• Title/Summary/Keyword: resistance detection

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Rapid detection of Rifampicin- resistant M, tuberculosis by PCR-SSCP of rpoB gene (결핵균의 rpoB유전자 PCR-SSCP법에 의한 Rifampicin 내성의 신속 진단)

  • Shim, Tae Sun;Yoo, Chul-Gyu;Han, Sung Koo;Shim, Young-Soo;Kim, Young Whan
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.842-851
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    • 1996
  • Background : Rifampicin(RFP) is a key component of the antituberculous shon-course chemotherapy and the RFP-resistance is a marker of multi-drug resistant(MDR) M. tuberculosis. rpoB gene encodes the ${\beta}$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. Recent reports show that rpoB gene mutations are the cause of RFP resistance of M. tuberculosis and the main mechanism of rpoB gene mutation is point mutation. And PCR-SSCP is a rapid and easy method for detecting point mutations. So we performed PCR-SSCP of rpoB gene of M. tuberculosis and compared the result with traditional RFP sensitivity test. Method : The 27 RFP sensitive M. tuberculosis culture isolates and 25 RFP resistant isolates were evaluated. The RFP sensitivity test was done at the Korean Tuberculosis istitute. The DNA was extracted by bead beater method and was amplified with primers TR-8 and TR-9 in a 20ul PCR reaction containing 0.1ul(luCi) [${\alpha}-^{32}P$] - dCTP. After amplification, SSCP was done using non-denaturaring polyacrylamide gel electrophoresis. Then direct sequencing was done in cases of different eletrophoretic mobility compared with that of H37Rv. In 19 cases, we compared PCR-SSCP results with patient's clinical course and the results of traditional RFP sensitivity test. Results : 1) All 27 RFP sensitive M. tuberculosis isolates showed the same electrophoretic mobility compared with that of H37Rv. And all 25 RFP resistant M. tuberculosis isolates showed different electrophoretic mobility. 2) The mechanism of rpoB gene mutation of M. tuberculosis is mainly point mutation. 3) The PCR-SSCP results correlate well with traditional RFP sensitivity and patient's clinical response to antituberculous treatment. Conclusion: The PCR-SSCP of rpoB gene is a very sensitive and rapid mehod in detecting RFP- resistant M. tuberculosis.

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Adhesion Characteristics and the High Pressure Resistance of Biofilm Bacteria in Seawater Reverse Osmosis Desalination Process (역삼투 해수담수화 공정 내 바이오필름 형성 미생물의 부착 및 고압내성 특성)

  • Jung, Ji-Yeon;Lee, Jin-Wook;Kim, Sung-Youn;Kim, In-S.
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.1
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    • pp.51-57
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    • 2009
  • Biofouling in seawater reverse osmosis (SWRO) desalination process causes many problems such as flux decline, biodegradation of membrane, increased cleaning time, and increased energy consumption and operational cost. Therefore biofouling is considered as the most critical problem in system operation. To control biofouling in early stage, detection of the most problematic bacteria causing biofouling is required. In this study, six model bacteria were chosen; Bacillus sp., Flavobacterium sp., Mycobacterium sp., Pseudomonas aeruginosa, Pseudomonas fluorescens, and Rhodobacter sp. based on report in the literature and phylogenetic analysis of seawater intake and fouled RO membrane. The adhesion to RO membrane, the high pressure resistance, and the hydrophobicity of the six model bacteria were examined to find out their fouling potential. Rhodobacter sp. and Mycobacterium sp. were found to attach very well to RO membrane surface compared to others used in this study. The test of hydrophobicity revealed that the bacteria which have high hydrophobicity or similar contact angle with RO membrane ($63^{\circ}$ of contact angle) easily attached to RO membrane surface. P. aeruginosa which is highly hydrophilic ($23.07^{\circ}$ of contact angle) showed the least adhesion characteristic among six model bacteria. After applying a pressure of 800 psi to the sample, Rhodobacter sp. was found to show the highest reduction rate; with 59-73% of the cells removed from the membrane under pressure. P. fluorescens on the other hand analyzed as the most pressure resistant bacteria among six model bacteria. The difference between reduction rates using direct counting and plate counting indicates that the viability of each model bacteria was affected significantly from the high pressure. Most cells subjected to high pressure were unable to form colonies even thought they maintained their structural integrity.

Virulence Profile and Antimicrobial Resistance of Escherichia coli from Flies Captured from Agricultural Environment (농업환경에 서식하는 파리에서 분리된 E. coli의 병원성 유전자 및 항생제 내성 조사)

  • Yun, Bohyun;Jang, Youn Jung;Kim, Yeon Rok;Kim, Hwang-Yong;Kim, Won-Il;Han, Sanghyun;Kim, Se-Ri;Ryu, Jae-Gee;Kim, Hyun Ju
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.147-153
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    • 2017
  • The purpose of this study was to isolate Escherichia coli from flies and to assess pathogenic genes and antibiotic resistance of the isolates. A total of 188 flies were captured in agricultural environment including fruits farms (n = 19), fermented soybean farms (n = 9), municipal waste (n = 46), livestock farms (n = 66), slaughterhouses (n = 38), and manure ground (n = 10). E. coli isolates of captured flies were tested for pathogenic gene and antibiotic resistance using PCR methods and VITEK2 systems. As a result, E. coli from 63% (119/188) of the captured flies has been detected, and the detection rate of E. coli was the highest (89%, 31/34) in flies captured at particular slaughterhouse. Of the 34 isolates, 94% (32/34) were pathogenic gene (ST gene) positive. Twenty-six percent (31/119) of the E. coli isolates were observed being resistant to one or more antibiotics. Markedly, one of E. coli isolates from Livestock farms was resistant to 7 antibiotics including ampicillin, ampicillin/sulbactam, cefazolin, cefotaxime, gentamicin, levofloxacin, and trimethoprim/sulfamethoxazole. In addition, it was ESBL positive. The results of the present study may suggest a risk of transmission of pathogenic and antimicrobial resistant bacteria from flies to livestock environment Therefore, it may need to prevent introducing flies into the agricultural production environment for safe food production.

Monitoring of Selected Veterinary Antibiotics in Animal Carcass Disposal Site and Adjacent Agricultural Soil (가축매몰지 및 인근 농경지의 축산용 잔류 항생제 모니터링)

  • Lim, Jung Eun;Rajapaksha, Anushka Upamali;Jeong, Se Hee;Kim, Sung Chul;Kim, Kye Hoon;Lee, Sang Soo;Ok, Yong Sik
    • Journal of Applied Biological Chemistry
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    • v.57 no.3
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    • pp.189-196
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    • 2014
  • In Korea, over 4,700 animal carcass disposal sites were installed until 2011 due to the outbreak of foot and mouth disease. Due to the putrefaction of buried animals, the leachate containing veterinary antibiotics may release into surrounding environments. Antibiotic residues in the environment cause the formation of antibiotic resistance bacteria threatening human and ecosystem health. This study reports the concentrations of five antibiotics, including tetracycline (TC), chlortetracycline (CTC), oxytetracycline (OTC), sulfamethazine (SMZ), and sulfamethoxazole (SMX), in soils from animal carcass disposal site and adjacent agricultural field. Concentrations of antibiotics at animal carcass disposal sites (TC: $144.26-350.73{\mu}g/kg$, SMZ: $17.72-44.94{\mu}g/kg$) were higher than those at agricultural field (TC: $134.16-320.73{\mu}g/kg$, SMZ: $6.48-8.85{\mu}g/kg$) whereas the concentrations of CTC, OTC, and SMX were below detection limit in both sites. Results showed that the antibiotics in animal carcass site might leach to the soil and possibly contaminating the groundwater. Future studies will focus on the transfer of antibiotics residues into food crops.

Public Health Risks: Chemical and Antibiotic Residues - Review -

  • Lee, M.H.;Lee, H.J.;Ryu, P.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.3
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    • pp.402-413
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    • 2001
  • Food safety is a term broadly applied to food quality that may adversely affect human health. These include zoonotic diseases and acute and chronic effects of ingesting natural and human-made xenobiotics. There are two major areas of concern over the presence of residues of antibiotics in animal-derived foodstuffs with regard to human health. The first is allergic reactions. Some antibiotics, such as penicillins can evoke allergic reactions even though small amounts of them are ingested or exposed by parenteral routes. The second is development of antibiotic resistance in gut bacteria of human. Recently multi-resistant pneumococcal, glycopeptide-resistant enterococci and gram negative bacteria with extended-spectrum $\beta$-lactamases have spread all over the world, and are now a serious therapeutic problem in human. Although it is evident that drugs are required in the efficient production of meat, milk and eggs, their indiscriminate use should never be substituted for hygienic management of farm. Drug should be used only when they are required. In addition to veterinary drugs, environmental contaminants that were contaminated in feed, water and air can make residues in animal products. Mycotoxins, heavy metals, pesticides, herbicides and other chemicals derived from industries can be harmful both to animal and human health. Most of organic contaminants, such as dioxin, PCBs and DDT, and metals are persistent in environment and biological organisms and can be accumulated in fat and hard tissues. Some of them are suspected to have endocrine disrupting, carcinogenic, teratogenic, immunodepressive and nervous effects. The governmental agencies concerned make efforts to prevent residue problems; approval of drugs including withdrawal times of each preparation of drugs, establishment of tolerances, guidelines regarding drug use and sanitation enforcement of livestock products. National residue program is conducted to audit the status of the chemical residues in foods. Recently HACCP has been introduced to promote food safety from farm to table by reducing hazardous biological, chemical and physical factors. Animal Production Food Safety Program, Quality Assurance Programs, Food Animal Residue Avoidance Databank are para- or non-governmental activities ensuring food safety. This topic will cover classification and usage or sources of chemical residues, their adverse effects, and chemical residue status of some countries. Issues are expanded to residue detection methodologies, toxicological and pharmacokinetic backgrounds of MRL and withdrawal time establishments, and the importance of non-governmental activities with regard to reducing chemical residues in food.

High Performance Flexible Inorganic Electronic Systems

  • Park, Gwi-Il;Lee, Geon-Jae
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.08a
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    • pp.115-116
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    • 2012
  • The demand for flexible electronic systems such as wearable computers, E-paper, and flexible displays has increased due to their advantages of excellent portability, conformal contact with curved surfaces, light weight, and human friendly interfaces over present rigid electronic systems. This seminar introduces three recent progresses that can extend the application of high performance flexible inorganic electronics. The first part of this seminar will introduce a RRAM with a one transistor-one memristor (1T-1M) arrays on flexible substrates. Flexible memory is an essential part of electronics for data processing, storage, and radio frequency (RF) communication and thus a key element to realize such flexible electronic systems. Although several emerging memory technologies, including resistive switching memory, have been proposed, the cell-to-cell interference issue has to be overcome for flexible and high performance nonvolatile memory applications. The cell-to-cell interference between neighbouring memory cells occurs due to leakage current paths through adjacent low resistance state cells and induces not only unnecessary power consumption but also a misreading problem, a fatal obstacle in memory operation. To fabricate a fully functional flexible memory and prevent these unwanted effects, we integrated high performance flexible single crystal silicon transistors with an amorphous titanium oxide (a-TiO2) based memristor to control the logic state of memory. The $8{\times}8$ NOR type 1T-1M RRAM demonstrated the first random access memory operation on flexible substrates by controlling each memory unit cell independently. The second part of the seminar will discuss the flexible GaN LED on LCP substrates for implantable biosensor. Inorganic III-V light emitting diodes (LEDs) have superior characteristics, such as long-term stability, high efficiency, and strong brightness compared to conventional incandescent lamps and OLED. However, due to the brittle property of bulk inorganic semiconductor materials, III-V LED limits its applications in the field of high performance flexible electronics. This seminar introduces the first flexible and implantable GaN LED on plastic substrates that is transferred from bulk GaN on Si substrates. The superb properties of the flexible GaN thin film in terms of its wide band gap and high efficiency enable the dramatic extension of not only consumer electronic applications but also the biosensing scale. The flexible white LEDs are demonstrated for the feasibility of using a white light source for future flexible BLU devices. Finally a water-resist and a biocompatible PTFE-coated flexible LED biosensor can detect PSA at a detection limit of 1 ng/mL. These results show that the nitride-based flexible LED can be used as the future flexible display technology and a type of implantable LED biosensor for a therapy tool. The final part of this seminar will introduce a highly efficient and printable BaTiO3 thin film nanogenerator on plastic substrates. Energy harvesting technologies converting external biomechanical energy sources (such as heart beat, blood flow, muscle stretching and animal movements) into electrical energy is recently a highly demanding issue in the materials science community. Herein, we describe procedure suitable for generating and printing a lead-free microstructured BaTiO3 thin film nanogenerator on plastic substrates to overcome limitations appeared in conventional flexible ferroelectric devices. Flexible BaTiO3 thin film nanogenerator was fabricated and the piezoelectric properties and mechanically stability of ferroelectric devices were characterized. From the results, we demonstrate the highly efficient and stable performance of BaTiO3 thin film nanogenerator.

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The Study of Near-field Scanning Microwave Microscope for the Nondestructive Detection System (비파괴 측정을 위한 근접장 마이크로파 현미경 연구)

  • Kim, Joo-Young;Kim, Song-Hui;Yoo, Hyun-Jun;Yang, Jong-Il;Yoo, Hyung-Keun;Yu, Kyong-Son;Kim, Seung-Wan;Lee, Kie-Jin
    • Journal of the Korean Society for Nondestructive Testing
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    • v.24 no.5
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    • pp.508-517
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    • 2004
  • We described a near-field scanning microwave microscope which uses a high-quality dielectric resonator with a tunable screw. The operating frequency is f=4.5 5GHz. The probe tip is mounted in a cylindrical resonant cavity coupled to a dielectric resonator We developed a hybrid tip combining a reduced length of the tapered part with a small apex. In order to understand the function of the probe, we fabricated three different tips using a conventional chemical etching technique and observed three different NSMM images for patterened Cr films on glass substrates. We measured the reflection coefficient of different metal thin film samples with the same thickness of 300m and compared with theoretical impedance respectly. By tuning the tunable screw coming through the top cover, we could improve sensitivity, signal-to-noise ratio, and spatial resolution to better than $1{\mu}m$. To demonstrate the ability of local microwave characterization, the surface resistance of metallic thin films has been mapped.

Presence of Carbonic Anhydrase III-like Protein in Shaggy Sea Raven, Hemitripterus villosus (삼세기(Shaggy sea raven, Hemitripterus villosus)의 carbonic anhydrase III에 관한 연구)

  • Kweon, Rok Eun;Kho, Kang Hee
    • Journal of Life Science
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    • v.24 no.2
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    • pp.186-190
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    • 2014
  • Carbonic anhydrase isozymes (CAs) are widespread zinc-containing metalloenzyme family. The enzyme catalyzes the reversible interconversion of $CO_2$ and $HCO_3$. This reaction is the main role of CA enzymes in physiological conditions. CA III, one of the CA isozymes, has been identified in many tissues. It is distinguished from the other isozymes by several characteristics, particularly by a lower specific activity and by its resistance to acetazolamide. However, the physiological function of CA III in fish is unknown. In this study, we examined the detection of CAs in the Shaggy sea raven Hemitripterus villosus, using SDS-PAGE, isoelectric focusing (IEF), and western blot analysis. We detected a significant protein band with molecular weight about 30 kDa from the tissues of H. villosus by SDS-PAGE and western blotting. A specific band of CA III with pI 7.0 was detected by IEF and western blotting in gill and muscle. The immunoreaction of anti-CA III expressed in the gill of H. villosus was much stronger than other tissues. One explanation for this result is that the fish gill is the only organ that is exposed to the external environment and that plays an important role in acid-base relevant ion transfer, the transfer of $H^+$ and/or $HCO{_3}^-$, for the maintenance of systemic pH. This is the first report on the identification of a carbonic anhydrase III-like protein from H. villosus.

Effect of Coexisting Ions on Electrokinetic Injection in Capillary Electrophoresis Analysis (모세관 전기영동 분석에서 계면 동전기 주입에 미치는 공존 이온의 영향)

  • Lee, Kwang-Woo;Jeon, Ji-Young;Lee, Kwang-Pill
    • Analytical Science and Technology
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    • v.9 no.1
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    • pp.35-42
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    • 1996
  • A rapid analytical method based on capillary electrophoresis is described for the determination of trace anions in high-purity chemicals which is used to prevent corrosion demage in nuclear power plants. Separations are carried out at 20kV using trimethylsilane-coated fused-silica capillary ($70cm{\times}50$ or $75{\mu}m$ i.d.) with the electrolyte of 5mM Chromate(pH=8). Detection was achieved using on-column indirect photometry at 254nm. The simultaneous analysis of inorganic anions, chloride, nitrate, sulfate, azide and phosphate was performed using methods of hydrodynamic(>1ppm) and / or electrokinetic(<1ppm) injection. The results of studies on the coexisting anions on analyte ions shows that peak responses of analyte in hydrodynamic injection is constant without effect of coexisting anions, but those of analysis in electrokinetic injection is strongly dependant upon the kind of coexisting anions and its ionic mobility. The analyte enrichment rate, hence peak response, is positive relationship with the resistance of the sample solution. Thus, appropriate measures, such as standard addition or internal standard technique, must be used to account for differences in conductance of standard and sample solutions.

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Prevalence and Molecular Characterization of Methicillin Resistant Staphylococcus aureus Isolated from Raw Milk Samples in Gyeonggi-do, Korea (원유시료 중 메티실린 내성 황색포도알균의 분포 및 내성 유전자 특성 분석)

  • Kang, SoWon;Song, YoungCheon;Choi, SungSook
    • Journal of Food Hygiene and Safety
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    • v.29 no.3
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    • pp.223-227
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    • 2014
  • This study was investigated to determine the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) isolated from raw milk samples and to further study on the molecular characteristics of the MRSA isolates. Using Staphylococcus Medium 110, Staphylococcus spp. were isolated from raw milk samples and further identification was carried by Vitek2 system. Minimum inhibitory concentrations (MICs) of antibiotics were conducted by serial dilution method according to the Clinical Laboratory Standards Institute (CLSI) guideline. For the detection of resistance genes and molecular characterization, PCR reaction was performed by gene specific primers and followed by DNA sequencing. Of the 698 milk samples, 94 Staphylococcus aureus (S. aureus) were identified (94 S. aureus/286 Staphylococcus spp.). Of the 94 S. aureus, seven isolates have mecA, a methicillin resistant gene. mecA positive seven isolates were then characterized by staphylococcal cassette chromosome mec (SCCmec) typing, and Panton-Valentine Leukocidin (pvl) gene using PCR. All of mecA positive isolates were resistant to ampicillin and oxacillin, but sensitive to teicoplanin, vancomycin and ciprofloxacin. One of seven isolates was SCCmec type II and six isolates were type IV and all seven isolates were pvl gene negative.