• Title/Summary/Keyword: positive matrix

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Generation of a monoclonal anti-human $\beta$2-adrenergic receptor antibody using GST-$\beta$-adrenergic receptor C-terminal fusion proteins expressed in E.Coli.

  • Kang, Suk-Jo;Shin, Chan-Young;Park, Kyu-Hwan;Ko, Kwang-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.95-95
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    • 1997
  • Among the various receptor molecules discovered so far the ${\beta}$2-adrenergic receptors have been regarded as excellent model systems for the so called 7 transmembrane helix receptor and have been the focus of extensive studies. For the analysis of receptor structure and function a monoclonal antibody plays a crucial role, thus providing useful tools for the study of receptor. However, because of the minute quantity of receptor molecules which could be obtained from natural sources, the generation of specific monoclonal antibody against receptor molecules from the purified receptors has been regarded as virtually impractical in consideration of cost and experimental times. The purpose of the present study was to generate and characterize a monoclonal antibody against human ${\beta}$2-adrenergic receptor. For the production of antibody, C-terminal regions of the human ${\beta}$2-adrenergic receptor was produced as a fusion protein with Glutathion S-transferase (GST) in E. Coli. The expression of the fusion protein was identified by SDS-PAGE and Western blot using monoclonal anti-GST antibody. The fusion protein was purified to an apparent homogeniety by affinity chromatography with Glutathion Sepharose CL-4B and used as an antigen for the immunization of BALB/C mice. The Production of monoclonal antibody was achieved by fusion of the immunized spleen cells and SP/2-0 myeloma cells. Positive hybridomas were screened by ELISA and were cloned by two consecutive rounds of limiting dilution. The monoclonal antibody produced in this study (mAb${\beta}$C02) was IgM type and purified by immunoaffinity chromatography using anti-mouse IgM agarose as an affinity matrix. MAb${\beta}$C02 showed strong and specific immunoreactivity against both the fusion protein and human ${\beta}$2-adrenergic receptor in ELISA and Western blot. The molecular weight of immunoreactive band was 64 kDa and exactly coincided with the previously reported molecular weight of ${\beta}$2-adrenergic recepters. The results of the present study suggest that mAb${\beta}$C02 may be used for the study of receptor function and regulation in normal or nonphysiological status.

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Geochemical Analysis and Fates of Pathogenic Indicating Bacteria on Seawater Intrusion in a Sand Box Model (인공 대수층내에서 발생하는 해수침투의 지화학적 분석 및 병원성 지표 미생물의 사멸 특성)

  • Lee, So-Jung;Park, Hun-Ju;Sung, Eun-Hae;So, Myung-Ho;Kim, Chang-Gyun
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.4
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    • pp.385-392
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    • 2008
  • In this study, seawater intrusion was assessed employing a kind of biological parameters such as Escherichia coli and Enterococcus faecalis while lab-prepared reclaimed water was recharged to prevent seawater intrusion. Chemical factors indicating seawater intrusion such as Cl$^-$, Ca$^{2+}$, Mg$^{2+}$ and specific conductivity were also simultaneously investigated where an ion exchange between a matrix in artificial aquifer and cations in solution was estimated. Both Escherichia coli and Enterococcus faecalis were shown to be very sensitive against degree of salinity during saline water intrusion. Enterococcus faecalis more strongly resisted against salinity than that of Escherichia coli. The ratio of Enterococcus faecalis divided by E. coli in the process of seawater intrusion increased up to more than 50$\sim$100 times in 18 hours whereas E. coli was died off more than 90% during pumping and recharge rate kept at 10 mL/min. However, when the rates of both recharge and pumping was kept at 5 mL/min, Enterococcus faecalis / Escherichia coli was sustained in the range of 2.5$\sim$5.0, while Escherichia coli showed dimished death rate. Chemical factors such as Cl$^-$, Ca$^{2+}$, Mg$^{2+}$ and specific conductivity showed more than 0.9 of high correlation each other well explaining the degree of seawater intrusion. The degree of ion exchange between artificial aquifer and saline water can be efficiently interpreted by both minus $\Delta$Na, $\Delta$Mg variation and positive $\Delta$Ca variation.

Factor Analyses for Water Quality Indicators of Streams, Ground Water, and Reservoir in Agricultural Small Catchments of the Han River Basin

  • Park, C-S;Joo, J-H;Jung, Y-S;Yang, J-E
    • Korean Journal of Environmental Agriculture
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    • v.19 no.5
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    • pp.382-393
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    • 2000
  • The principal indicators contributing to water qualities was screened by factor analyses, based on the monitored chemical parameters of water quality for various water resources from 1995 to 1999 in the small agricultural catchments of the Han River Basin. Water samples of streams, groundwaters, and reservoirs were taken four times a year from upper (Daegwanryong), middle (Dunnae and Chunchon) and lower (Guri) reaches of Han River Basin. In these areas, the respective type of farming practiced was alpine agriculture and livestocks raising, typical upland and paddy cultivation, and intensive cropping in the plastic film house. Water quality was monitored for twenty-one water quality parameters, including pH, EC, SS, T-N, T-P, COD, cations, anions, and heavy metals. pH, EC and COD of the stream waters were suitable for the Korea irrigation water quality guidelines. However, T-N and T-P concentrations of water samples in four catchments far exceeded the irrigation water guideline. Concentrations of canons and heavy metals in Wangsuk stream in Guri area were higher than those in streams in other areas. Factor analysis revealed that significant correlation was observed for 81 pairs out of 231 water quality indicators of stream water among the $21\;{\times}\;21$ cross correlation matrix of stream water quality indicators. The first factor accounted for 27.01% of the total variation in stream water quality indicators, and high positive factor loadings were shown on EC, K, Na, $NH_4\;^+-N$, $PO_4\;^{3-}$, $SO_4\;^{2-}$, and COD. Fifty-three water quality indicator pairs were significant out of 190 ground water quality parameters. The first factor accounted for 28.54% of the total variation in ground water quality indicators, and high loadings were revealed on EC, Ca, Mg, K, Na, $NH_4\;^+-N$, and $SO_4$. Twenty-nine pairs of reservoir water quality indicators were significant out of 66 pairs. The first factor accounted for 37.06% of the total variation in reservoir water quality indicators, and high loadings were shown on EC, Mg, K, Na, SS, T-P, Cl, and COD. These results demonstrate that EC was the first factor contributing to water quality.

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Multiresidue Analysis of 124 Pesticides in Soils with QuEChERS extraction and LC-MS/MS (QuEChERS 및 LC-MS/MS를 이용한 토양 중 124종 잔류농약다성분 분석법)

  • Gwon, Ji-Hyeong;Kim, Taek-Kyum;Seo, Eun-Kyung;Hong, Su-Myeong;Kwon, Hye-Yong;Kyung, Ki-Sung;Kim, Jang-Eok;Cho, Nam-Jun
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.296-313
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    • 2014
  • A QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) multiresidue method was developed for the simultaneous analysis of 124 pesticides in soil by LC-MS. The procedure involved liquid extraction of soil immersed with 0.2N $NH_4Cl$ by acetonitrile with 1% acetic acid, followed by anhydrous $MgSO_4$ and sodium acetate, and dispersive SPE cleanup with $MgSO_4$, primary secondary amine (PSA) and $C_{18}$. The extracts were analyzed with LC-MS/MS in ESI positive mode. Standard calibration curves were made by matrix matched standards and their correlation coefficients were higher than 0.99. Recovery studies for the validation were carried out using two type soils, loam and sandy loam, at four concentration levels (0.005, 0.01, 0.02, and 0.1 mg/kg). The recoveries of pesticides were in the range of 70-120% with < 20% RSD except 4 pesticides, Benfuracarb, Ethiofencarb, Pymetrozine, and Pyrethrin. This result indicated that the method using QuEChERS and LC-MS/MS could be applied for the simultaneous determination of pesticide residues in soils.

Reduction Factors of Pesticides with Different Physicochemical Properties under Washing and Cooking Conditions (물리화학적 특성이 다른 농약의 세척 및 조리방법에 의한 감소계수 산출)

  • You, Young-Hwa;Lee, Young-Sun;Kwon, Hoon-Jeong
    • Korean Journal of Food Science and Technology
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    • v.43 no.5
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    • pp.537-543
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    • 2011
  • The reduction factor of pesticides is getting more crucial these days. However, most studies have focused on the relationship between pesticides and commodities. This study was conducted to examine the pesticide reduction factor based on their physicochemical properties. Five pesticides were selected among 288 insecticides by considering the presence of an ionizable group, the log P, and boiling points. The correlation coefficients between log P and removal by tap water, 5% acetic acid, 20% ethanol, and 0.15% detergent were -0.835 (p<0.001), 0.336 (p=0.221), 0.659 (p<0.01), and -0.939 (p<0.001), respectively. Removal by blanching was affected by log P as it showed a positive correlation with a log P of 0.620 (p<0.05). Removal by frying showed a strong negative correlation with a log P of -0.913 (p<0.001). The results suggest that removing pesticides was affected largely by log P and by vapor pressure during cooking, whereas during washing, the matrix of the food also contributed to the reduction.

Effects of Heparin-Binding Epidermal Growth Factor on the Expression of MMP-9 and ATPase ${\gamma}$-subunit mRNA in the Mouse Embryo (생쥐 배아에서 Heparin-Binding Epidermal Growth Factor가 MMP-9과 ATPase ${\gamma}$-subunit 유전자의 발현에 미치는 영향)

  • Hong, Seok-Ho;Nah, Hee-Young;Lee, Ji-Yun;Kim, Jung-Hee;Kim, Chung-Hoon;Chae, Hee-Dong;Kang, Byung-Moon;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.2
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    • pp.87-93
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    • 2001
  • Objectives: The purpose of this study was to evaluate effects of heparin-binding epidermal growth factor (HB-EGF) on the rate of blastocyst formation and hatching in the mouse embryos and the expression of matrix metalloproteinase-9 (MMP-9) and ATPase ${\gamma}$-subunit mRNA. Methods: Late 2-cell mouse embryos was cultured for 72 hours in RTF medium containing with 1, 10, and 100 ng/ml HB-EGF. The mRNA expression level of MMP-9 and ATPase ${\gamma}$-subunit was detected by reverse transcription-polymerase chain reaction (RT-PCR). Results: The rate of hatching was significantly higher (p<0.05) in group containing with 1 ng/ml HB-EGF than other groups. Also, the rate of hatched blastocyst was significantly higher (p<0.05) in 10 ng/ml. The mRNA expression level of MMP-9 mRNA was not shown any difference among groups, but ATPase ${\gamma}$-subunit was higher than other groups. Conclusions: Taken together these results suggest that HB-EGF has the positive effect to promote the blastocyst formation and hatching process and influences the blastocoel expansion by promoting the ATPase mRNA expression in the mouse embryos.

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Art and Collectivity (미술과 집단성)

  • Kwok, Kian-Chow
    • The Journal of Art Theory & Practice
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    • no.4
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    • pp.181-202
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    • 2006
  • "When it comes to art, nationalism is a goodticket to ride with", says the title of a report in the Indian Express (Mumbai, 29 Oct 2000). The newspaper report goes on to say that since Indian art was kept "ethnic" by colonialism, national liberation meant opening up to the world on India's own terms. Advocacy, at the tail end of the 20th century, would contrast dramatically with the call by Rabindranath Tagore, the founder of the academy at Santiniketan in 1901, to guard against the fetish of nationalism. "The colourless vagueness of cosmopolitanism," Tagore pronounced, "nor thefierce self-idolatry of nation-worship, is the goal of human history" (Nationalism, 1917). This contrast is significant on two counts. First is the positive aspect of "nation" as a frame in art production or circulation, at the current point of globalization when massive expansion of cultural consumers may be realized through prevailing communication networks and technology. The organization of the information market, most vividly demonstrated through the recent FIFA World Cup when one out of every five living human beings on earth watched the finals, is predicated on nations as categories. An extension of the Indian Express argument would be that tagging of artworks along the category of nation would help ensure greatest reception, and would in turn open up the reified category of "art," so as to consider new impetus from aesthetic traditions from all parts of the world many of which hereto fore regarded as "ethnic," so as to liberate art from any hegemony of "international standards." Secondly, the critique of nationalism points to a transnational civic sphere, be it Tagore's notion of people-not-nation, or the much mo re recent "transnational constellation" of Jurgen Habermas (2001), a vision for the European Union w here civil sphere beyond confines of nation opens up new possibilities, and may serve as a model for a liberated sphere on global scale. There are other levels of collectivity which art may address, for instance the Indonesian example of local communities headed by Ketua Rukun Tetangga, the neighbourhood headmen, in which community matters of culture and the arts are organically woven into the communal fabric. Art and collectivity at the national-transnational level yield a contrasting situation of, on the idealized end, the dual inputs of local culture and tradition through "nation" as necessary frame, and the concurrent development of a transnational, culturally and aesthetically vibrant civic sphere that will ensure a cosmopolitanism that is not a "colourless vagueness." In art historical studies, this is seen, for instance, in the recent discussion on "cosmopolitan modernisms." Conversely, we may see a dual tyranny of a nationalism that is a closure (sometimes stated as "ethno-nationalism" which is disputable), and an internationalism that is evolved through restrictive understanding of historical development within privileged expressions. In art historical terms, where there is a lack of investigation into the reality of multiple modernisms, the possibility of a democratic cosmopolitanism in art is severely curtailed. The advocacy of a liberal cosmopolitanism without a democratic foundation returns art to dominance of historical privileged category. A local community with lack of transnational inputs may sometimes place emphasis on neo-traditionalism which is also a double edged sword, as re kindling with traditions is both liberating and restrictive, which in turn interplays with the push and pull of the collective matrix.

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Identification of Atmospheric PM10 Sources and Estimating Their Contributions to the Yongin-Suwon Bordering Area by Using PMF (PMF모델을 이용한 용인.수원 경계지역에서 PM10 오염원의 확인과 상대적 기여도의 추정)

  • Lee, Hyung-Woo;Lee, Tae-Jung;Yang, Sung-Su;Kim, Dong-Sool
    • Journal of Korean Society for Atmospheric Environment
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    • v.24 no.4
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    • pp.439-454
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    • 2008
  • The purpose of this study was to extensively identify $PM_{10}$ sources and to estimate their contributions to the study area, based on the analysis of the $PM_{10}$ mass concentration and the associated inorganic elements, ions, and total carbon. The contribution of $PM_{10}$ sources was estimated by applying a receptor method because identifying air emission sources were effective way to control the ambient air quality. $PM_{10}$ particles were collected from May to November 2007 in the Yongin-Suwon bordering area. $PM_{10}$ samples were collected on quartz filters by a $PM_{10}$ high-volume air sampler. The inorganic elements (Al, Mn, V, Cr, Fe, Ni, Cu, Zn, Cd, Pb, Si, Ba, Ti and Ag) were analyzed by an ICP-AES after proper pre-treatments of each sample. The ionic components of these $PM_{10}$ samples ($Cl^_$, $NO_3^-$, $SO_4^{2-}$, $Na^+$, $NH_4^+$, $K^+$, $Ca^{2+}$, and $Mg^{2+}$) were analyzed by an IC. The carbon components (OC1, OC2, OC3, OC4, OP, EC1, EC2 and EC3) were also analyzed by DRI/OGC analyzer. Source apportionment of $PM_{10}$ was performed using a positive matrix factorization (PMF) model. After performing PMF modeling, a total of 8 sources were identified and their contribution were estimated. Contributions from each emission source were as follows: 13.8% from oil combustion and industrial related source, 25.4% from soil source, 22.1% from secondary sulfate, 12.3% from secondary nitrate, 17.7% from auto emission including diesel (12.1%) and gasoline (5.6%), 3.1% from waste incineration and 5.6% from Na-rich source. This study provides information on the major sources affecting air quality in the receptor site, and therefore it will help us maintain and manage the ambient air quality in the Yongin-Suwon bordering area by establishing reliable control strategies for the related sources.

Identification of PM10 Chemical Characteristics and Sources and Estimation of their Contributions in a Seoul Metropolitan Subway Station (서울시 지하역사에서 PM10의 화학적 특성과 오염원의 확인 및 기여도 추정)

  • Park, Seul-Ba-Sen-Na;Lee, Tae-Jung;Ko, Hyun-Ki;Bae, Sung-Joon;Kim, Shin-Do;Park, Duckshin;Sohn, Jong-Ryeul;Kim, Dong-Sool
    • Journal of Korean Society for Atmospheric Environment
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    • v.29 no.1
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    • pp.74-85
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    • 2013
  • Since the underground transportation system is a closed environment, indoor air quality problems may seriously affect many passengers' health. The purpose of this study was to understand $PM_{10}$ characteristics in the underground air environment and further to quantitatively estimate $PM_{10}$ source contributions in a Seoul Metropolitan subway station. The $PM_{10}$ was intensively collected on various filters with $PM_{10}$ aerosol samplers to obtain sufficient samples for its chemical analysis. Sampling was carried out in the M station on the Line-4 from April 21 to 28, July 13 to 21, and October 11 to 19 in the year of 2010 and January 11 to 17 in the year of 2011. The aerosol filter samples were then analyzed for metals, water soluble ions, and carbon components. The 29 chemical species (OC1, OC2, OC3, OC4, CC, PC, EC, Ag, Al, Ba, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Si, Ti, V, Zn, $Cl^-$, $NO_3{^-}$, $SO_4{^{2-}}$, $Na^+$, $NH_4{^+}$, $K^+$, $Mg^{2+}$, $Ca^{2+}$) were analyzed by using ICP-AES, IC, and TOR after proper pretreatments of each sample filter. Based on the chemical information, positive matrix factorization (PMF) model was applied to identify the $PM_{10}$ sources and then six sources such as biomass burning, outdoor, vehicle, soil and road dust, secondary aerosol, ferrous, and brakewear related source were classified. The contributions rate of their sources in tunnel are 4.0%, 5.8%, 1.6%, 17.9%, 13.8% and 56.9% in order.

Effects of Tiam 1 on Invasive Capacity of Gastric Cancer Cells in vitro and Underlying Mechanisms

  • Zhu, Jin-Ming;Yu, Pei-Wu
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.201-208
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    • 2013
  • Objective: To investigate changes in the invasive capacity of gastric cancer cells in vitro after expression inhibition of T lymphoma invasion and metastasis inducing factor 1 (Tiam 1) and underlying mechanisms. Methods: Using adhesion selection, two subpopulations with high ($M_H$) or low ($M_L$) invasive capacity were separated from the human gastric cancer cell line MKN-45 ($M_0$). Tiam 1 antisense oligodeoxynucleotide (ASODN) was transfected into $M_H$ cells with liposomes, and expression of Tiam 1 mRNA and protein was determined by RT-PCR and quantitative cellular-ELISA. Changes in the cytoskeleton, invasive capacity in vitro and expression of ras-related $C_3$ botulinum toxin substrate 1 (Rac 1), integrin ${\beta}1$ and matrix metalloproteinase 2 (MMP 2) between Tiam 1 ASODN transfected $M_H$ cells and non-transfected cells were observed by HE staining, cytoskeletal protein staining, scanning electron microscopy, Boyden chamber tests and cyto-immunohistochemistry. Results: A positive correlation existed between the expression level of Tiam l mRNA or protein and the invasion capacity of gastric cancer cells. After ASODN treatment ($0.43{\mu}M$ for 48 h), Tiam 1 mRNA transcription and protein expression in $M_H$ cells were decreased by 80% and 24% respectively (P < 0.05), compared with untreated controls, while invasive capacity in vitro was suppressed by 60% (P < 0.05). Morphologic and ultrastructural observation also showed that ASODN-treated $M_H$ cells exhibited smooth surfaces with obviously reduced filopodia and microspikes, which resembled $M_0$ and $M_L$ cells. Additionally, cytoskeletal distribution dramatically altered from disorder to regularity with reduced long filament-like structure, projections, pseudopodia on cell surface, and with decreased acitn-bodies in cytoplasm. After Tiam 1 ASODN treatment, the expression of Rac 1 and Integrin ${\beta}1$ in $M_H$ cells was not affected (P > 0.05), but that of MMP 2 in $M_H$ cells was significantly inhibited compared with untreated cells (P < 0.05). Conclusion: Over-expression of Tiam-1 contributes to the invasive phenotype of gastric cancer cells. Inhibition of Tiam 1 expression could impair the invasive capacity of gastric cancer cells through modulating reconstruction of the cytoskeleton and regulating expression of MMP 2.