• Title/Summary/Keyword: E.coil

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In silico annotation of a hypothetical protein from Listeria monocytogenes EGD-e unfolds a toxin protein of the type II secretion system

  • Maisha Tasneem;Shipan Das Gupta;Monira Binte Momin;Kazi Modasser Hossain;Tasnim Binta Osman;Fazley Rabbi
    • Genomics & Informatics
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    • v.21 no.1
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    • pp.7.1-7.11
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    • 2023
  • The gram-positive bacterium Listeria monocytogenes is an important foodborne intracellular pathogen that is widespread in the environment. The functions of hypothetical proteins (HP) from various pathogenic bacteria have been successfully annotated using a variety of bioinformatics strategies. In this study, a HP Imo0888 (NP_464414.1) from the Listeria monocytogenes EGD-e strain was annotated using several bioinformatics tools. Various techniques, including CELLO, PSORTb, and SOSUIGramN, identified the candidate protein as cytoplasmic. Domain and motif analysis revealed that the target protein is a PemK/MazF-like toxin protein of the type II toxin-antitoxin system (TAS) which was consistent with BLASTp analysis. Through secondary structure analysis, we found the random coil to be the most frequent. The Alpha Fold 2 Protein Structure Prediction Database was used to determine the three-dimensional (3D) structure of the HP using the template structure of a type II TAS PemK/MazF family toxin protein (DB ID_AFDB: A0A4B9HQB9) with 99.1% sequence identity. Various quality evaluation tools, such as PROCHECK, ERRAT, Verify 3D, and QMEAN were used to validate the 3D structure. Following the YASARA energy minimization method, the target protein's 3D structure became more stable. The active site of the developed 3D structure was determined by the CASTp server. Most pathogens that harbor TAS create a crucial risk to human health. Our aim to annotate the HP Imo088 found in Listeria could offer a chance to understand bacterial pathogenicity and identify a number of potential targets for drug development.

Inactivation of Pathogenic Bacteria by Addition of Thermophilic Bacteria in the Thermophilic Aerobic Oxidation(TAO) System (고온호기산화장치의 고온미생물 첨가에 의한 병원성 미생물의 불활성화)

  • Lee W. I.;H. Tsujii;T. Maki;Lee M. G.
    • Journal of Animal Environmental Science
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    • v.10 no.2
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    • pp.111-118
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    • 2004
  • This study analyzed temperature increase, microorganism changes, and inactivation of pathogenic microorganisms in pig slurry when treated with thermophilic microorganisms in Thermophilic Aerobic Oxidation(TAO) system. An amount of $6 m^3$ of pig slurry was treated in an $18 m^3(3.0\times2.5\times2.4 m)$ reactor for 5 to 7 days in two groups: the control of pig slurry only and the treatment of pig slurry with 6 liters of thermophilic microorganism(Bacillus sp.). To study the microorganism changes in the reactor, the populations of aerobic mesophilic microorganisms, thermophilic microorganisms and general pathogens were analyzed. To study the inactivation of pathogenic microorganisms, the levels of E. coli, Salmonella sp, Crytosporidium parvum and Giardia lamblia were analyzed. The temperature inside the reactor ranged from 18 to $62^{\circ}C$ for the control while far the treatment group it ranged from 18 to $66^{\circ}C$, showing a slightly higher array. With regard to changes in microorganisms, both mesophilic and thermophilic organisms decreased from $3.1\times10^6$ to $1.2\times10^2$ CFU/ml and from $1.0\times10^4$ to $8.0\times10^1$ CFU/ml, respectively, in the control. In the treatment, on the other hand, mesophilic organisms decreased from $3.0\times10^8$ CFU/ml to $8.6\times10^5$ CFU/ml while thermophilic organisms increased sharply from $2.0\times10^6$ to $1.2\times10^8$ CFU/ml. For pathogens, Salmonella and Giardia were not detected either before or after the treatment, while E. coli and C. parvum were found to be $10^5$ CFU/ml each before treatment and negative after it. From this experiment, it was concluded that thermophilic microorganisms could effectively sanitize liquid compost by generating high temperature in the TAO system, which in turn would inhibit the growth of pathogenic organisms.

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Method of Nitrate Nitrogen Determination for Plant, Soil and Water Analysis by E. coil Cells (E. coli 세포(細胞)를 이용한 식물(植物), 토양(土壤) 및 수질(水質)의 질산태(窒酸態) 질소(窒素) 분석방법(分析方法))

  • Sohn, Sang Mok;Kucke, Martin;Lee, Yoon Gun
    • Korean Journal of Soil Science and Fertilizer
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    • v.30 no.4
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    • pp.361-369
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    • 1997
  • A microbiological nitrate determination method by E. coli is modified in Korea, using K12 wildtype, KCTC 1116, for the quantitative reduction of $NO_3{^-}$ to $NO_2{^-}$. The nitrate in plant, soil or water sample is determined spectrophotometrically after being diazotized with sulfaniamide and N-(1-naphthl)-ethlenediamine. The modified E. coli cell method and principle for nitrate determination using Korean wildtype E. coli strain is described, and cell culture and preparation of stock suspension for E. coli as well. This modified E. coli cell method can be managed simply and fast, it is suitable for the investigation of the large serials, it can be also automated and has a high degree of sensitivity up to 0.01ppm $NO_3{^-}-N$ in the sample solution. The applicability of the modified E. coli cell method has been tested for plant, soil and water analysis on a wide range of different samples. Recovery rates of added nitrate have been determined and comparisons with other standard nitrate analytical procedures have been carried out. The results with the modified E. coli cell method show high correlation ($r^2=0.98$) with those gained by the standard analytical procedures. The advantages and disadvantages of the method are also discussed to other nitrate determination methods.

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Incidence of Escherichia coli and Its Susceptibility to Antimicrobials in Childhood Urinary Tract Infection (소아 요로감염에서 Escherichia coli의 빈도와 항생제 감수성에 대한 연구)

  • Chi Hye-Mi;Kwahk Jae-Hyok;Lee Jun-Ho;Park Hye-Won
    • Childhood Kidney Diseases
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    • v.10 no.1
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    • pp.18-26
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    • 2006
  • Purpose : Empirical antimicrobial treatment is indicated before bacteriological results are available for young children with febrile UTI to minimize renal scarring. To ensure appropriate therapy, knowledge of the prevalence of causative organisms and their susceptibility patterns to antimicrobials is mandatory. We performed a retrospective analysis investigating the local prevalence and resistance patterns of uropathogens, primarily E. coli, isolated from community-acquired UTIs. Methods : A total of 103 positive urine cultures from children with febrile UTI collected at Bundang CHA General Hospital from February 2004 to February 2005 were analyzed. Inclusion criteria were fever higher than $37.5^{\circ}C$, significant bacteriuria with single strain growth of at least 10s colony forming units/mL urine, and leukocyturia >5/HPF. Results : E. coli(89.3%) was the leading uropathogen followed by Enterococcus spp.(3.9%) Klebsiella spp.(2.9%), Citrobctcter spp.(1.9%) and Enterobacter spp.(1.9%). E. coli strains revealed a low proportion of antimicrobial susceptibility to ampicillin(AMP; 27.2%) ampicillinsulbactam(AMS; 34.8%) and trimethoprim-sulfamethoxazole(SXT; 65.2%). Susceptibility patterns to cephalosporins were as follows; cefazolin(1st generation; 91.3%), cefoxitin(2nd; 100%), ceftriaxone(3rd; 97.8%) and cefepime(4th; 97.8%). Three E. coli isolates produced ex tended - spectrum beta-lactamase(ESBL). Conclusion : Empirical treatment with AMP, AMS and SXT, which are commonly used in pediatric clinics, is not recommended for childhood UTI due to high incidence of resistance. The high level of susceptibility to cephalosporins makes these drugs reasonable alternatives. However the emergence of ESBL-producers, even though they are quite few, may have an impact on cephalosporin treatment in the future. (J Korean Soc Pediatr Nephrol 2006;10:18-26)

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Microbial Prevalence and Quality of Organic Farm Produce from Various Production Sites (생산지 수집 신선 유기농 농산물 미생물 분포도 분석)

  • Park, Won-Jung;Ryu, Hwa-Yeon;Lim, Ga-Yeon;Lee, Young-Duck;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.46 no.2
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    • pp.262-267
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    • 2014
  • To analyze the presence of microbes in organic farm produce, green chillies, lettuce, tomatoes, apples, pears, and rice were collected at 47 production sites with organic and conventional produce. Total average bacterial counts of 4.07 log CFU/g in organic green chillies, 3.71 log CFU/g in conventional green chillies, and 6.76- 6.90 log CFU/g in the both lettuce were detected. Mean bacterial counts of 4.48 log CFU/g and 2.84 log CFU/g were detected in organic and conventional pear produce, respectively. Differences in bacterial counts in tomatoes, apples, and rice in organic and conventional produce were less pronounced. Clostridium perfringens, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., and Staphylococcus aureus were not detected in any produce sample. However, Bacillus cereus was detected with average counts of 1.04 log CFU/g in 11/47 (23%) conventional produce samples and 1.97 log CFU/g in 6/47 (13%) organic produce samples. Therefore, organic and conventional produce showed similar microbial prevalence patterns, and comparable safety in terms of pathogen contamination.

Fabrication of RF Inductor Using FeTaN Patterned Soft Magnetic Films (Patterned FeTaN 연자성 박막을 이용한 RF inductor의 제조)

  • Bae, Seok;Kim, Choong-Sik;Ryu, Sung-Ryong;Nam, Seoung-Eui;Kim, Hyoung-June;Song, Jae-Sung;Yamaguchi, Masahiro
    • Journal of the Korean Magnetics Society
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    • v.11 no.6
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    • pp.239-244
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    • 2001
  • Recently, RF inductor having researched by many workers, we fabricated and investigated properties of RF inductors. In order to improve the Q-factor (Quality), we try to apply the patterned Fe$_{78.81}$Ta$_{8.47}$N$_{12.71}$ soft magnetic thin film of 5000 which shows magnetic anisotropy of 30 Oe. Thus, patterned magnetic film was artificially increased magnetic anisotropy lead to increasing of ferro-magnetic resonance frequency up to GHz band. Coil as part of inductor was fabricated by lift off process. The dimension of RF inductor was designed 47un, rectangular shape, and measured properties. In the case of Ti/Ag air core type inductor shows Q of 9, inductance of 8.4 nH at 2 GHz. Magnetic film employed inductor shows inductance of 9 nH and FMR resonance frequency was 700 MHz.

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Isolation of Escherichia coli O157:H7 from animal feces and biochemical characteristics of Verotoxin-2 produced by these strains II. Purification and characterization of Verotoxin-2 Produced by Escherichia coli O157:H7 Isolated from animal feces (동물분변에서 Escherichia coli O157:H7의 분리 및 이들 균이 생산하는 Verotoxin-2의 생물화학적 특성 II. 동물분변에서 분리한 Escherichia coli O157:H7으로부터 Verotoxin-2의 정제 및 특성)

  • Cha, In-ho;Kim, Yong-hwan
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.379-387
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    • 1996
  • The objects of the present study were to establish the method of purification, subunit dissociation of verotoxin-2 (VT2) produced by Escherichia coli O157:H7, and to investigate the characteristics of purified verotoxin-2 such as molecular weight and composition of amino acid. The results were summerized as follows; Verotoxin-2 was extracted by addition of polymyxin B sulfate into bacterial cell lysate prepared from Escherichia coli O157:H7(KSC109). As an initial step, the bacterial cell lysate was precipitated with 30% saturated ammonium sulfate. The precipitated crude toxin was then subjected to anion-exchange, chromatofocusing and cation-exchange chromatography. Using this scheme, we obtained highly purified toxin with a specific activity of $1.1{\times}10^9$ $CD_{50}/mg$. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) for purified VT2 showed two protein bands. The upper band, approximately 32 Kd, was supposed as A subunit and the lower band, approximately 7.7 Kd, was supposed as B subunit. When the toxin was separated in the subunit-dissociating solution, two peaks emerged with retention times of 15 and 28 min by HPLC. These peaks represented A subunit and B subunit, respectively. The amino acid composition of purified VT2 were made up in order of glutamic acid, histamine, asparaginic acid, histidine, lysine, alanine and leucine etc. The largest amount among the amino acid composing VT2 was methionine.

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A Recombinant Human GM-CSF Protein Expressed as an Inclusion form in Escherichia coil Stimulates Colony Formation and Cell Proliferation in vitro

  • Lee, Ah-Young;Kim, Jin-Kyoo;Chung, Hye-Kyung;Bae, Eun-Kong;Hwang, Hung-Suk;Han, Jae-Yong;Lee, Choon-Taek;Hong, Soon-Kwnag;Youn, Hyun-Joo
    • Journal of Microbiology
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    • v.40 no.1
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    • pp.77-81
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    • 2002
  • Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic hematspoietic growth factor involved in the development of myeloid cells from bone marrow, and an activator of mature myeloid cells functioning in a variety of antimicrobial and inflammatory responses. Recently, recombinant GM-CSF is increasingly under clinical study for treatment of various diseases including cancer, infectious diseases and hematopoietic diseases as well as for an immune response modulator, In this study, we constructed a recombinant human GM-CSF (rhGM-CSF) expression plasmid with a pelB leader sequence and His. Tag under T7 promoter control. The expression construct was shown to produce a recombinant protein of 20 kDa in the 8M urea preparation, indicating the rhGM-CSF may be expressed as an insoluble inclusion form. The 20 kDa recombinant protein in 8M urea was transformed into the water-so1ub1e form by dialysis against PBS buffer (phosphate buffered saline). The soluble rhGM-CSF protein was shown to stimulate colony formation and cell proliferation in vitro, indicating that the rhGM-CSF could be refolded into its native form to show colony stimulating activity.

Photo-decomposition Characteristics of 2,4,6-Trinitrotoluene in a UV/$H_2O_2$ Process (2,4,6-Trinitrotoluene (TNT)의 광분해 특성)

  • Kwon, Bum-Gun;Choi, Won-Yong;Yoon, Je-Yong
    • Journal of Korean Society of Water and Wastewater
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    • v.24 no.6
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    • pp.775-788
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    • 2010
  • The decomposition of 2,4,6-trinitrotoluene (TNT) and the mass balance of nitrogen (N) species as products were investigated in a UV/H2O2system by varying pH, concentrations of $H_2O_2$, and $O_2$. All experiments were conducted in a semi-batch system employing a 50 mL reaction vessel and a coil-type quartz-tube reactor. In contrast with previous studies employing batch mode, TNT decomposition in the semi-batch mode was proportionally enhanced by increasing $H_2O_2$ concentration to 10 mM (0.034%), indicatingthat an inhibitory effect of excess $H_2O_2$on hydroxyl radical (${\cdot}OH$) can be negligible. N compounds are released as $NO_2^-$ in the early stages of the reaction, but $NO_2^-$ is rapidly oxidized to $NO_3^-$ by means of ${\cdot}OH$. $NH_4^+$ was also detected in this study and showed gradually the increase with increasing reaction time. In this study, $NH_4^+$ production can involve the reduction of nitro group of TNT concurrent with the production of $NO_3^-$. Of the N species originating from TNT decomposition, 12 ~ 72% were inorganic forms (i.e. [$NO_3^-$] + [$NO_2^-$] + [$NH_4^+$]). This result suggests that the large remaining N portions indicate that unidentified N compounds can exist.

Study on the control of VCM and its application to the vibration isolator (VCM의 제어 및 제진 장치 응용에 관한 연구)

  • Kim, Jin-Man;Nam, Taek-Kun
    • Journal of Advanced Marine Engineering and Technology
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    • v.37 no.1
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    • pp.91-98
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    • 2013
  • The degradation of durability and increase of fatigue on the ship are mainly caused by vibration of the engine and rotating machineries. The damper to minimize the influence from vibration is usually attached between the machineries and its base. General damper applied on the vessel is passive damper which is designed to attenuate specified frequency signals, i.e, high frequency vibration signals. But it is hard to anticipate its performance for low frequency signals. In this research, active vibration isolator using VCM is developed to suppress wide band vibration signals. Routh-Huritz's stable condition, ultimate sensitivity method and parameter tuning are applied to derive PID parameters and 2 and 4 phase choppers are also adapted to drive VCM. Simulation and experiments are executed to confirm the effectiveness of the proposed control schemes.