• Title/Summary/Keyword: 96-well plate

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Buckling Analysis of Sandwich Plate with Thin Faces (얇은 면재를 갖는 샌드위치 평판의 좌굴강도 해석)

  • H.K. Yang;J.W. Lee;Y.H. Kim
    • Journal of the Society of Naval Architects of Korea
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    • v.32 no.4
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    • pp.87-96
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    • 1995
  • This study is concerned with the overall buckling analysis of sandwich plates under biaxial loads by applying the Rayleigh-Ritz method, which are considered to buckle simultaneously in overall from of core and thin faces together. In order to study the effects of boundary conditions on the buckling behaviors, the simply supported, flexed and it's combined boundary conditions are considered as well as the effects of material characteristics of core and thin faces of sandwich plates on the buckling behaviors.

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High-throughput SNP Genotyping by Melting Curve Analysis for Resistance to Southern Root-knot Nematode and Frogeye Leaf Spot in Soybean

  • Ha, Bo-Keun;Boerma, H. Roger
    • Journal of Crop Science and Biotechnology
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    • v.11 no.2
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    • pp.91-100
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    • 2008
  • Melting curve analysis of fluorescently labeled DNA fragments is used extensively for genotyping single nucleotide polymorphism(SNP). Here, we evaluated a SNP genotyping method by melting curve analysis with the two probe chemistries in a 384-well plate format on a Roche LightCycler 480. The HybProbe chemistry is based on the fluorescence resonance energy transfer(FRET) and the SimpleProbe chemistry uses a terminal self-quenching fluorophore. We evaluated FRET HybProbes and SimpleProbes for two SNP sites closely linked to two quantitative trait loci(QTL) for southern root-knot nematode resistance. These probes were used to genotype the two parents and 94 $F_2$ plants from the cross of PI 96354$\times$Bossier. The SNP genotypes of all samples determined by the LightCycler software agreed with previously determined SSR genotypes and the SNP genotypes determined on a Luminex 100 flow cytometry instrument. Multiplexed HybProbes for the two SNPs showed a 98.4% success rate and 100% concordance between repeats two of the same 96 DNA samples. Also, we developed a HybProbe assay for the Rcs3 gene conditioning broad resistance to the frogeye leaf spot(FLS) disease. The LightCycler 480 provides rapid PCR on 384-well plate and allows simultaneous amplification and analysis in approximately 2 hours without any additional steps after amplification. This allowed for a reduction of the potential contamination of PCR products, simplicity, and enablement of a streamlined workflow. The melting curve analysis on the LightCycler 480 provided high-throughput and rapid SNP genotyping and appears highly effective for marker-assisted selection in soybean.

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Safety assessment of generation III nuclear power plant buildings subjected to commercial aircraft crash part III: Engine missile impacting SC plate

  • Xu, Z.Y.;Wu, H.;Liu, X.;Qu, Y.G.;Li, Z.C.;Fang, Q.
    • Nuclear Engineering and Technology
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    • v.52 no.2
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    • pp.417-428
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    • 2020
  • Investigations of the commercial aircraft impact effect on nuclear island infrastructures have been drawing extensive attention, and this paper aims to perform the safety assessment of Generation III nuclear power plant (NPP) buildings subjected to typical commercial aircrafts crash. At present Part III, the local damage of the rigid components of aircraft, e.g., engine and landing gear, impacting the steel concrete (SC) structures of NPP containment is mainly discussed. Two typical SC target panels with the thicknesses of 40 mm and 100 mm, as well as the steel cylindrical projectile with a mass of 2.15 kg and a diameter of 80 mm are fabricated. By using a large-caliber air gas gun, both the projectile penetration and perforation test are conducted, in which the striking velocities were ranged from 96 m/s to 157 m/s. The bulging velocity and the maximal deflection of rear steel plate, as well as penetration depth of projectile are derived, and the local deformation and failure modes of SC panels are assessed experimentally. Then, the commercial finite element program LS-DYNA is utilized to perform the numerical simulations, by comparisons with the experimental and simulated projectile impact process and SC panel damage, the numerical algorithm, constitutive models and the corresponding parameters are verified. The present work can provide helpful references for the evaluation of the local impact resistance of NPP buildings against the aircraft engine.

High-Throughput DNA Extraction Method for Marker Analysis in Rice Grain (대량의 쌀 시료 분석을 위한 DNA 추출법)

  • Choi, Young-Deok;Lee, Hae-Kwang;Lee, Yun-Suk;Yun, Jeong-Hee;Kim, Su-Jeong;Park, Seong-Whan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.269-273
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    • 2006
  • The study of molecular markers to improve crops largely depends on the availability of rapid and of efficient DNA extraction methods. Here we developed a cheap and convenient method to isolate genomic DNA from rice grains suitable for large-scale microsatellite analysis. We confirmed that the isolated rice DNA is suitable for PCR analysis with STS marker and SNP marker, as well as microsatellite marker. Further, we established high-throughput DNA extraction system in a 96-well plate format which make it possible high-throughput analysis of microsatellite markers with rice grains. This implies that the new method could be a useful tool for other types of marker analysis in large scale.

EFFECTS OF ALENDRONATE AND PAMIDRONATE ON THE PROLIFERATION AND THE ALKALINE PHOSPHATASE ACTIVITY OF HUMAN BONE MARROW DERIVED MESENCHYMAL STEM CELLS (Alendronate와 Pamidronate가 인간 골수유래 간엽줄기세포의 증식과 알칼리성 인산분해효소 활성에 미치는 영향)

  • Kim, Young-Ran;Ryu, Dong-Mok;Kwon, Yong-Dae;Yun, Yong-Pil
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.35 no.6
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    • pp.397-402
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    • 2009
  • The purpose of this study is to investigate the effects of alendronate and pamidronate on proliferation and the alkaline phosphatase activity of human bone marrow derived mesenchymal stem cells and to relate the results with bisphosphonate related osteonecrosis of the jaw(BRONJ). With the consent of patients with no systemic disease and undergoing iliac bone graft, cancellous bone was collected to obtain human bone marrow derived mesenchymal stem cells through cell culture. 96 well plate were prepared with a concentration of $10^4$cell/ well. Alendronate and pamidronate were added to each well with the concentration of $10^{-6}M$, $10^{-8}M$ and $10^{-10}M$, respectively. Then proliferation capacity of each well was evaluated with the cell counting kit. 24 well plates were prepared with a concentration of $10^5$cell/ml/well and with the bone supplement, alendronate and pamidronate were added with the concentration of $10^{-6}M$, $10^{-8}M$ and $10^{-10}M$, respectively on each plate. The plates were cultured for either 24 or 72 hours. Then the cells were sonicated to measure the alkaline phosphatase activity and protein assay was done to standardize the data for analysis. As the concentration of alendronate or pamidronate added to the culture increased, the proliferation capacity of the cells decreased. However, no statistical significance was found between the group with $10^{-10}M$ of bisphophonate and the control group. Pamidronate was not capable of increasing the alkaline phosphatase activity in all trials. However, alkaline phosphatase activity increased with 24 hours of $10^{-8}M$ of alendronate treatment and with 48 hours of $10^{-10}M$ of alendronate treatment. Cell toxicity increased as the bisphosphonate concentration increased. This seems to be associated with the long half life of bisphosphonate, resulting in high concentration of bisphosphonate in the jaw and thus displaying delayed healing after surgical procedures. Alendronate has shown to increase the alkaline phophatase activity of human bone marrow derived mesenchymal stem cells. However, this data is insufficient to conclude that alendronate facilitates the differentiation of human bone marrow derived mesenchymal stem cells. Further studies on DNA level and animal studies are required to support these results.

CYTOTOXIC EFFECTS OF SOLUBLE EXTRACTS FROM RERIODONTAL DRESSINGS ON HUMAN GINGIVAL FIBROBLASTS (치은 섬유아세포에 대한 치주포대 추출물의 세포동성에 관한 연구)

  • Yang, Seung-Han;Lee, Man-Sup;Park, Joo-Bong
    • Journal of Periodontal and Implant Science
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    • v.24 no.3
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    • pp.661-670
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    • 1994
  • It is well known that the application of dressings after periodontal surgery have benefits to provide the comforts to patient and to promote the healing process with action of bleeding control and temporary stabilization for the operated mobile teeth. But until recently the relationship between periodontal dressings and cells which are composed of periodontium has not been clear. The purpose of this study was to evaluate the cytotoxic effect of soluble extracts from the four different kinds of periodontal dressings, two of them were eugenol type (K.H.pack, Wondrpak) and the others were non-eugenol type (Coe-pak, Periocare), on the human gingival fibroblasts in vitro. Human gingival fibroblasts were primarily cultured from gingiva around third molar during the extraction for preventive purposes. Extracts solution were prepared with culture medium by means of imersing the consistent size of periodontal dressing made from plastic mold. Cell were inoculated into the 24 well plate with $3\;{\times}\;10^4\;cells/well$ of medium at $37\;^{\circ}C$, 100% of humidity, 5% of $CO_2$, incubator for 24 hours. After discard of the supernatant of medium, those cells were cultured with original, 1/2, 1/5, 1/10 diluted soluble extract for 24, 48 and 72 hours, and counted the number of cells using the hemocytometer at each designed time and concentration. Also, the cytotoxic effect of soluble extract was measured by Wataha's MTT assay method. In briefly, cells were inoculated and cultured into 96 well culture plate with $2\;{\times}\;10^4\;cells/well$ for 24 hours. Soluble extracts were applied to cultured cells and incubated for 48 hours at same condition. $50\;{\mu}l$ of MTT solution and DMSO were added into each well for the detection of absorbance with ELISA reader. The measured data were calculated by value of colorimetric assay for survival rate. The results were as follows ; In the case of eugenol type of dressing, original, 1/2 and 1/5 diluted extracts of K.H.pack showed very low survival rate. And original extract of Wondrpak showed strong cytotoxic effect and 1/2 diluted extract showed moderate cytotoxic effect. In the case of Non-eugenol type of dressings, only original extract of Coe-pak revealed strong cytotoxic effect and Periocare had little cytotoxic effect. It is concluded that eugenol type of dressings showed more cytotoxic effect than non-eugenol types. This study suggest that use of non-eugenol dressings after periodontal surgery is recommended.

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Controlling Effect of Some Environmentally Friendly Agents on Garlic Leaf Blight in Garlic (마늘 잎마름병에 대한 친환경제제 방제효과)

  • Ryu, Young-Hyun;Huh, Chang-Seok;Kim, Dong-Geun;Yeon, Il-Kwon;Jo, Woo-Sik;Ryu, Jung-A
    • Korean Journal of Organic Agriculture
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    • v.23 no.2
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    • pp.347-357
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    • 2015
  • This experiment was conducted to test the efficiency of environmentally friendly materials for controlling garlic leaf blight by Stemphylium botryosum. Fifty five kinds of environmental friendly control agents are obtained from commercial market and are tested for spore germination using 96 well plate and among them, five agents (copper-, sulfur-, medinal herb extract-, sulfur+sodium bicarbonate- and oligo chitosan based compound) are selected for field test from 2012 and 2013 year. With reference of 2012 year test result, copper and oligo chitosan based compounds are chosen as 2013 year test. When the first symptoms were appeared early May season and environmental friendly control agents are applied as prompt as possible, the control value of copper and oligo chitosan based agents are 54% and 90% respectively as compared to the occasion of chemical agent Antracol WP (propineb 70%) and yield of bulbs are increased by 16% and 34% against untreated control and marketable garlic bulb yield were 79% and 95% against Antracol WP treatment, respectively. From this result, oligo chitosan based compound can be a good organic control agent candidate for garlic leaf blight disease in organic garlic cultivation.

Spectrophotometric microtitre assay for rapid screening of membrane-disrupting herbicides (분광분석을 이용한 막과산화작용 제초제의 신속한 검정법)

  • Kwon, Ok-Kyung;Cho, Kwang-Yun;Kim, Jin-Seog
    • The Korean Journal of Pesticide Science
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    • v.4 no.1
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    • pp.11-18
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    • 2000
  • This study was conducted to establish an efficient screening method of membrane-disrupting herbicides, based on the result that membrane leakage leading to evolution of carbonyl compounds and aldehydes could be colorimetrically assessed by formation of stable chromophores with MBTH and ferric chloride. Assay conditions including plant material, light intensity, irradiation time, concentrations of reagents for colorization, and reaction time were optimized, and activities of different types of known herbicides were investigated with cucumber cotyledon discs. That result indicated that this assay was sensitive to membrane-disrupting herbicides, such as protoporphyrinogen oxidase inhibitors (acifluorfen, oxyfluorfen, oxadiazon) and an electron interceptor from photosystem I(paraquat). With seven compounds newly synthesized, highly positive correlation was observed between herbicidal activities measured by greenhouse bioassay and MBTH method. Because only a single disc(4 mm diameter) from cucumber cotyledon in 60 ${\mu}L$ of incubation solution was required to assay, a large number of herbicides could be tested simultaneously using 96-well microtitre plates. But the assay efficiency is not likely to significantly increase because of the boiling step during the reaction.

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Full validation of high-throughput bioanalytical method for the new drug in plasma by LC-MS/MS and its applicability to toxicokinetic analysis

  • Han, Sang-Beom
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2006.11a
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    • pp.65-74
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    • 2006
  • Modem drug discovery requires rapid pharmacokinetic evaluation of chemically diverse compounds for early candidate selection. This demands the development of analytical methods that offer high-throughput of samples. Naturally, liquid chromatography / tandem mass spectrometry (LC-MS/MS) is choice of the analytical method because of its superior sensitivity and selectivity. As a result of the short analysis time(typically 3-5min) by LC-MS/MS, sample preparation has become the rate- determining step in the whole analytical cycle. Consequently tremendous efforts are being made to speed up and automate this step. In a typical automated 96-well SPE(solid-phase extraction) procedure, plasma samples are transferred to the 96-well SPE plate, internal standard and aqueous buffer solutions are added and then vacuum is applied using the robotic liquid handling system. It takes only 20-90 min to process 96 samples by automated SPE and the analyst is physically occupied for only approximately 10 min. Recently, the ultra-high flow rate liquid chromatography (turbulent-flow chromatography)has sparked a huge interest for rapid and direct quantitation of drugs in plasma. There is no sample preparation except for sample aliquotting, internal standard addition and centrifugation. This type of analysis is achieved by using a small diameter column with a large particle size(30-5O ${\mu}$m) and a high flow rate, typically between 3-5 ml/min. Silica-based monolithic HPLC columns contain a novel chromatographic support in which the traditional particulate packing has been replaced with a single, continuous network (monolith) of pcrous silica. The main advantage of such a network is decreased backpressure due to macropores (2 ${\mu}$m) throughout the network. This allows high flow rates, and hence fast analyses that are unattainable with traditional particulate columns. The reduction of particle diameter in HPLC results in increased column efficiency. use of small particles (<2 urn), however, requires p.essu.es beyond the traditional 6,000 psi of conventional pumping devices. Instrumental development in recent years has resulted in pumping devices capable of handling the requirements of columns packed with small particles. The staggered parallel HPLC system consists of four fully independent binary HPLC pumps, a modified auto sampler, and a series of switching and selector valves all controlled by a single computer program. The system improves sample throughput without sacrificing chromatographic separation or data quality. Sample throughput can be increased nearly four-fold without requiring significant changes in current analytical procedures. The process of Bioanalytical Method Validation is required by the FDA to assess and verify the performance of a chronlatographic method prior to its application in sample analysis. The validation should address the selectivity, linearity, accuracy, precision and stability of the method. This presentation will provide all overview of the work required to accomplish a full validation and show how a chromatographic method is suitable for toxirokinetic sample analysis. A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method developed to quantitate drug levels in dog plasma will be used as an example of tile process.

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Improvement of Wear Resistance of Aluminum by Metal-Ceramic Particle Composite Layer (알루미늄표면에 금속-세라믹입자 복합첨가에 의한 내마모성개선)

  • ;;;中田一博;松田福久
    • Journal of Welding and Joining
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    • v.15 no.6
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    • pp.96-104
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    • 1997
  • The present study was aimed to correlate the microstructure and the hardness as well as the wear resistance of the metal-ceramic particulated composite layer on the pure Al plate. The composite layers were constructed by the addition of TiC particles on the surface of Al-Cu alloyed layers by PTA overlaying process. Initially, the Al-Cu alloyed layers were achieved by the deposition of Al-(25 ~ 48%) Cu alloys on the pure Al plate by TIG process. It was revealed that TiC particles were uniformly dispersed without any reaction with matrix in the composite layer. The volume fraction of TiC particles (TiC V F) increased from 12% to 55% with increasing the number of pass of composite layer. Hardnesses of (Al-48%Cu + TiC (3&4layers)) composite layer were Hv450 and Hv560, respectively, due to the increase of TiC V/F. Hardnesses of (Al-Cu + TiC) composite layers decreased gradually with insreasing temperature from 100$^{\circ}$C to 400$^{\circ}$C, and hardnesses at 400$^{\circ}$C were then reached to 1/5 - 1/10 of room temperature hardness depending on the construction of composite layers. The Specific wear of (Al + Tic) layer and Al-48%Cu alloyed layer decreased to 1/10 of the of pure Al, while the specific wear of (Al-48%Cu + TiC (4 layers)) composite layer exhibited 1/15 of that of steel such as SS400 and STS304.

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