• Title/Summary/Keyword: Cell Extraction

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Graded Noise Elimination and Cluster Boundary Extraction in Confocal Sliced Images (공초점 단층 이미지에서 수준별 잡음제거와 클러스터 경계선 추출)

  • Cho, Mi-Gyung;Kim, Jin-Seok;Shim, Jae-Sool
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.15 no.12
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    • pp.2697-2704
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    • 2011
  • In tissue engineering area, researchers observe symbiotic relationship such as proliferation, interaction, division apoptosis with time between cells in process of the 3D cell culture in hydrogels. The 3D cell culture process can be taken photographs into sliced images using confocal microscope. Symbiotic mechanism and changes of cell behaviors can be observed and analyzed from the images acquired by confocal microscope. In this paper, we proposed and developed graded noise elimination method and cluster boundary extraction method to extract boundaries information from sliced confocal images acquired in process of the 3D cell culture in hydrogels. The experiment based algorithm showed excellent performance for eliminating noises that have very small millet-shaped size. It is also showed to extract exact boundaries information for even complex clusters.

Electron Microscopy of Cell Walls of Saccharomces cervisiae and Mycobacterium phlei in the process of DNA extraction (Saccharomyces cerevisiae와 Mycobacterium phlei에서 DNA유출에 따른 세포벽의 전자현미경적 고찰)

  • Lee, Kil-Soo;Cho, She-Hoon;Kim, Woon-Soo;Lew, Joon
    • Korean Journal of Microbiology
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    • v.13 no.3
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    • pp.109-115
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    • 1975
  • DNA's were extracted from Saccharomyces cerevisiae and Mycobacterium phlei and the damaging cell walls of these microoragnisms were examined under an electron microscope in the extraction process in which a number of physico-chemical tratments of cells was involved. While the DNA was easily extracted from S. cerevisiae using conventional meylelded very little DNA, of M. phlei was extremely difficult to isolate and yielded very little DNA, applying various methods of isolation published earlier. When the cell walls of S. cerevisiae were examined with the electron microscope, they were not yet damaged even after the cells were treated with sodium lauryl sulfate(SLS) and ethylene diamine tetracetic acid(EDTA), but they were completely destroyed by the treatment of sodium perchlorate followed by the addition of chloroform and a vigorous agitation. Oozing cytoplasm through the broken cell walls was also observed. In the extraction of DNA from M.phlei, the pronase was not effective at the aerobic environment of the sample. When phenol was applied at the last step of DNA isolation, an extreme damage mass yielding little DNA into the solution. Unlike the cells of S.cerevisiae.M.phlei cells showed a tendency of aggregation, thus the destruction of cell walls by sodium hydroxide was seen only on the walls of peripheral cells in the aggregated mass, leaving the walls of the inner cells undamaged.

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Identification of bioactive components behind the antimicrobial activity of cow urine by peptide and metabolite profiling

  • Rohit Kumar;Jai Kumar Kaushik;Ashok Kumar Mohanty;Sudarshan Kumar
    • Animal Bioscience
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    • v.36 no.7
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    • pp.1130-1142
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    • 2023
  • Objective: Cow urine possesses several bioactive properties but the responsible components behind these bioactivities are still far from identified. In our study, we tried to identify the possible components behind the antimicrobial activity of cow urine by exploring the peptidome and metabolome. Methods: We extracted peptides from the urine of Sahiwal cows belonging to three different physiological states viz heifer, lactation, and pregnant, each group consisting of 10 different animals. The peptides were extracted using the solid phase extraction technique followed by further extraction using ethyl acetate. The antimicrobial activity of the aqueous extract was evaluated against different pathogenic strains like Staphylococcus aureus, Escherichia coli, and Streptococcus agalactiae. The safety of urinary aqueous extract was evaluated by hemolysis and cytotoxicity assay on the BuMEC cell line. The urinary peptides were further fractionated using high-performance liquid chromatography (HPLC) to identify the fraction(s) containing the antimicrobial activity. The HPLC fractions and ethyl acetate extract were analyzed using nLC-MS/MS for the identification of the peptides and metabolites. Results: A total of three fractions were identified with antimicrobial activity, and nLC-MS/MS analysis of fractions resulted in the identification of 511 sequences. While 46 compounds were identified in the metabolite profiling of organic extract. The urinary aqueous extract showed significant activity against E. coli as compared to S. aureus and S. agalactiae and was relatively safe against mammalian cells. Conclusion: The antimicrobial activity of cow urine is a consequence of the feeding habit. The metabolites of plant origin with several bioactivities are eliminated through urine and are responsible for their antimicrobial nature. Secondly, the plethora of peptides generated from the activity of endogenous proteases on protein shed from different parts of tissues also find their way to urine. Some of these sequences possess antimicrobial activity due to their amino acid composition.

Effects of $Cinnamomi$ $Cortex$ Pharmacopuncture on LPS-induced Inflammatory Response in Macrophage (육계약침액이 LPS로 유도된 대식세포의 염증반응에 미치는 영향)

  • Kim, Hyeong-Seok;Roh, Jeong-Du
    • Journal of Acupuncture Research
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    • v.29 no.1
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    • pp.15-24
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    • 2012
  • Objectives : In recent years, many studies have been widely researching anti-inflammation effect of various medicinal plants. $Cinnamomi$ $Cortex$ was not enough in researching of the anti-inflammation. Moreover, there is no comparative study about extraction methods. Therefore, we investigated the inhibitory effects of $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction on Nitric oxide(NO), Prostaglandin E2(PGE2) production, Cyclooxygenase(COX)-2, inducible NOS(iNOS) expression and extracellular signal regulate kinase(ERK)1/2 phosphorylation in lipopolysaccharide(LPS) induced RAW 264.7 macrophage cell. Methods : $Cinnamomi$ $Cortex$ was extracted by EtOH and Hot water. RAW 264.7 macrophage cell viability was measured by MTT assay. Effect of $Cinnamomi$ $Cortex$ pharmacopuncture on NO and PGE2 production in LPS induced macrophages was accessed by Griess assay and enzyme-linked immunospecific assay(ELISA), respectively. Inhibition effect on COX-2, iNOS expression and ERK1/2 phosphorylation was examined by Immunoblotting assay. Results : 1. Cytotoxic effect of $Cinnamomi$ $Cortex$ pharmacopuncture by Hot water extraction in RAW 264.7 macrophages was not appeared, except $3125{\mu}g/m{\ell}$. And cytotoxic effect was not appeared in EtOH extraction method. 2. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited NO production in LPS induced macrophages significantly. 3. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited PGE2 production in LPS induced macrophages significantly. 4. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited COX-2, iNOS expression in LPS induced macrophages. Especially, it has been confirmed that COX-2, iNOS expression were effectively inhibited in Hot water extraction. 5. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited ERK1/2 phosphorylation in LPS induced macrophages. Especially, it has been confirmed that ERK1/2 phosphorylation was effectively inhibited in Hot water extraction. Conclusions : According to the results, $Cinnamomi$ $Cortex$ pharmacopuncture suppresses NO, PGE2 production, COX-2, iNOS expression and ERK1/2 phosphorylation in LPS induced macrophages. It has a potential for treating various inflammatory diseases, and Hot water extraction method could be used more extensively than EtOH extraction method.

Cytotoxic Effects of Extracts from Tremella fuciformis Strain FB001 on the Human Colon Adenocarcinoma Cell Line DLD-l

  • Kim, Kyung-Ai;Chang, Hyun-You;Choi, Sung-Woo;Yoon, Jeong-Weon;Lee, Chan
    • Food Science and Biotechnology
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    • v.15 no.6
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    • pp.889-895
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    • 2006
  • Cytotoxic effects of extracts from Tremella fuciformis strain FB001 were evaluated on the DLD-1 human colon adenocarcinoma cell line and the content of polyphenolic compounds in the extracts were analyzed. Hexane, chloroform, and ethyl acetate subfractions (experimental setting I) exhibited cytotoxic effects on the human colon adenocarcinoma DLD-1 cell line with $IC_{50}$ values of 350, 400, and 450 ppm, respectively. When T. fuciformis was extracted sequentially with ether, ethyl acetate, chloroform, and ethanol (experimental setting II), the ether extract demonstrated potent cytotoxicity with an $IC_{50}$ value of 150 ppm, followed by ethyl acetate and chloroform fractions. If the first extraction solvent was chloroform instead of ether (experimental setting III), exposure of the cell line to chloroform, ethyl acetate, and ether extracts at 1,000 ppm led to cell death. High levels of phenolic compounds were estimated for all hydrophobic extracts, which exhibited cytotoxic effects. We propose that this useful information gives additional support to our understanding of the biology and utility of this particular mushroom.

A Prediction of Chip Quality using OPTICS (Ordering Points to Identify the Clustering Structure)-based Feature Extraction at the Cell Level (셀 레벨에서의 OPTICS 기반 특질 추출을 이용한 칩 품질 예측)

  • Kim, Ki Hyun;Baek, Jun Geol
    • Journal of Korean Institute of Industrial Engineers
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    • v.40 no.3
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    • pp.257-266
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    • 2014
  • The semiconductor manufacturing industry is managed by a number of parameters from the FAB which is the initial step of production to package test which is the final step of production. Various methods for prediction for the quality and yield are required to reduce the production costs caused by a complicated manufacturing process. In order to increase the accuracy of quality prediction, we have to extract the significant features from the large amount of data. In this study, we propose the method for extracting feature from the cell level data of probe test process using OPTICS which is one of the density-based clustering to improve the prediction accuracy of the quality of the assembled chips that will be placed in a package test. Two features extracted by using OPTICS are used as input variables of quality prediction model because of having position information of the cell defect. The package test progress for chips classified to the correct quality grade by performing the improved prediction method is expected to bring the effect of reducing production costs.

Influence Factor on Remediation of PAHs-Contaminated Soil by Using Flowing Subcritical Water (흐름식 아임계수를 이용한 PAHs 오염토양 정화 영향인자)

  • Jo, Young-Tae;Islam, Mohammad Nazrul;Park, Jeong-Hun
    • Journal of Soil and Groundwater Environment
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    • v.18 no.4
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    • pp.1-7
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    • 2013
  • Subcritical water which acts as organic solvent with increasing temperature and pressure because dielectric constant and viscosity decrease can be used to remediate PAHs-contaminated soil. Factors influencing on extraction were studied with varying the water temperature $200{\sim}275^{\circ}C$, extraction time 0~90 min, flow rate 10~100 mL/min and pressure 3.9~10MPa. 300 g of soil sample which was contaminated with PAHs(naphthalene, phenanthrene, fluoranthene and pyrene; 423, 420, 539 and 428 mg/kg of initial concentration) was packed into the cell and placed to reactor and then the subcritical water was pumped through the cell for PAHs extraction. Naphthalene was removed almost 100% at relatively low temperature ($200^{\circ}C$). The removal rate of phenanthrene, fluoranthene, and pyrene increased by 8, 26, and 23% when the temperature increased from 200 to $275^{\circ}C$; and it was gradually increased as extraction time increased from 0 to 90 min. Decreasing removal rate when water flow rate increased from 10 to 30 mL/min, but there was no significant change after 30 mL/min. This is supposed due to channeling phenomenon. The pressure was not an effective factor for extraction of PAHs in this study. Based on the results, the importance of effective factor was in following sequence: temperature >> time > flow rate.

Optimum Extraction Conditions and Anticancer Effect of Functional Polysaccharide from Mycelia of Grifola frondosa (잎새버섯(Grifola frondosa) 균사체의 기능성 다당류 최적 추출방법 및 항암효과)

  • Park, Chan-Ho;Lee, Gyeong-Min;Nam, Eun-Jeong;Yu, Yeon-Hee;Kim, Yong-Hyun;Kwon, Hyun-Jung;Yoon, Ok-Hyun;Han, Man-Deuk
    • The Korean Journal of Food And Nutrition
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    • v.25 no.1
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    • pp.181-187
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    • 2012
  • Grifola frondosa has been used as an herbal medicine for the treatment of cancer, diabetes mellitus and high blood pressure. In this study, functional polysaccharide was obtained from Grifola frondosa using four different extraction methods: hot water(HwFP), homogenize(HgFP), acid(AcFP), and alkali(AlFP) extraction methods. The effects of these extracts on KB and HepG2 cell lines were then examined for any anti-cancer activity. Alkaline extraction produced a yield of 0.175% and the total sugar content of the extract was 54.97%. We were able to confirm that the polysaccharide extracts from the mushroom produce an anti-cancer effect. The cytotoxicity of AlFP and AcFP against HepG2 cells were 22.86% and 28.88%, respectively, and the cytotoxicity of AlFP against the KB cell lines was 47.76% at a concentration of 1,000 ${\mu}g/m{\ell}$. Therefore, these results suggest that the optimum method for extracting functional polysaccharides from G. frondosa is the alkali extraction method.

Octimization of Conditions of Filtration and Concentration of Methanol Extract for Recovery of Paclitaxel from Plant Cell Culture (식물세포배양으로부터 Paclitaxel 회수를 위한 메탄올 추출액의 여과 및 농축 조건 최적화)

  • Kim, Jin-Hyun
    • KSBB Journal
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    • v.22 no.4
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    • pp.197-200
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    • 2007
  • This study examined the conditions of filtration and concentration of methanol extract from biomass. Filtration efficiency was improved by adding diatomaceous earth as a filter aid. The optimal amount of diatomaceous earth was 6% (w/w) to reduce the filtration time. The filtration time was reduced by 4.2% in first extraction, 30.0% in second extraction, 22.8% in third extraction, and 19.0% in fourth extraction, respectively. The optimal temperature of water bath was below 50$^{\circ}C$ for preventing paclitaxel degradation during concentration of methanol extract using a rotary evaporator. The temperature of concentrated solution in rotary evaporator was relatively low compared to bath temperature because of latent heat of evaporation. The stopping point of concentration in rotary evaporator for the following step was at a specific gravity of 0.96 of the concentrated solution in terms of the purity and yield of paclitaxel. This information is very useful for mass extraction of biomass for the recovery of paclitaxel from plant cell culture.

Protective effect of Salviae-radix extraction in $H_2O_2$ induced renal cell injury ($H_2O_2$에 의한 신장(腎臟) 세포 손상에 대한 단삼(丹參) 추출물의 방지 효과)

  • Kim, Sang-Beum;Jeong, Ji-Cheon
    • The Journal of Korean Medicine
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    • v.19 no.1
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    • pp.38-48
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    • 1998
  • This study was undertaken to determine whether Salviae-radix (SVR) extraction prevents the oxidant-induced cell injury and thereby exerts protective effect against oxidant-induced inhibition of tetraethylammonium uptake (TEA) in renal corticaJ sices. SVR (5%) attenuated $H_2O_2-induced$ inhibition of TEA uptake. $H_2O_2$ increased LDH release and lipid peroxidation in a dose-dependent manner. These changes were prevented by SVR extraction. The protective effect of SVR on LDH release was dose-dependent over the concentration range of 0.1-0.5%, and that on lipid peroxidation over the concentration ranges of 0.05-2%. SVR significantly prevented Hg-induced lipid peroxidation. SVR extraction (0.5%) increased cellular GSH content in normal and $H_2O_2-treated$ tissues. When slices were treated with 100 mM $H_2O_2$, catalase activity was decreased, which was prevented by 0.5% SVR extraction. The activity of glutathione peroxidase but not superoxide dismutase was significantly increased by 0.5% SVR extraction in $H_2O_2-treated$ tissuces. These results suggest that SVR has an antioxidant action and thereby exerts benefical effect against oxidant-induced impairment of membrane transport function. This effect of SVR is attributed to an increase in endogenous antioxidants such as GSH, catalase and glutathione peroxidase.

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