• Title/Summary/Keyword: Indirect Method

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Effects of Neurotrophic Factors on the Generation of Functional Dopamine Secretory Neurons Derived from in vitro Differentiated Human Embryonic Stem Cells (신경성장촉진 인자가 인간 배아줄기세포 유래 도파민 분비 신경세포형성에 미치는 영향)

  • Lee, Keum-Sil;Kim, Eun-Young;Shin, Hyun-Ah;Cho, Hwang-Yoon;Wang, Kyu-Chang;Kim, Yong-Sik;Lee, Hoon-Taek;Chung, Kil-Saeng;Lee, Won-Don;Park, Se-Pill;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.19-27
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    • 2004
  • Objective: This study was to examine the in vitro neural cell differentiation patterns of human embryonic stem (hES) cells following treatment of various neurotrophic factors [basic fibroblast growth factor (bFGF), retinoic acid (RA), brain derived neurotrophic factor (BDNF) and transforming growth factor (TGF)-$\alpha$], particulary in dopaminergic neuron formation. Methods: The hES cells were induced to differentiate by bFGF and RA. Group I) In bFGF induction method, embryoid bodies (EBs, for 4 days) derived from hES were plated onto gelatin dish, selected for 8 days in ITSFn medium and expanded at the presence of bFGF (10 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14 and 21 days. Group II) For RA induction, EBs were exposed of RA ($10^{-6}M$) for 4 days and allowed to differentiate in N2 medium for 7, 14 and 21 days. Group III) To examine the effects of additional neurotrophic factors, bFGF or RA induced cells were exposed to either BDNF (10 ng/ml) or TGF-$\alpha$ (10 ng/ml) during the 21 days of final differentiation. Neuron differentiation and dopamine secretion were examined by indirect immunocytochemistry and HPLC, respectively. Results: The bFGF or RA treated hES cells were resulted in similar neural cell differentiation patterns at the terminal differentiation stage, specifically, 75% neurons and 11% glial cells. Additionally, treatment of hES cells with BDNF or TGF-$\alpha$ during the terminal differentiation stage led to significantly increased tyrosine hydroxylase (TH) expression of a dopaminergic neuron marker, compared to control (p<0.05). In contrast, no effect was observed on the rate of mature neuron (NF-200) or glutamic acid decarboxylase-positive neurons. Immunocytochemistry and HPLC analyses revealed the higher levels of TH expression (20.3%) and dopamine secretion (265.5 $\pm$ 62.8 pmol/mg) in bFGF and TGF-sequentially treated hES cells than those in $\alpha$ RA or BDNF treated hES cells. Conclusion: These results indicate that the generation of dopamine secretory neurons from in vitro differentiated hES cells can be improved by TGF-$\alpha$ addition in the bFGF induction protocol.

In vitro Neural Cell Differentiation of Genetically Modified Human Embryonic Stem Cells Expressing Tyrosine Hydroxylase (Tyrosine Hydroxylase 유전자가 주입된 인간 배아줄기세포의 체외 신경세포 분화)

  • Shin, Hyun-Ah;Kim, Eun-Young;Lee, Keum-Sil;Cho, Hwang-Yoon;Kim, Yong-Sik;Lee, Won-Don;Park, Se-Pill;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.67-74
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    • 2004
  • Objective: This study was to examine in vitro neural cell differentiation pattern of the genetically modified human embryonic stem cells expressing tyrosine hydroxylase (TH). Materials and Methods: Human embryonic stem (hES, MB03) cell was transfected with cDNAs cording for TH. Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA, embryoid bodies (EB, for 4 days) derived from TH#2/MB03 cells were exposed to RA ($10^{-6}M$)/AA ($5{\times}10^{-2}mM$) for 4 days, and were allowed to differentiate in N2 medium for 7, 14 or 21 days. Exp. II) When b-FGF was used, neuronal precursor cells were expanded at the presence of b-FGF (10 ng/ml) for 6 days followed by a final differentiation in N2 medium for 7, 14 or 21 days. Neuron differentiation was examined by indirect immunocytochemistry using neuron markers (NF160 & NF200). Results: After 7 days in N2 medium, approximately 80% and 20% of the RA or b-FGF induced Th#2/MB03 cells were immunoreactive to anti-NF160 and anti-NF200 antibodies, respectively. As differentiation continued, NF200 in RA treated cells significantly increased to 73.0% on 14 days compared to that in b-FGF treated cells (53.0%, p<0.05), while the proportion of cells expressing NF160 was similarly decreased between two groups. However, throughout the differentiation, expression of TH was maintained ($\sim$90%). HPLC analyses indicated the increased levels of L-DOPA in RA treated genetically modified hES cells with longer differentiation time. Conclusion: These results suggested that a genetically modified hES cells (TH#2/MB03) could be efficiently differentiated in vitro into mature neurons by RA induction method.

Changes in Mortality Inequality in Relation to the South Korean Economic Crisis: Use of Area-based Socioeconomic Position (경제위기에 따른 사망률 불평등의 변화: 지역의 사회경제적 위치 지표의 활용)

  • Yun, Sung-Cheol;Hwang, In-A;Lee, Moo-Song;Lee, Sang-Il;Jo, Min-Woo;Lee, Min-Jung;Khang, Young-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.38 no.3
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    • pp.359-365
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    • 2005
  • Objectives : An abrupt economic decline may widen the socioeconomic differences in health between the advantaged and disadvantaged in a society. The aim of this study was to examine whether the South Korean economic crisis of 1997-98 affected the socioeconomic inequality from all-causes and from cause-specific mortality between 1995 and 2001. Methods : Population denominators were obtained from the registration population data, with the number of death (numerators) calculated from raw death certificate data. The indicator used to assess the geographic socioeconomic position was the per capita regional tax revenue. Administrative districts (Si-Gun-Gu) were ranked according to this socioeconomic measure, and divided into equal population size quintiles on the basis of this ranking. The sex- and 5-year age-specific numbers of the population and deaths were used to compute the sex- and age-adjusted mortality rates (via direct standardization method), standardized mortality ratios (via indirect standardization methods) and relative indices of inequality (RII) (via Poisson regression). Results : Geographic inequalities from all-causes of mortality, as measured by RII, did not increase as a result of the economic crisis (from 1998-2001). This was true for both sexes and all age groups. However, the cause-specific analyses showed that socioeconomic inequalities in mortalities from external causes were affected by South Korean economic crisis. For males, the RIIs for mortalities from transport accidents and intentional self-harm increased between 1995 and 2001. For females, the RII for mortality from intentional self-harm increased during the same period. Conclusions : The South Korean economic crisis widened the geographic inequality in mortalities from major external causes. This increased inequality requires social discourse and counter policies with respect to the rising health inequalities in the South Korean society.

Microfluidic Immuno-Sensor Chip using Electrical Detection System (전기 검출 시스템을 이용한 Microfluidic Immuno-Sensor Chip)

  • Maeng, Joon-Ho;Lee, Byung-Chul;Cho, Chul-Ho;Ko, Yong-Jun;Ahn, Yoo-Min;Cho, Nahm-Gyoo;Lee, Seoung-Hwan;Hwang, Seung-Yong
    • KSBB Journal
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    • v.21 no.5
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    • pp.325-330
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    • 2006
  • This study presents the characterization of an integrated portable microfluidic electrical detection system for fast and low volume immunoassay using polystyrene microbead, which are used as immobilization surfaces. In our chip, a filtration method using the microbead was adopted for sample immobilization and immunogold silver staining(IGSS) was used to increase the electrical signal. The chip is composed of an inexpensive and biocompatible Polydimethylsiloxane(PDMS) layer and Pyrex glass substrate. Platinum microelectrodes for electric signal detection were fabricated on the substrate and microchannel and pillar-type microfilters were formed in the PDMS layer. With a fabricated chip, we reacted antigen and antibody according to the procedures. Then, silver enhancer was injected to increase the size of nanogold particles tagged with the second antibody. As a result, microbeads were connected to each other and formed an electrical bridge between microelectrodes. Resistance measured through the electrodes showed a difference of two orders of magnitude between specific and nonspecific immuno-reactions. The detection limit was 10 ng/ml. The developed immunoassay chip reduced the total analysis time from 3 hours to 50 min. Fast and low-volume biochemical analysis has been successfully achieved with the developed microfilter and immuno-sensor chip, which is integrated to the microfluidic system.

Life Cycle Cost Analysis at Design Stage of Cable Stayed Bridges based on the Performance Degradation Models (성능저하모델에 기초한 사장교의 설계단계 생애주기비용 분석)

  • Koo, Bon Sung;Han, Sang Hoon;Cho, Choong Yuen
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.33 no.5
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    • pp.2081-2091
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    • 2013
  • Recently, the demand on the practical application of life-cycle cost effectiveness for design and rehabilitation of civil infrastructure is rapidly growing unprecedently in civil engineering practice. Accordingly, in the 21st century, it is almost obvious that life-cycle cost together with value engineering will become a new paradigm for all engineering decision problems in practice. However, in spite of impressive progress in the researches on the LCC, the most researches have only focused on the Deterministic or Probabilistic LCC analysis approach and general bridge at design stage. Thus, the goal of this study is to develop a practical and realistic methodology for the Life-Cycle Cost LCC-effective optimum decision-making based on reliability analysis of bridges at design stage. The proposed updated methodology is based on the concept of Life Cycle Performance(LCP) which is expressed as the sum of present value of expected direct/indirect maintenance costs with expected optimal maintenance scenario. The updated LCC methodology proposed in this study is applied to the optimum design problem of an actual highway bridge with Cable Stayed Bridges. In conclusion, based on the application of the proposed methods to an actual example bridge, it is demonstrated that a updated methodology for performance-based LCC analysis proposed in this thesis, shown applicably in practice as a efficient, practical, process LCC analysis method at design stage.

A Study on the Construction Cost Index for Calculating Conceptual Estimation : 1970-1999 (개략공사비 산출을 위한 공사비 지수 연구 : 1970-1999)

  • Nam, Song Hyun;Park, Hyung Keun
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.40 no.5
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    • pp.527-534
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    • 2020
  • A significant factor in construction work is cost. At early- and advanced-stage design, costs should be calculated to derive realistic cost estimates according to unit price calculation. Based on these estimates, the economic feasibility of construction work is assessed, and whether to proceed is determined. Through the Korea Institute of Civil Engineering and Building Technology, the construction cost index has been calculated by indirect methods after both the producer price index and construction market labor have been reprocessed to easily adjust the price changes of construction costs in Korea, and the Institute has announced it since 2004. As of January 2000, however, the construction cost index was released, and this has a time constraint on the correction and use of past construction cost data to the present moment. Variables were calculated to compute a rough construction cost that utilized past construction costs through surveys of the producer price index and the construction market labor force consisting of the construction cost index. After significant independent variables among the many variables were selected through correlation analysis, the construction cost index from 1970 to 1999 was calculated and presented through multiple regression analysis. This study therefore has prominent significance in terms of proposing a method of calculating rough construction costs that utilize construction costs that pre-date the 2000s.

Effect of Coexisting Ions on Electrokinetic Injection in Capillary Electrophoresis Analysis (모세관 전기영동 분석에서 계면 동전기 주입에 미치는 공존 이온의 영향)

  • Lee, Kwang-Woo;Jeon, Ji-Young;Lee, Kwang-Pill
    • Analytical Science and Technology
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    • v.9 no.1
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    • pp.35-42
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    • 1996
  • A rapid analytical method based on capillary electrophoresis is described for the determination of trace anions in high-purity chemicals which is used to prevent corrosion demage in nuclear power plants. Separations are carried out at 20kV using trimethylsilane-coated fused-silica capillary ($70cm{\times}50$ or $75{\mu}m$ i.d.) with the electrolyte of 5mM Chromate(pH=8). Detection was achieved using on-column indirect photometry at 254nm. The simultaneous analysis of inorganic anions, chloride, nitrate, sulfate, azide and phosphate was performed using methods of hydrodynamic(>1ppm) and / or electrokinetic(<1ppm) injection. The results of studies on the coexisting anions on analyte ions shows that peak responses of analyte in hydrodynamic injection is constant without effect of coexisting anions, but those of analysis in electrokinetic injection is strongly dependant upon the kind of coexisting anions and its ionic mobility. The analyte enrichment rate, hence peak response, is positive relationship with the resistance of the sample solution. Thus, appropriate measures, such as standard addition or internal standard technique, must be used to account for differences in conductance of standard and sample solutions.

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Characterization of canine coronaviruses isolated from the dogs with diarrhea in Korea (설사증 이환견(犬)으로 부터 분리(分離)한 canine coronavirus의 성상(性狀)에 관한 연구(硏究))

  • Lee, Byung-hyung;Jun, Moo-hyung;Park, Jong-hyeon;Hwang, Eui-kyung;Huh, Won
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.517-527
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    • 1994
  • An attempt was made to isolate a causative viral agents from the fecal specimens of the diseased dogs with the gastroenteritis symptoms. Two coronavirus-like agents were isolated by serial dilution end point method and plaque assay. The isolates were characterized in terms of cytopathology, antigenicity, replication, physicochemical and morphological properties. The results obtained through the experiment were as follows; 1. Among 7 fecal specimens collected from the dogs with enteric disease, 2(28.6%)coronavirus-like agents showing typical cytopathic effects of canine coronavirus were isolated, and designated as CCV D1 and CCV D2, respectively. 2. By the cross-neutralization test and indirect immunofluoresence antibody test, the isolates were antigenically indentified as the standard CCV. The viruses were replicated only in the cytoplasm of A-72 cells. 3. The isolates showed no haemagglutinating activity against the erythrocytes from 11 kinds of animals. 4. The electron microscopic observation for the isolates showed spherical and pleomorphic features, covered with club-shaped projections on the surface. The size of particles was ranged from 70 to 150nm. 5. In one-step growth curve for the isolates in A-72 cells, maximum titers of intracellular vius was $10^{4.6}$ $TCID_{50}/0.1ml$ at 46 hrs postinoculation(pi) of CCV Dl and $10^{4.4}$ $TCID_{50}/0.1ml$ at 34 hrs pi of CCV D2. The maximum titers of extracellular virus was $10^{5.5}$ $TCID_{50}/0.1ml$ at 58 hrs pi of CCV D1 and $10^{5.8}$ $TCID_{50}/0.1ml$ at 46 hrs pi of CCV D2. 6. In physicochemical property test, the isolates were very sensitive to choroform and were found to be RNA virus. The viruses was stable at pH 3.0 for 1 hr and at $22{^{\circ}C}$ for 5 hrs. However, infectivity titers reduced remarkably by treatment with $56{^{\circ}C}$ for 10min.

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NIR-TECHNOLOGY FOR RATIONALE SOIL ANALYSIS WITH IMPLICATIONS FOR PRECISION AGRICULTURE

  • Stenberg, Bo
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1061-1061
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    • 2001
  • The scope of precision agriculture is to reach the put up cultivation goals by adjusting inputs as precise as possible after what is required by the soil and crop potentials, on a high spatial resolution. Consequently, precision agriculture is also often called site specific agriculture. Regulation of field inputs “on the run” has been made possible by the GPS (Geographical Position System)-technology, which gives the farmer his exact real time positioning in the field. The general goal with precision agriculture is to apply inputs where they best fill their purpose. Thus, resources could be saved, and nutrient losses as well as the impact on the environment could be minimized without lowering total yields or putting product quality at risk. As already indicated the technology exists to regulate the input based on beforehand decisions. However, the real challenge is to provide a reliable basis for decision-making. To support high spatial resolution, extensive sampling and analysis is required for many soil and plant characteristics. The potential of the NIR-technology to provide rapid, low cost analyses with a minimum of sample preparation for a multitude of characteristics therefore constitutes a far to irresistible opportunity to be un-scrutinized. In our work we have concentrated on soil-analysis. The instrument we have used is a Bran Lubbe InfraAlyzer 500 (1300-2500 nm). Clay- and organic matter-contents are soil constituents with major implications for most properties and processes in the soil system. For these constituents we had a 3000-sample material provided. High performance models for the agricultural areas in Sweden have been constructed for clay-content, but a rather large reference material is required, probably due to the large variability of Swedish soils. By subdividing Sweden into six areas the total performance was improved. Unfortunately organic matter was not as easy to get at. Reliable models for larger areas could not be constructed. However, through keeping the mineral fraction of the soil at minimal variation good performance could be achieved locally. The influence of a highly variable mineral fraction is probably one of the reasons for the contradictory results found in the literature regarding organic matter content. Tentative studies have also been performed to elucidate the potential performance in contexts with direct operational implications: lime requirement and prediction of plant uptake of soil nitrogen. In both cases there is no definite reference method, but there are numerous indirect, or indicator, methods suggested. In our study, field experiments where used as references and NIR was compared with methods normally used in Sweden. The NIR-models performed equally or slightly better as the standard methods in both situations. However, whether this is good enough is open for evaluation.

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Mastitis Diagnostics by Near-infrared Spectra of Cows milk, Blood and Urine Using SIMCA Classification

  • Tsenkova, Roumiana;Atanassova, Stefka
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1247-1247
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    • 2001
  • Constituents of animal biofluids such as milk, blood and urine contain information specifically related to metabolic and health status of the ruminant animals. Some changes in composition of biofluids can be attributed to disease response of the animals. Mastitis is a major problem for the global dairy industry and causes substantial economic losses from decreasing milk production and reducing milk quality. The purpose of this study was to investigate potential of NIRS combined with multivariate analysis for cow's mastitis diagnosis based on NIR spectra of milk, blood and urine. A total of 112 bulk milk, urine and blood samples from 4 Holstein cows were analyzed. The milk samples were collected from morning milking. The urine samples were collected before morning milking and stored at -35$^{\circ}C$ until spectral analysis. The blood samples were collected before morning milking using a catheter inserted into the carotid vein. Heparin was added to blood samples to prevent coagulation. All milk samples were analyzed for somatic cell count (SCC). The SCC content in milk was used as indicator of mastitis and as quantitative parameter for respective urine and blood samples collected at same time. NIR spectra of blood and milk samples were obtained by InfraAlyzer 500 spectrophotometer, using a transflectance mode. NIR spectra of urine samples were obtained by NIR System 6500 spectrophotometer, using 1 mm sample thickness. All samples were divided into calibration set and test set. Class variable was assigned for each sample as follow: healthy (class 1) and mastitic (class 2), based on milk SCC content. SIMCA was implemented to create models of the respective classes based on NIR spectra of milk, blood or urine. For the calibration set of samples, SIMCA models (model for samples from healthy cows and model for samples from mastitic cows), correctly classified from 97.33 to 98.67% of milk samples, from 97.33 to 98.61% of urine samples and from 96.00 to 94.67% of blood samples. From samples in the test set, the percent of correctly classified samples varied from 70.27 to 89.19, depending mainly on spectral data pretreatment. The best results for all data sets were obtained when first derivative spectral data pretreatment was used. The incorrect classified samples were 5 from milk samples,5 and 4 from urine and blood samples, respectively. The analysis of changes in the loading of first PC factor for group of samples from healthy cows and group of samples from mastitic cows showed, that separation between classes was indirect and based on influence of mastitis on the milk, blood and urine components. Results from the present investigation showed that the changes that occur when a cow gets mastitis influence her milk, urine and blood spectra in a specific way. SIMCA allowed extraction of available spectral information from the milk, urine and blood spectra connected with mastitis. The obtained results could be used for development of a new method for mastitis detection.

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