• Title/Summary/Keyword: 96-well plate

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Cyclic behaviour of infilled steel frames with different beam-to-column connection types

  • Sakr, Mohammed A.;Eladly, Mohammed M.;Khalifa, Tarek;El-Khoriby, Saher
    • Steel and Composite Structures
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    • v.30 no.5
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    • pp.443-456
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    • 2019
  • Although numerous researchers demonstrated the significant difference in performance between the various beam-to-column connection types, most of the previous studies in the area of infilled steel frames focused on the behaviour of frames with welded connections. Therefore, there is a need for conducting studies on infilled steel frames with other common connection types (extended endplate with and without rib stiffeners, flush endplate and shear connections). In this paper, firstly, a two-dimensional finite-element model simulating the cyclic response of infilled steel frames was presented. The infill-frame interaction, as well as the interactions between connections' components, were properly modelled. Using the previously-validated model, a parametric study on infilled steel frames with five different beam-to-column connection types, under cyclic loading, was carried out. Several parameters, including infill material, fracture energy of masonry and infill thickness, were investigated. The results showed that the infilled frames with welded connections had the highest initial stiffness and load-carrying capacity. However, the infilled frames with extended endplate connections (without rib stiffeners) showed the greatest energy dissipation capacity and about 96% of the load-carrying capacity of frames with welded connections which indicates that this type of connection could have the best performance among the studied connection types. Finally, a simplified analytical model for estimating the stiffness and strength of infilled steel frames (with different beam-to-column connection types) subjected to lateral cyclic loading, was suggested.

A Study on the Shelf-life of Sausages in Refrigerated Storage (냉동 온도에서 소시지의 저장 수명에 관한 연구)

  • 이용욱;김종규
    • Journal of Food Hygiene and Safety
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    • v.10 no.2
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    • pp.111-131
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    • 1995
  • Quality changes of sausages in refrierated storage for 60 days were investigated. Nine types of sausages produced in Korea were stored at 4$^{\circ}C$(3~5$^{\circ}C$), and then chemical, microbiological, and sensory characteristics were evaluated on every 10 days. The proximate analysis showed considerable variation in fat (23.97%, 17.10~30.20%) with less variation in moisture (51.96%, 48.10~56.30%) and protein (12.96%, 11.40~13.95%), pH value decreased over time averaging from 6.31 to 6.22 with no significant difference. Water activity was consistent over refrigerated storage averaging 0.95. Volatile basic nitrogen (VBN) values were still within 20 mg% though VBN for all types significantly increased over time (p<0.05). Thiobarbituric acid (TBA) values were well below 1.0 though TBA showed significant differences among storage periods (p<0.05). Standard plate counts (SPCs) significantly increased during storage (p<0.05) while coliform group was not counted in all cases. SPCs reached 104 CFU/g in two types after 50 days' storage and were below 105 CFU/g in all types after 60 days' storage. Instrumental texture analysis showed that springiness, adhesiveness, and hardness signficantly decreased over time (p<0.05) while no change was observed in external color. significantly decreased over time the 60-day storage period (p<0.05) while cohesiveness, chewiness and gumminess did not change. Lightness, redness and yellowness of the internal Hunter color significantly decreased over time (p<0.05) while no change was observed in external color. Sensory profile showed that flavor, taste, texture and overall acceptable after 60 days' storage at 4$^{\circ}C$. However, the shelf-life of the samples should be decided in the consideration of the growth rate other spoilage flora coupled with the bacterial growth after 50 days' storage.

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The Effect of Hwangryunhaedoktang on Proliferations of Various Human Cancer Cells (황련해독탕이 수종의 인간 암세포 증식에 미치는 영향)

  • Sung, Hyun Kyung;Min, Sang Yeon;Kim, Jang Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.27 no.1
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    • pp.59-68
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    • 2013
  • Objectives The aim of this study is to investigate whether hwang-ryun-haedok-tang (HDT) affect proliferations of androgen-dependent LNCaP prostate cancer cells, androgen-independent PC-3, DU-145 prostate cancer cells, MCF-7 human breast cancer cells, A549, NCI-H292 human pulmonary cancer cells and K-562 human chronic myelogenous leukemia cells. Materials and Methods Effects of HDT on proliferations of each cancer cell line were investigated. 20,000 cells/well were plated in each well of 96-well culture plate. After 24 hrs, 0.01-10% of HDT in culture medium was added to cancer cells. The number of cells was counted by using SRB assay or direct cell counting method after 72 hours from drug treatment. Effect of baicalein or berebrine on proliferation was assessed according to the same method. Results (1) HDT inhibited proliferations of LNCaP, PC-3 and DU-145 prostate cancer cells. (2) HDT inhibited proliferation of MCF-7 breast cancer cells. (3) HDT also inhibited proliferations of A549, NCI-H292 pulmonary cancer cells and K-562 chronic myelogenous leukemia cells. (4) Baicalein and berberine also showed inhibitory effects on proliferations of prostate and breast cancer cells. Conclusion : HDT inhibited proliferations of human prostate, breast, pulmonary and blood cancer cells. These results suggest us the potential use of HDT as a chemopreventive or chemotherapeutic agent. Effect of HDT on human cancer should be further investigated using in vivo experimental models that can reflect pathophysiology of human cancer through another studies.

Ovarian cell aggregate culture in teleost, marine medaka (Oryzias dancena): basic culture conditions and characterization

  • Jae Hoon, Choi;Seung Pyo Gong
    • Journal of Animal Reproduction and Biotechnology
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    • v.39 no.1
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    • pp.19-30
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    • 2024
  • Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

CYTOTOXICITY OF DENTIN BONDING SYSTEM ON L929 CELLS (상아질 결합제의 L929 세포 독성에 관한 연구)

  • Im, Mi-Kyung;Ji, Jung-Ho;Kim, Sang-Seop
    • Restorative Dentistry and Endodontics
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    • v.22 no.1
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    • pp.209-219
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    • 1997
  • The research of the dentin bonding system was mainly on the chemistry and bonding strength. And in vitro assessement of biocompatibility of dentin bonding system was not completely developed. The purpose of this study was to evaluate the cytotoxic effect of several dentin primers. Scotchbond Multi-Purpose (3M Dental Products. USA). Gluma (BayerDental. Germany). All-Bond (Bisco. USA). ProBond (CaulkDensply, USA) and VeridonFil (Dongyang Nylon. Korea) were included. Cytotoxicity was tested using MTT cell viability test. 0.5 ul. 1 ul. 2 ul and 10 ul of each primer were added to the 96 well plate of incubated L929 cell lines. After 30-minute. 1. 4. 24 and 72-hour exposures. absorbance of L929 cells was observed with ELISA reader. All data were analyzed using t-test. All primers showed cytotoxicity on L929 cells under every conditions used in this study. Absorbance of L929 cells was decreased by time. Scotch bond group exhibited the lowest absorbance value in all exposure time and value.

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Screening of New Antibiotics Inhibiting Bacterial Peptide Deformylase (PDF) (세균의 Peptide Deformylase(PDF)를 억제하는 새로운 항균물질의 스크리닝)

  • 곽진환;김현주;설민정;서병선;이종국;최수영
    • YAKHAK HOEJI
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    • v.47 no.3
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    • pp.184-189
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    • 2003
  • Peptide deformylase (PDF) is essential and unique to bacteria, thus making it an attractive target for the discovery of novel antibacterial drugs. PDF deformylates the N-formylmethionine of newly synthesized polypeptides in prokaryotes. In this study, a pdf gene from Staphylococcus aureus 6538p was cloned in pET-14b vector and PDF protein was over-produced in Escherichia coli BL21 (DE3). NH$_2$-terminal His-tagged PDF protein was purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography. Enzymatic activity of purified 6xHis-tagged PDF was tested on the substrate (formyl-Methionine-Alanine-Serine) by formate dehydrogenase-coupled spectrometric assay of peptide deformylase. For the discovery of new PDF inhibitors from chemical libraries and culture broths of soil bacteria, a target-oriented screening system using a 96-well plate was developed. About 3,000 commercial chemical libraries were tested in this screening system, and 2 chemicals (0.07%) among them showed an inhibitory activity against PDF enzyme. This result showed that a new screening system can be used for the discovery of new PDF inhibitors.

Screening for Various Herb Medicines Extracts against Human immunodeficiency virus & Herpes simplex virus type I and Herpes simplex virus type II (바이러스 질환 필요처방의 선정을 위한 수종 한방 처방의 효능연구)

  • Lim, Seong-Woo;Ahn, Duk-Kyun;Woo, Eun-Rhan;Park, Ho-Koon
    • The Journal of Korean Medicine
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    • v.19 no.2
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    • pp.326-336
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    • 1998
  • 면역 결핍 바이러스와 허피즈 바이러스 -1,2에 대한 한약의 항바이러스 작용을 관찰하기 위하여, 세포 바이러스 검색에 초기 독성과 감염 바이러스의 생존 세포 수를 비교적 단기간내 볼 수 있는 96-well plate를 이용한 MTT assay로 측정하였다. 본 실험은 예비 실험 단계에서 사용된 총 85가지의 한약중 단미제 6가지와 복합처방 44가지를 선정하였으며 복합처방은 유사한 치법으로 분류하였고 한약의 시료 추출은 순수하게 물로 전탕하여 여과하였다. 실험 결과 파두와 맥아가 면액 결핍 바이러스에 대해 감염초기 유의성이 있었으며 호장근, 갈근우방자탕과 형방패독산이 허피즈 바이러스-1,2에 대해 감염초기 유의성이 있었으나 실험의 시간 경과에 따른 지속적인 약효 안정성을 보이지는 못하였다. 이 실험을 통하여 한약을 이용한 우수한 바이러스 치료를 개발하기 위해서는 단미제나 복합 처방에서 항바이러스 작용이 큰 유효성분의 대량 분리와 세포 실험에 있어서 오차를 줄 일 수 있는 세포면역학적 실험의 도입 등이 필요할 것으로 사료된다.

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A MALDI-MS-based Glucan Hydrolase Assay Method for Whole-cell Biocatalysis

  • Ahn, Da-Hee;Park, Han-Gyu;Song, Won-Suk;Kim, Seong-Min;Jo, Sung-Hyun;Yang, Yung-Hun;Kim, Yun-Gon
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.69-77
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    • 2019
  • Screening microorganisms that can produce glucan hydrolases for industrial, environmental, and biomedical applications is important. Herein, we describe a novel approach to perform glucan hydrolase screening-based on analysis of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) spectra-which involves degradation of the oligo- and polysaccharides. As a proof-of-concept study, glucan hydrolases that could break down glucans made of several glucose units were used to demonstrate the MALDI-MS-based enzyme assay. First, the enzyme activities of ${\alpha}$-amylase and cellulase on a mixture of glucan oligosaccharides were successfully discriminated, where changes of the MALDI-MS profiles directly reflected the glucan hydrolase activities. Next, we validated that this MALDI-MS-based enzyme assay could be applied to glucan polysaccharides (i.e., pullulan, lichenan, and schizophyllan). Finally, the bacterial glucan hydrolase activities were screened on 96-well plate-based platforms, using cell lysates or samples of secreted enzyme. Our results demonstrated that the MALDI-MS-based enzyme assay system would be useful for investigating bacterial glucoside hydrolases in a high-throughput manner.

Antibiofilm Activity of a Curcuma zedoaria Rosc Rhizome Extract against Methicillin-Resistant and Susceptible Staphylococcus aureus

  • Tabunhan, Sompong;Tungsukruthai, Parunkul
    • Microbiology and Biotechnology Letters
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    • v.50 no.2
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    • pp.193-201
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    • 2022
  • Methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) are major causes of hospital- and community-acquired infections. The treatment of biofilm-related infections caused by these bacteria is a global healthcare challenge. Therefore, the development of alternative therapeutics is required. An essential oil extracted from Curcuma zedoaria (CZ) Rosc, also known as white turmeric, has been reported to possess various antimicrobial activities. In the present study, we evaluated the antibiofilm activities of an ethanolic extract of the CZ rhizome against MRSA and MSSA. The results showed that the CZ extract with the highest sub-minimum inhibitory concentration (sub-MIC), 1/2 MIC (0.312 mg/ml), significantly inhibited biofilm production by up to 80-90% in both tested strains. Subsequently, we evaluated the ability of the CZ extract to prevent cell-surface attachment to a 96-well plate and extracellular DNA (eDNA) release from the biofilm. The CZ extract demonstrated an inhibitory effect on bacterial attachment and eDNA release from the biofilm biomass. The CZ extract may inhibit biofilm formation by preventing eDNA release and cell-surface attachment. Therefore, this CZ extract is a potential candidate for the development of alternative treatments for biofilm-associated MRSA and MSSA infections.

Characteristics of Chungkookjang Produced by Bacillus subtillus MC31 (B. subtilis MC 31를 이용한 청국장의 품질특성)

  • Mann, So-Yon;Kim, Eun-Ah;Lee, Ga-Young;Kim, Ro-Ui;Hwang, Dae-Youn;Son, Hong-Joo;Lee, Byong-Won;Lee, Chung-Yeol;Kim, Dong-Seob
    • Journal of Life Science
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    • v.23 no.4
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    • pp.560-568
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    • 2013
  • Chungkookjang was fermented by B. subtilis MC31, a ${\gamma}$-amino butyric acid (GABA) producing microorganism. The characteristics of Chungkookjang were investigated while fermenting. Twenty four amino acids were detected in Chungkookjang, leucine was the highest of them all. Total cell populations of B. subtilis MC31 phase were between log $9.52{\pm}0.5$ ~ log $9.049{\pm}0.5$ CFU/g at stationary phase. Contents of moisture, crude ash, crude protein, crude lipid and crude fiber are $61.07{\pm}0.01%$, $1.52{\pm}0.01%$, $17.66{\pm}0.04%$, $8.96{\pm}0.03%$ and 2.61%, respectively. Contents of ammonia type nitrogen, amino type nitrogen and reducing sugar were increased during fermentation at $40^{\circ}C$ for 72 hr, however those of titratable acidity and total sugar were decreased. pH was slowly alkalized during fermentation. Viscous substance and protease contents in Chungkookjang were $4.7{\pm}0.05%$ and $0.519{\pm}7.36$ g/l, apiece. When the fibrin plate and Robbin method for fibrinolytic activity were applied, B. subtilis MC31 showed high activity. These results suggested that B. subtilis MC31 is suitable to be used as a starter to enhance the quality of Chungkookjang.