• Title/Summary/Keyword: Crystal violet

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Relationships between in-vitro virulence-associated characteristics, plasmid-bearing and production of Outer Membrane Protein(OMP) of Yersinia enterocolitica isolated from pigs (Yersinia enterocolitica의 시험관내 병원성 성상, plasmid 보유 및 외막 단백질(OMP) 생산간의 관계)

  • Park, Seog-gee;Choi, Chul-soon;Jeon, Yun-seong
    • Korean Journal of Veterinary Research
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    • v.32 no.2
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    • pp.181-194
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    • 1992
  • Two hundred and eighty nine strains of Yersinia enterocolitica isolated from healthy pigs were tested for the presence of 40~50 Megadalton virulence-associated plasmids and plasmidmediated in vitro virulence-associated properties, i.e., congo red uptake, calcium dependency, autoagglutination, CRMOX reaction, crystal violet binding and pyrazinamidase reaction. The correlationships between in vitro virulence-associated properties and the presence of 220 Kdalton outer membrane protein(OMP) were examined in strains with or without virulence-associated plasmids. The correlationships between the presence of plasmids on the production of the OMP and the expression of in vitro virulence-associated properties were studied with $CRMOX^+$ strains and acridine orangecured $CRMOX^-$ mutants. The results were as follows : 1. Of the in vitro virulence-associated tests with 289 strains of Y enterocolitica, 275 strains (95.2%) were positive for pyrazinamidase test, and followed by in order of crystal violet binding test, 226 (79.2% ) ; CRMOX test, 190 (65.7%) ; autoagglutination test, 1.85(64.0%) : calcium dependency test, 86 (29.8%) and congo red uptake test, 47(16.3%). 2. The correlationship between autoagglutination and CRMOX test(r=0.90) was highly significant (p<0.01). 3. In 190 strains(65.7%) bearing the virulence-associated plasmids(MW 40~50 Mdalton), the correlation between the presence of plasmids and their in vitro virulence-associated properties were highest with CRMOX test(r=0.93) and followed by in orders of AAG test(0.81), CV test(0.46), PYZ test(0.37) and CD test(0.18), but no correlationship between the presence of plasmids and CR test(-0.11). 4. The $CRMOX^+$ strains produced the 220 Kdalton OMP when they were cultured at $37^{\circ}C$, but not at $26^{\circ}C$. The presence of 220 Kdalton OMP was correlated significantly with in vitro virulence properties and the presence of virulence-associated plasmid, respectively. 5. In the isogenic $CRMOX^-$ mutant strains, of which plasmid were cured by treatment with acridine orange not only in vitro virulence-associated properties(CR 100%, CD 100%, AAG 82.6%, CV 58.3%) disappeared but also 220 Kdalton OMP(100%) was not produced. These results indicate that the positive CRMOX reaction is plasmid-mediated and the CRMOX test is potential as an in vitro virulence tests with Y enterocolitica.

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Inhibition of yeast Candida growth by protein antibiotic produced from Pseudomonas fluorescens BB2 (Pseudomonas fluorescens BB2 균주가 생산하는 단백질성 항생물질에 의한 효모 Candida 생육 억제)

  • Ahn, Kyung-Joon
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.448-452
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    • 2015
  • The bacterial strain that was isolated from chinese cabbage rhizosphere, showed inhibition of yeast growth. This strain was identified as Pseudomonas fluorescens BB2 by API 20NE test and 16S rRNA gene sequence analysis. P. fluorescens BB2 strain produced antibiotics against yeast as a secondary metabolite effectively when the culture was carried out in YM medium with 3% glucose at $20^{\circ}C$. The protein antibiotic of BB2 strain which was concentrated by ammonium sulfate precipitation and n-butanol extraction inhibited the growth of yeast with the minimal inhibitory concentration of $10{\mu}g/ml$ against Candida albicans KCTC 7965, and the growth of yeast was completely inhibited at $80{\mu}g/ml$. The hydrophilic fraction of n-butanol extraction inhibited the growth of Bacillus cereus ATCC 21366, showed orange halo on chrome azurol S plate, which means the fraction contained iron chelating siderophore. The results of crystal violet uptake through the cell membrane showed that membrane permeability was increased about 9% than control, when the concentration of hydrophobic antibiotic against yeast C. albicans was $60{\mu}g/ml$. As a result, the antibiotic produced by P. fluorescens BB2 against yeast Candida is considered antimicrobial peptide, and this is the first report in the genus Pseudomonas.

Molecular Characterization and Antimicrobial Susceptibility of Biofilm-forming Acinetobacter baumannii Clinical Isolates from Daejeon, Korea (대전지역에서 분리된 생물막 형성 Acinetobacter baumannii 임상분리주의 분자유전학적 특성과 항균제 감수성양상)

  • Sung, Ji Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.2
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    • pp.100-109
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    • 2018
  • The emergence and dissemination of multidrug-resistant (MDR) Acinetobacter baumannii isolates have been reported worldwide, with most of these possessing the ability to form biofilms. Biofilm formation is an important virulence factor associated with the resistance to disinfection and desiccation. This study examined the genetic basis of antimicrobial resistance mechanisms of biofilm-forming A. baumannii clinical isolates. Imaging and quantification of biofilms were performed by a crystal violet assay and 46 biofilm-forming A. baumannii isolates were selected. Subsequently, 16 isolates belonging to different clones were identified using REP-PCR, and detection of the antimicrobial determinants in the isolates was carried out. The 16 isolates included 9 non-MDR and 7 MDR isolates. The mean biomass $OD_{560}$ values of the non-MDR (0.96) and MDR (1.05) isolates differed but this difference was not significant. In this study, most biofilm-forming MDR A. baumannii isolates contained various antimicrobial resistance determinants ($bla_{OXA-23}$, armA, and mutations of gyrA and parC). On the other hand, most biofilm-forming non-MDR A. baumannii isolates did not contain antimicrobial resistance determinants. These results suggest that there is little correlation between the biofilm-forming ability and antimicrobial susceptibility in A. baumannii isolates. In addition, the emergence of MDR A. baumannii clinical isolates is generally caused by mutations of the genes associated with antimicrobial resistance and/or the acquisition of various antimicrobial resistance determinants.

Antifungal Activity of Rheum undulatum on Candida albicans by the Changes in Membrane Permeability (막투과성 변화로 인한 대황의 Candida albicans에 대한 항진균 활성)

  • Lee, Heung-Shick;Kim, Younhee
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.360-367
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    • 2014
  • Candida albicans is an opportunistic and the most prevalent fungal pathogen that can cause superficial and systemic infections in immunocompromised patients. C. albicans can promote the transition from budding yeast to filamentous form, generating biofilms. Infections associated with C. albicans biofilms are frequently resistant to conventional antifungal therapy. Therefore, the development of more effective antifungal drugs related with biofilm formation is required urgently. The roots of Rheum undulatum have been used for medicinal purposes in Korea and China traditionally. The aim of present study was to evaluate the effect of R. undulatum extract upon preformed biofilms of 12 clinical C. albicans isolates and the antifungal activities. Its effect on preformed biofilms was evaluated using XTT reduction assay, and metabolic activity of all tested strains was reduced significantly ($49.4{\pm}6.0%$) at 0.098 mg/ml R. undulatum. The R. undulatum extract blocked the adhesion of C. albicans biofilms to polystyrene surfaces, and damaged the cell membrane integrity of C. albicans which was analyzed by CFDA, AM, and propidium iodide double staining. It caused cell lysis which was observed by Confocal laser scanning and phase contrast microscope after propidium iodide and neutral red staining, respectively. Membrane permeability was changed as evidenced by crystal violet uptake. The data suggest that R. undulatum inhibits biofilm formation by C. albicans, which can be associated with the damage of the cell membrane integrity, the changes in the membrane permeability and the cell lysis of C. albicans.

Decolorization of Azo, Triphenylmethane and Heterocyclic Dyes by Irpex zonatus BN2 (송곳니구름버섯(Irpex zonatus) BN2에 의한 아조계, 트리페닐메탄계 및 헤테로싸이클릭계 염료의 탈색)

  • Yoon, Kyung-Ha;Choi, Yang-Soon
    • The Korean Journal of Mycology
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    • v.26 no.1 s.84
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    • pp.8-15
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    • 1998
  • The present research was undertaken to investigate the activity of ligninolytic enzymes and the decolorization capability of some dyes with Irpex zonatus BN2, isolated from nature and identified. For the assay of enzyme activities, the isolate did not produce lignin peroxidase (LiP) and veratryl alcohol oxidase (VAO), but laccase and manganese dependent peroxidase (MnP). While the activity for MnP was low $(61.6\;nmol/mg{\cdot}protein)$, its laccase activity was very high $(1185.9\;nmol/mg{\cdot}protein)$. Moreover, laccase had appeared earlier than MnP. When the isolate was incubated with each dye for 10 days, the decolorization rates of azo dyes, such as orange II, orange G, tropaeolin O and congo red were 98.0%, 97.4%, 99.0% and 95.3%, respectively. In case of heterocyclic dyes, eosin Y, toludine blue, methyl blue and azur B were 97.4 %, 98.7%, 99.9% and 94.0% respectively. Finally the results of triphenylmethane dye such as basic fuchsin, malachite green and crystal violet were 98.5%, 95.7% and 99.4%, respectively. The results suggest that laccase of Irpex zonatus BN2 should be played an important role in the decolorization of the dyes.

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Antibacterial effect of electrolyzed water on Streptococcus mutans (전기분해수소수의 Streptococcus mutans에 대한 항균효과)

  • Kim, Ji-Hye;Youn, Ha-Young;Kim, Eun-Kyong;Lee, Young-Eun;Jang, Ji-Eon;Song, Keun-Bae
    • Journal of Korean society of Dental Hygiene
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    • v.21 no.5
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    • pp.527-533
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    • 2021
  • Objectives: Electrolyzed water has been proven to have antibacterial effects against various microorganisms. However, there are only a few studies about effects of electrolyzed water on oral bacteria. The purpose of this study was to examine the antibacterial effect of electrolyzed water on Streptococcus mutans in vitro. Methods: S. mutans KCOM 1054 was treated with electrolyzed water for 1 or 3 minutes and plated on Mitis Salivarius agar with 15% sucrose and bacitracin. After incubation for 48 hours, colony forming units (CFU) were counted, and dental plaque was quantified by crystal violet staining. Results: The growth of S. mutans was significantly inhibited by electrolyzed water (p<0.001). In addition, the dental plaque formation by S. mutans was decreased in a time-dependent manner by exposure to electrolyzed water (p<0.001). Conclusions: Our results suggest that electrolyzed water can effectively prevent dental caries by inhibiting growth of (and the formation of dental plaque by) S. mutans.

Cytological Study on the Cause of the Osteoporotic Side Effects of Adefovir Dipivoxil (아데포비어의 부작용인 골다공증 원인 규명을 위한 세포학적 연구)

  • Park, Ho
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.379-385
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    • 2019
  • Osteoporosis is a disease that increases the risk of fractures by inducing a decrease in bone strength by the changes in hormones and a decrease in minerals. Recent reports have indicated that the long-term administration of Adefovir dipivoxil (ADV), which is used as a treatment for the hepatitis virus and AIDS, may have osteoporotic side effects. On the other hand, there are few studies on the cytopathic correlation of these causes. In this study, the biological relevance of ADV was evaluated using osteoblast hFOB1.19 and vascular endothelial cell HUVEC. First, the cells were treated with ADV at different concentrations, and DAPI and crystal violet staining were performed for morphological analysis of each cell and nucleus. A CCK-8 assay, real-time PCR, alkaline phosphatase (ALP) staining, and activity was performed to evaluate the drug effects on cell proliferation, gene expression, and osteoblast differentiation. As a result, ADV induced cell hypertrophy in hFOB1.19 cells and HUVEC cells. Furthermore, ADV not only inhibited cell proliferation and TGF-${\beta}$ expression but was also involved in osteoblast differentiation. Overall, these results provide basic data to help better understand the mechanism of ADV-induced osteoporosis and its clinical implications.

Effect of Adefovir Dipivoxil on the Inhibition of Osteogenic Differentiation of Mesenchymal Stem Cells and Osteoblasts (아데포비어가 중간엽 줄기세포와 조골세포의 골형성 분화 억제에 미치는 영향)

  • Ho PARK
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.4
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    • pp.284-290
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    • 2023
  • Adefovir dipivoxil (ADV) is used for the treatment of hepatitis and acquired immunodeficiency syndrome, but long-term use can cause osteoporosis. In this study, the effect of ADV on the osteocyte maturation process was evaluated at the level of undifferentiated cells using mesenchymal stem cells (MSCs) and osteoblasts (MG63). First, MSCs and MG63 cells were treated with ADV at different concentrations, and then a Cell Counting Kit-8 analysis was performed to determine the effect on the proliferation of each cell. Additionally, crystal violet and Hoechst staining were performed for the morphological analysis of each cell and nucleus. To determine the cause of cell hypertrophy, the transforming growth factor-beta (TGF-β) expression was investigated, and alkaline phosphatase (ALP) staining and activity were measured to determine the degree of differentiation of the MSCs and MG63 cells into mature osteocytes. The results confirmed that the ADV increases the expression of TGF-β in MSCs and MG63 cells, causing cellular and nuclear hypertrophy, and can cause osteoporosis by inhibiting cell proliferation and affecting the differentiation of mature osteocytes. Therefore, it is believed that these results can be used as a basis for understanding the adverse effects of ADV at a cytological level in basic medicine and clinical research.

A Study on the Luminescent Characteristics of YPO4:Pr3+ Phosphor by the Content Ratio of Pr6O11 and Calcination Temperature (Pr6O11의 함량 및 열처리 조건에 따른 YPO4:Pr3+ 형광체의 발광 특성 연구)

  • Min Jun Kim;Seong Eui Lee
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.37 no.1
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    • pp.68-73
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    • 2024
  • In this study, the praseodymium-doped yttrium phosphate (YPO4:Pr3+) powder, which is well known for its high luminescent efficiency, and long life in the UV range, was synthesized with various content ratios of Pr6O11 and calcination temperature. Crystal structure and luminescent properties of various phosphor powders based on different concentrations and calcination conditions were characterized by XRD (X-Ray Diffraction) and PL (photoluminescence) spectrometers. From the XRD analysis, the structure of YPO4:Pr3+ which is calcinated at 1,200℃ was stable tetragonal phase and crystal size was calculated about 25 nm by Scherrer equation. PL emission of YPO4:Pr3+ with a different content ratio of Pr6O11 by excitation λexc=250 nm shows that 0.75 mol% phosphor powder has maximum PL intensity and PL decreases with the increase of the ratio of Pr6O11 up to 1.25 mol% which is caused by changes of crystallinity of phosphor powders. With increasing dopant ratio, photo-luminescence Emission decreases due to Concentration quenching, which is commonly observed in phosphors. Currently, 0.75 mol% is considered the optimal doping concentration. A hybrid ultraviolet-emitting device incorporating YPO4:Pr3+ fluorescent material with plasma discharge was fabricated to enhance UV germicidal effects while minimizing ozone generation. UV emission from the plasma discharge device was shown at about 200 nm and 350 nm which caused additional emission of the regions of 250 nm, 315 nm, and 370 nm from the YPO4:Pr3+ phosphor.

Biological Manganese Removal in Water Treatment (정수처리에서 생물학적 망간처리)

  • Kim, Berm-Soo;Yoon, Jaekyung;Ann, Hyo-Won;Kim, Chung-Hwan
    • Journal of Korean Society of Water and Wastewater
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    • v.20 no.1
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    • pp.44-52
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    • 2006
  • Bio-filtration processes using honeycomb tubes (process 1) and aeration and manganese-sand filtration (process 2) were evaluated for the biological manganese removal efficiency. The concentration of manganese at effluent was stabilized after 20days operation in process 1. It was estimated the required time for attaching and growing microorganisms to honeycomb tubes. In long term of operation periods, manganese removal efficiency was dropped for the excessively attached biofilm and manganese dioxide to honeycomb tubes. It took several days for normal operation in process 2, after that manganese removal efficiency was increased to 98% and stabilized for 1.5 years. Microorganisms in process 1 and 2 were isolated and cultured to characterize manganese-oxidizing bacteria. Among the four types of colony, light brown colony was turned blue color by leuco crystal violet spot test. Stenotropomonas genus, known as manganese-oxidizing bacteria, was identified by 16S rDNA partial sequencing analysis which was isolated in process 1 and 2. For the biological treatment to remove manganese, these two considerations are important. One is to choose the proper media attaching manganese oxidant, another one is to define the cultural condition of isolated manganese-oxidizing bacteria.