• Title/Summary/Keyword: Cell density

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An ELISA-on-a-Chip Biosensor System for Early Screening of Listeria monocytogenes in Contaminated Food Products

  • Seo, Sung-Min;Cho, Il-Hoon;Kim, Joo-Ho;Jeon, Jin-Woo;Oh, Eun-Gyoung;Yu, Hong-Sik;Shin, Soon-Bum;Lee, Hee-Jung;Paek, Se-Hwan
    • Bulletin of the Korean Chemical Society
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    • v.30 no.12
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    • pp.2993-2998
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    • 2009
  • An enzyme-linked immunosorbent assay (ELISA)-on-a-chip (EOC) biosensor combined with cell concentration technology based on immuno-magnetic separation (IMS) was investigated for use as a potential tool for early screening of Listeria monocytogenes (L. monocytogenes) in food products. The target analyte is a well-known pathogenic foodborne microorganism and outbreaks of the food poisoning typically occur due to contamination of normal food products. Thus, the aim of this study was to develop a rapid and reliable sensor that could be utilized on a daily basis to test food products for the presence of this pathogenic microorganism. The sensor was optimized to provide a high detection capability (e.g., 5.9 ${\times}\;10^3$ cells/mL) and, to eventually minimize cultivation time. The cell density was condensed using IMS prior to analysis. Since the concentration rate of IMS was greater than 100-fold, this combination resulted in a detection limit of 54 cells/mL. The EOC-IMS coupled analytical system was then applied to a real sample test of fish intestines. The system was able to detect L. monocytogenes at a concentration of 2.4 CFU/g after pre-enrichment for 6 h from the onset of cell cultivation. This may allow us to monitor the target analyte at a concentration less than 1 CFU/g within a 9 h-cultivation provided a doubling time of 40 min is typically maintained. Based on this estimation, the EOC-IMS system can screen and detect the presence of this microorganism in food products almost within working hours.

A Study on the Life Time Prediction and Acid-Heat aging Property of NBR Rubber for Fuel Cell Gasket (연료전지 카스켓용 NBR 고무의 산-열 노화 특성과 수명예측에 관한 연구)

  • Kim, Mi-Suk;Kim, Jin-Hak;Kim, Seok-Jin;Kim, Jin-Kuk
    • Elastomers and Composites
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    • v.42 no.1
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    • pp.20-31
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    • 2007
  • Material characteristics and lifetime evaluation are very important in design procedure to assure the safety and reliability of the rubber components. In this paper, the NBR compound was prepared by sulfur-cure system, and was used in predicting the lifetime of rubber gasket made by the compound. The accelerated material aging was investigated at different temperatures at 120, 140 and $160^{\circ}C$ and aging time from 3 hours to 600 hours at 5, 6, 7 vol %. of $H_2SO_4$ concentrations The rubber strips were placed in acid solution using pyrex g1ass tube. Both ends of pyrex g1ass tube were sealed to avoid evaporation of solution during heating at given time. The material test and accelerated acid-heat aging test were carried out to predict the useful life of NBR rubber gasket for a fuel cell stack. In order to investigate the effects of acid-heat aging on the properties of the NBR, tensile strength, elongation at break, hardness and crosslink-density were measured. The tensile strength decreases as the $H_2SO_4$ concentrations and temperature increase. Results were evaluated using Arrhenius equation.

Characteristics of Growth and Metal Removal in Recombinant Saccharomyces cerevisiae harboring a Metallothionein Gene (Metallothionein 유전자가 도입된 재조합 Saccharomyces cerevisiae의 생육과 금속제거에 대한 특성)

  • 정동환;김대옥서진호
    • KSBB Journal
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    • v.10 no.5
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    • pp.475-481
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    • 1995
  • The effect of metallothionein expression on the metal resistance and removal by recombinant Saccharomyces cerevisiae containing the plasmid pJW9 was investigated. The recombinant strain S. cerevisiae BZ-pJ was constructed by transforming the host strain S. cerevisiae BZ3l-1-7Ba with the gene coding for a metal-binding protein, metallothionein. Introduction of the MT gene yielded an increase in the minimum inhibitory concentration (MIC) of copper more than three times compared with the host strain. The minimum inhibitory concentrations of $Cr^{2+}, Znr^{2+} and Pb^{2+}, $ were not different for the two strains. The recombinant yeast grown in a medium containing 8mM CuSO4 was able to remove copper with a capacity of 18.9mg $Cu^{2+}$/g dry cell. In a mixture of copper and zinc, the presence of copper relieved the toxic effects caused by zinc, resulting in an enhancement of the final cell density and the specific growth rate of the recombinant yeast. The capability to remove copper by the recombinant yeast was linearly proportional to the copper concentrations in the medium. The efficiency of copper removal was rather constant regardless of the initial copper concentrations. The specific removal of zinc was dependent on the zinc concentrations in media, though, and such dependence was not so pronounced as the concentration of copper.

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AN ELECTRONMICROSCOPIC STUDY ON THE HEALING PROCESS OF THE REMAINING PULPAL TISSUES AFTER PULPOTOMY BY Nd-YAG LASER (Nd-YAG레이저에 의한 치수 절단후 잔존 치수 조직의 치유과정에 관한 전자현미경적 연구)

  • Park, Dong-Sung;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.20 no.2
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    • pp.399-422
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    • 1995
  • The purpose of this study was to investigate a fragment of possibility of pulpotomy with the Nd-YAG laser by the observation of pulpal healing process and the fine structural changes of the fibroblasts of the remaining pulpal tissues. Class V cavities on !55 teeth from 4 adult dogs were prepared and the pulp chambers were opened with a sterilized round bur. In the control group(19 teeth), the exposed coronal pulps were excised by a sharp excarvator. After bleeding was controlled with the sterilized cotton pellets, calcium hydroxide powder was applied on the remaining pulpal tissues and the cavities were sealed with Z.O.E. In the experimental group 1 : the pulpotomy with laser-calcium hydroxide powder application group(l9 teeth), the exposed coronal pulps were excised by Nd-YAG laser(10 watts power, 2 psi water, 20 psi air) for 2 or 3 seconds and calcium hydroxide powder was applied on the remaining pulpal tissues and the cavities were sealed with Z.O.E. In the experimental group 2 : the pulpotomy with laser-no calcium hydroxide powder application group(17 teeth), after amputating the coronal pulps with Nd-YAG laser as the experimental group 1, the remaining pulpal tissues were covered with stenilized aluminum foil and the cavities were filled with Z.O.E. The animals were sacrificed at the intervals of 1, 2, 3 and 4 weeks. All the teeth were rouutinely processed and the remaining pulpal tissues were observed by the light microscope and electron microscope. The results were as follows : 1. In light microscopic findings, there was no significant difference of the inflammatory response in the remaining pulpal tissues between the control group and the experimental groups. In both of the experimental group 1 : pulpotomy with laser-calcium hydroxide powder application group and the control group, the dentin bridges were observed after 2 weeks and the structure of the dentin bridge was almost same. In the experimental group 2 : pulpotomy with laser-no calcium hydroxide powder application group, the fibrous layers instead of dentin bridge were observed on the superficial portion of the remaining pulpal tissues after 2 weeks and they were consisted with densely crowded active fibroblasts. 2. In the electronmicroscopic findings, the active fibroblasts in the experimental groups were more frequently observed than in the control group at 1 week. But active fibroblasts were found with same frequency after 2 weeks in all of the control group and the experimental groups. 3. General distortions of the cell such as loss of the cell membrane, vaculoization of the cell etc. were observed at the suberficial layer of the remaining pulpal tissues and the carbonization was found in the dentinal wall in 1 week of the experimental groups. 4. In the experimental group 2 : pulpotomy with laser-no calcium hydroxide powder application group, the activity and the density of the fibroblasts in the fibrous layer were more than those in the deep portion of the remaining pulpal tissues after 2 weeks. 5. In the control group, bacteria such as cocci and bacilli were observed frequently, but in the experimntal groups, they could not be observed.

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Biocompatibility of experimental mixture of mineral trioxide aggregate and glass ionomer cement (Glass ionomer cement와 혼합한 mineral trioxide aggregate의 생체친화성)

  • Oh, Min-Jae;Jeong, Yu-Na;Bae, In-Ho;Yang, So-Young;Park, Bum-Jun;Koh, Jeong-Tae;Hwang, Yun-Chan;Hwang, In-Nam;Oh, Won-Mann
    • Restorative Dentistry and Endodontics
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    • v.35 no.5
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    • pp.359-367
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    • 2010
  • Objectives: The purpose of the present in vitro study was to evaluate the biocompatibility of mineral trioxide aggregate (MTA) mixed with glass ionomer cement (GIC), and to compare it with that of MTA, GIC, IRM and SuperEBA. Materials and Methods: Experimental groups were divided into 3 groups such as 1 : 1, 2 : 1, and 1 : 2 groups depending on the mixing ratios of MTA powder and GIC powder. Instead of distilled water, GIC liquid was mixed with the powder. This study was carried out using MG-63 cells derived from human osteosarcoma. They were incubated for 1 day on the surfaces of disc samples and examined by scanning electron microscopy. To evaluate the cytotoxicity of test materials quantitatively, XTT assay was used. The cells were exposed to the extracts and incubated. Cell viability was recorded by measuring the optical density of each test well in reference to controls. Results: The SEM revealed that elongated, dense, and almost confluent cells were observed in the cultures of MTA mixed with GIC, MTA and GIC. On the contrary, cells on the surface of IRM or SuperEBA were round in shape. In XTT assay, cell viability of MTA mixed with GIC group was similar to that of MTA or GIC at all time points. IRM and SuperEBA showed significantly lower cell viability than other groups at all time points (p < 0.05). Conclusions: In this research MTA mixed with GIC showed similar cellular responses as MTA and GIC. It suggests that MTA mixed with GIC has good biocompatibility like MTA and GIC.

Application of Species-specific DNA Probe to Field Samples of Alexandrium tamarense (Lebour) Balech (자연 시료로부터 Alexandrium tamarense을 위한 종 특이적 DNA탐침의 응용)

  • Cho, Eun-Seob;Kim, Gi-Young;Park, Hyung-Sik;Kim, Hak-Gyoon;Moon, Sung-Ki;Lee, Jae-Dong
    • Journal of Life Science
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    • v.12 no.3
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    • pp.250-255
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    • 2002
  • Fluorescent species-specific DNA probe (AT1) of toxic dinoflagellate Arexandrium tamarense was tested on several other species, on comparison of binding activity at different preservatives for fixation of the cells, at different culture age and estimation of cell density by light microscope or epifluorescent microscope using whole cell hybridization. Th AT1 probe specifically bound to Alexandrium tamarense, whereas it did not bind to other phytoplankton, in particular Alexandrium catenella, morphologically similar to Alexandrium tamarense, could not react to AT1 probe. When cells were fixed with all three preservatives, labeling cells of Alexandrium tamarense emitted strong fluorescent signal intensity. In addition, regardless culture days, binding activity with AT1 probe was strong. The tell densities estimated by epifluorescent microscope were than those estimated by light microscope. The enumeration and identifying of Arexandriurn tamarense using DNA probe method will be contributed to a new biotoxin monitoring and prediction system in field.

Lactobacillus plantarum APsulloc 331261 Fermented Products as Potential Skin Microbial Modulation Cosmetic Ingredients (Lactobacillus plantarum APsulloc 331261 발효 용해물의 피부 미생물 조절 효과)

  • Kim, Hanbyul;Myoung, Kilsun;Lee, Hyun Gee;Choi, Eun-Jeong;Park, Taehun;An, Susun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.1
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    • pp.23-29
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    • 2020
  • The skin is colonized by a large number of microorganisms with a stable composition of species. However, disease states of skin such as acne vulgaris, psoriasis, and atopic dermatitis have specific microbiome compositions that are different from those of healthy skin. The target modulation of the skin microbiome can be a potential treatment for these skin diseases. Quorum sensing (QS), a bacterial cell-cell communication system, can control the survival of bacteria and increase cell density. Also, QS affects the pathogenicity of bacteria such as biofilm formation and protease production. In this study, we confirmed anti-QS activity of Amorepacific patented ingredients, which are Lactobacillus ferment lysate (using Lactobacillus plantarum APsulloc 331261, KCCM 11179P) through bio-reporter bacterial strain Chromobacterium violaceum. The purple pigment production of C. violaceum controlled by QS was reduced 27.3% by adding 10 ㎍/mL of Lactobacillus ferment lysate (freeze dried). In addition, the Lactobacillus ferment lysate increased growth of Staphylococcus epidermidis 12% and decreased growth of Pseudomonas aeruginosa 38.5% and its biofilm formation 17.7% at a concentration of 10 ㎍/mL compared to the untreated control group. Moreover, S. epidermidis was co-cultured with the representative dermatological bacterium Staphylococcus aureus in the same genus, the growth of S. epidermidis was increased 134 % and the growth of S. aureus was decreased 13%. These results suggest that fermented lysate using Lactobacillus plantarum APsulloc 331261 may be useful as a cosmetic ingredient that can control the balance of skin microbiome.

Toxic dinoflagellate Gymnodinium catenatum Graham(Dinophyceae) from the southern coast of Korea: morphology, phylogeny and effects of temperature and salinity on growth (남해안에서 분리한 유독 와편모조류 Gymnodinium catenatum Graham (Dinophyceae): 형태, 분자계통학적 특성 및 온도와 염분에 따른 성장 특성)

  • Han, Kyong Ha;Li, Zhun;Kang, Byeong Jun;Youn, Joo Yeon;Shin, Hyeon Ho
    • Korean Journal of Environmental Biology
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    • v.37 no.1
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    • pp.31-41
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    • 2019
  • The toxic dinoflagellate Gymnodinium catenatum isolated from the southern coast of Korea was described under light and scanning electron microscopy, and its large subunit (LSU) rDNA was sequenced. In addition, the effects of temperature and salinity on its growth were investigated. The cells of G. catenatum, as viewed under the electronic microscope, were green-brown color, $38.1-77.4{\mu}m$ in length and $26.1-40.8{\mu}m$ in width. The epicone was conical, while the hypocone was trapezoidal. The nucleus was located at the central part of the cell. The apical groove was horseshoe-shaped and small pores were irregularly distributed on the cell surface. Molecular phylogeny based on LSU rDNA gene sequences showed that the Korean G. catenatum and previously reported species formed a monophyletic clade within Gymnodinium sensu stricto clade. The maximum growth rate of $0.37day^{-1}$, was obtained at $25^{\circ}C$ and 35 psu, and the maximum cell density of $1,073cells\;mL^{-1}$, was observed at $20^{\circ}C$ and 25 psu. However, G. catenatum did not grow at temperature < $15^{\circ}C$ and < $30^{\circ}C$. These results suggest that environmental conditions of summer and autumn in the southern coast of Korea may be favorable for the growth of G. catenatum.

Growth of Candida albicans Biofilm is Inhibited by Salvia miltiorrhiza (단삼에 의한 Candida albicans 바이오필름 발달의 억제)

  • Lee, Heung-Shick;Kim, Younhee
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.465-472
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    • 2019
  • Candida albicans is an opportunistic human pathogen that causes infections. Candidiasis is often related to antifungal resistance because the pathogen has the ability to form biofilms. In a previous study, we found that the Salvia miltiorriza ethanol extract demonstrated anticandidal activity by altering membrane permeability and inhibiting the cell wall synthesis in C. albicans. Our results here demonstrate that $78{\mu}g/ml$ of the S. miltiorriza extract significantly diminished the early stage biofilms formed by 10 clinical C. albicans isolates by 51.3%; this was analyzed by 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt (XTT) reduction assay. The effect of the S. miltiorrhiza extract on the adhesion of C. albicans cells to polystyrene plates and germ tube formation was examined via microscopic investigation. Although the density of the adhered cells was remarkably reduced up on incubation with $39{\mu}g/ml$ S. miltiorrhiza extract, germ tube formation by C. albicans was rarely affected. Quantitative real-time PCR analysis showed that the S. miltiorrhiza extract downregulated the expression of C. albicans hypha-specific genes, EAP1 by 34.7% (p < 0.001), ALS1 by 45.0% (p < 0.001), ALS3 by 48.1% (p < 0.001), and ECE1 by 21.3% (p = 0.006), respectively. Our data suggest that the S. miltiorrhiza ethanol extract significantly inhibited the early stage of biofilm formation by C. albicans by interfering with cell adhesion, by downregulating EAP1, ALS1 and ALS3, and presumably by modifying the cell wall and membrane structure.

Enhancement of Electrochemical Performance of Cathode by Optimizing Laccase-Carbon Nanotubes Layers for Enzymatic Fuel Cells (Laccase-탄소나노튜브 적층을 통한 효소 연료전지의 cathode 성능 향상)

  • Wang, Xue;Kim, Chang-Joon
    • Korean Chemical Engineering Research
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    • v.60 no.4
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    • pp.550-556
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    • 2022
  • The performance of enzymatic fuel cells that convert chemical energy contained in various organic molecules such as sugar, alcohol, organic acids, and amino acids into electrical energy is greatly affected by the cathode as well as the anode. This study aimed to develop a laccase-based cathode with high performance. An enzyme composite composed of an laccase, redox mediator, and carbon nanotubes was immobilized on the surface of electrode in multiple layers, and the effect of the number of layers and the presence or absence of carbon nanotubes on electrode performance was investigated. As the number of layers of the enzyme-mediator (Lac-(PVI-Os-dCl)) on the electrode surface increased, the amount of reduction current generated at the electrode increased. The enzyme-carbon nanotube-mediator composite electrode (Lac-SWCNTs-(PVI-Os-dCl)) generated a current 1.7 times greater than that of the Lac-(PVI-Os-dCl). It was found that the largest amount of current (10.1±0.1 µA) was generated in the electrode composed of two layers of Lac-(PVI-Os-dCl) and two layers of Lac-SWCNTs-(PVI-Os-dCl) in the evaluation of electrodes with different ratio of Lac-SWCNTs-(PVI-Os-dCl) and Lac-(PVI-Os-dCl). The maximum power density of the cell using the cathode composed of a single layer of Lac-(PVI-Os-dCl) and the cell using the optimized cathode were 0.46±0.05 and 1.23±0.04 µW/cm2, respectively. In this study, it was demonstrated that the performance of cathode and the enzymatic fuel cell using the same can be improved by optimizing the layers of composites composed of laccase, redox mediator, and carbon nanotubes on the electrode surface.