduced clavulanic acid production, possibly attributable to the deleterious effects of the multicopy plasmid. In contrast, a strain (SMF5704) carrying a single additional copy of pah2 introduced into chromosome via an integrative plasmid showed enhanced production of clavulanic acid and increased levels of pah2 transcripts. Analysis of transcripts of other genes involved in the clavulanic acid biosynthetic pathway revealed a pattern similar to that seen in the parent. From these results, it appears that clavulanic acid production can be enhanced by duplication of pah2 through integration of a second copy of the gene into chromosome. However, increasing the copy number of only one gene, such as pah2, does not affect the expression of other pathway genes, and so only modest improvements in clavulanic acid production can be expected. Flux controlled by Pah did increase when the copy number of pah2 was doubled, suggesting that under these growth conditions, Pah levels may be a limiting factor regulating the rate of clavulanic acid biosynthesis in S. clavuligerus.

Sintering Behavior and Electrical Properties of Strontium Titanate-Based Ceramic Interconnect Materials for Solid Oxide Fuel Cells (고체산화물 연료전지용 Strontium Titanate 세라믹 접속자 소재의 소결 거동 및 전기적 특성)

  • Park, Beom-Kyeong;Lee, Jong-Won;Lee, Seung-Bok;Lim, Tak-Hyoung;Park, Seok-Joo;Song, Rak-Hyun;Shin, Dong-Ryul
    • 한국신재생에너지학회:학술대회논문집
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    • 2010.11a
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    • pp.80.1-80.1
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    • 2010
  • A strontium titanate ($SrTiO_3$)-based material with a perovskite structure is considered to be one of the promising alternatives to $LaCrO_3$-based materials since $SrTiO_3$ perovskite shows a high chemical stability under both oxidizing and reducing atmospheres at high temperatures. $SrTiO_3$ materials exhibit an n-type semiconducting behavior when it is donor-doped and/or exposed to a reducing atmosphere. In this work, $Sr_{1-x}La_xTi_{1-y}M_yO_3$ materials doped with $La^{3+}$ in A-sites and aliovalent transition metal ions ($M^{n+}$) in B-sites were synthesized by the modified Pechini method. The X-ray diffraction analysis indicated that the materials synthesized by the Pechini process exhibited a single curbic perovskite-type structure without any impurity phases, and are tolerant, to some extent, to cation doping. The sintering behaviors of $Sr_{1-x}La_xTi_{1-y}M_yO_3$ in $H_2/N_2$ and air were characterized by dilatometry and microstructural observations. The electrical conduction mechanism and the dopant effect are discussed based on the defect structures and the electrical conductivities measured at various oxygen partial pressures and temperatures.

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Effect of Deer Antler Drink Supplementation on Blood Pressure, Blood Glucose and Lymphocyte DNA Damage in Type 2 Diabetic Patients (당뇨환자에 있어서 녹용추출물의 섭취가 혈압, 혈당 및 임파구 DNA 손상에 미치는 영향)

  • Kim Hye-Young;Jeon Eun-Jae;Park Yoo Kyoung;Kang Myung-Hee
    • Journal of Nutrition and Health
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    • v.37 no.9
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    • pp.794-800
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    • 2004
  • Deer Antler has been known for its traditional oriental medicinal properties and has been widely used to promote growth, boost immune function, treat blood loss and chronic joint pain. Recent study showed imported (New Zealand) Deer Antler was beneficial in reducing the side effects of cancer treatments. However, there was no intervention study conducted on the effect of Korean Deer Antler on reducing the oxidative stress to patients with diabetes. One of the sensitive ways to measure endogenous oxidative stress is by measuring cellular DNA damage using single cell gel electrophoresis (COMET assay). This study was conducted to investigate the possible beneficial effect of commercial Deer Antler drink (provided by Chung-yang Deer Farm) on lymphocyte DNA damage and blood glucose of diabetic patients. Ten patients (4 men, 6 women) participated in the study and consumed 2 pouches of Deer Antler drink every day for 20 days. Blood was collected on the morning before and after the intervention for lymphocyte isolation and blood glucose analysis. Both systolic and diastolic blood pressure showed a tendency to decrease but did not reach statistical significance after the trial. Blood glucose level was not affected by the supplementation. After the intervention, over 50% reduction were noted in the cellular DNA damage, expressed as tail length (TL) and tail moment (TM: tail length ${\times}$ percent tail DNA) . Although we did not obtain beneficial effect on lowering blood glucose levels in the patients, this results suggest that Deer Antler may initially act in protecting endogenous DNA damage in short-term experiment.

Analysis of two-stage Continuous Culture System by Transient Response of Single-stage Continuous Culture System (일단 연속 생물반응기의 과도상태 거동을 이용한 이단 연속 생물반응기의 해석)

  • 박성훈;공인수
    • KSBB Journal
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    • v.7 no.4
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    • pp.308-316
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    • 1992
  • Two-stage continuous culture system has been studied intensively to maximize the productivity of a cloned gene product in unstable recombinant microorganism. As an effort to optimize the two-stage process, transient behavior of the second-stage was studied theoretically as well as experimentally using Escherichia coli Kl2$\delta$Hl$\delta$trp. A mathematical model describing the transient response to a step change in dilution rate was developed based on the assumption that the adaptation rate of cell growth is proportional to the available growth potential, which is defined as the difference in dilution rates between before and after shift-up. The kinetic parameters appearing in the model equations were the dimensionless step increase in growth rate($\alpha$) and the adaptation rate constant(k). These parameters were evaluated for various dilution rates and temperatures by washout method. This relatively simple adaptation model could predict the specific growth rate of the second-stage successfully. Advantage and disadvantage of the proposed model are also discussed.

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Crystal Structures, Electrical Conductivities and Electrochemical Properties of LiCo1-XMgxO2(x=0.03) for Secondary Lithium Ion Batteries (리튬 2차 전지용 LiCo1-XMgxO2(x=0.03)의 결정구조, 전기전도도 및 전기화학적 특성)

  • Kim, Ho-Jin;Chung, Uoo-Chang;Jeong, Yeon-Uk;Lee, Joon-Hyung;Kim, Jeong-Joo
    • Journal of the Korean Ceramic Society
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    • v.42 no.9 s.280
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    • pp.602-606
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    • 2005
  • [ $LiCoO_{2}$ ] is the most common cathode electrode materials in Lithium-ion batteries. $LiCo_{0.97}Mg_{0.03}O_2$ was synthesized by the solid-state reaction method. We investigated crystal structures, electrical conductivities and electrochemical properties. The crystal structure of $LiCo_{0.97}Mg_{0.03}O_2$ was analyzed by X-ray powder diffraction and Rietveld refinement. The material showed a single phase of a layered structure with the space group R-3m. The lattice parameter(a, c) of $LiCo_{0.97}Mg_{0.03}O_2$ was larger than that of $LiCoO_2$. The electrical conductivity of sintered samples was measured by the Van der Pauw method. The electrical conductivities of $LiCoO_2$ and $LiCo_{0.97}Mg_{0.03}O_2$ were $2.11{\times}10^{-4}\;S/cm$ and $2.41{\times}10^{-1}\;S/cm$ at room temperature, respectively. On the basis of the Hall effect analysis, the increase in electrical conductivities of $LiCo_{0.97}Mg_{0.03}O_2$ is believed due to the increased carrier concentrations, while the carrier mobility was almost invariant. The electrochemical performance was investigated by coin cell test. $LiCo_{0.97}Mg_{0.03}O_2$ showed improved cycling performance as compared with $LiCoO_2$.

Novel Single-Nucleotide Polymorphisms of SOHLH2 in Korean Patients with Premature Ovarian Failure

  • Jeong, Ji-Hye;Choi, Mi-Kyung;Won, Hyung-Jae;Song, Seung-Hoon;Kim, You-Shin;Lyu, Sang-Woo;Seok, Hyun-Ha;Park, Mi-Ree;Kim, Nam-Keun;Yoon, Tae-Ki;Shim, Sung-Han;Choi, Young-Sok
    • Development and Reproduction
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    • v.15 no.4
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    • pp.309-313
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    • 2011
  • SOHLH2 is a novel germ cell-specific transcription factor that is crucial for folliculogenesis in the ovary and spermatogenesis in the testis. SOHLH2 represents a candidate gene for infertility with premature ovarian failure. We analyzed whether mutations in the SOHLH2 gene in 98 Korean women with premature ovarian failure. The sequence analysis identified six novel SNPs (c.431-41G>C, c.656A>T, c.1000+27C>T, c.1000+33G>T, c1258-106G>A, and c.2094+ 11T>C) from Korean patients with premature ovarian failure. The c.656A>T found in exon 7 results in change of an amino acid, tyrosine to phenylalanine. Functional mutations in SOHLH2 gene are rare in Korean women with premature ovarian failure.

Expressional Changes of Connexin Isoform Genes in the Rat Caput Epididymis Exposed to Flutamide or Estradiol Benzoate at the Early Postnatal Age

  • Lee, Ki-Ho
    • Development and Reproduction
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    • v.21 no.3
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    • pp.317-325
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    • 2017
  • Direct communication between neighboring cells through connexin (Cx)-based gap junction is a crucial biological manner to regulate functions of a tissue consisting of multi-cell types. The present research evaluated expressional changes of Cx isoforms in the caput epididymis of adult rat exposed to estradiol benzoate (EB) or flutamide (Flu) at the early postnatal age. A single subcutaneous administration of EB at a low-dose [$0.015{\mu}g/kg$ body weight (BW)] or a high-dose ($1.5{\mu}g/kg\;BW$) or Flu at a low-dose ($500{\mu}g/kg\;BW$) or a high-dose (5 mg/kg BW) was performed to an animal at 1 week of age. Quantitative real-time PCR analysis was employed to determine expressional changes of Cx isoforms. The transcript levels of Cxs30.3 and 37 were decreased by a low-dose EB treatment, while decreases of Cxs31, 31.1, 32, 40, and 45 transcript levels were observed with a low-dose EB treatment. The treatment of a high-dose EB resulted in expressional reduction of Cxs30.3, 31, 31.1, 37, 40, 43, and 45. The Flu treatment at a low dose caused increases of Cxs26, 37, and 40 transcript levels but decreases of Cxs31.1, 43, and 45 transcript levels. Increases of Cxs30.3, 31, 37, and 40 mRNA amounts were induced by a high-dose Flu treatment. However, exposure to a high-dose Flu produced expressional decreases of Cxs31.1, 32, and 43 in the adult caput epididymis. These observations suggest that exposure to EB or Flu at the neonatal period could lead to aberrant expression of Cx isoforms in the adult caput epididymis.

Cytologic Analysis of Malignant Tumor Cells in Cerebrospinal Fluid (뇌척수액에서 진단된 악성 종양세포의 세포학적 분석)

  • Suh, Jae-Hee;Gong, Gyung-Yub;Khang, Shin-Kwang;Kim, On-Ja
    • The Korean Journal of Cytopathology
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    • v.9 no.1
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    • pp.21-28
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    • 1998
  • Cytologic evaluation of cerebrospinal fluid(CSF) is an effective tool in diagnosing many disorders involving the central nervous system(CNS). CSF examination has been found to be of particular value in the diagnosis of metastatic carcinoma, lymphomatous or leukemic involvement of CNS and certain primary CNS tumors. As a survey of metastatic tumors to CSF and an evaluation of the preparation techniques increasing cellular yield in our laboratory, 713 CSF specimens examined between July 1995 and April 1997(1 year 10 months), were reviewed. There were 75 positive and 5 suspicious cases, the latter have had no evidence of tumors clinically. Primary tumors of 75 positive cases were classified as follows; 4(5.3%) as primary brain tumors, 40(53.3%) as secondary carcinomas, 13(17.3%) as leukemias, and 18 (24.0%) as lymphomas. The most common primary site of metastatic carcinomas was the lung in 17 cases(42.5%) followed by the stomach in 13(32.5%), breast in 8 (20.0%), and unknown primary in 2(5.0%). Four primary brain tumors were 3 cerebellar medulloblastomas and a supratentorial primitive neuroectodermal tumor (PNET). All 40 metastatic carcinomas were adenocarcinoma presented as single cells or cell clusters. Although signet ring cells were frequent in the cases of gastric primary cancers, no significant cytologic differences according to the primary site were observed. The cytologic features of leukemia and lymphoma were characterized by hypercellular smears presenting as individual atypical cells with increased N/C ratio, presence of nucleoli, and nuclear protrusions. In medulloblastomas and PNET, the principal cytologic findings were small undifferentiated cells arranged singly or in loose clusters with occasional rosettoid features. This study suggests that the CSF cytology is useful in the diagnosis of malignancy, especially metastatic extracranial tumors and the diagnostic accuracy can be improved by increasing cellular yield using cytocentrifuge.

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