• Urogenital Tract Infection
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• 제13권3호
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• pp.58-65
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• 2018

Purpose: This study aimed to investigate the relationship between uropathogens of infants with febrile urinary tract infection (UTI) and vesicoureteral reflux (VUR). Materials and Methods: We analyzed 308 infants hospitalized for febrile UTI between January 2010 and December 2015, and assessed the voiding cystourethrography (VCUG). The medical records, including clinical symptoms, laboratory findings, urinalysis, urine culture tests, ultrasound (US), dimercaptosuccinic acid scan, and VCUG, were retrospectively obtained. The incidences of VUR and high-grade VURs (III, IV, and V) were analyzed in 4 groups categorized by uropathogens and renal US findings. Results: The mean age of 308 infants was $3.29{\pm}2.18months$. The male-to-female ratio was 3.46:1. In urine culture tests, 267 infants (86.69%) showed single bacterial uropathogen; Escherichia coli in 241 infants (78.25%) and non-E. coli uropathogens in 26 infants (8.44%). Multiple distinctive microorganisms were identified as causative uropathogens in 41 infants (13.31%). Abnormal findings of US and VCUG were identified in 216 and 64 patients, respectively. In 308 infants, the incidences of VUR and high-grade VUR were not different among the 4 groups. In 239 male infants, the incidences of high-grade VUR were higher in patients with non-E. coli single or multiple uropathogen and with abnormal US findings (p=0.042). Conclusions: In male infants with non-E. coli uropathogen or multiple uropathogens and with abnormal US findings at febrile UTI, there was an increased chance of finding high-grade VURs on subsequent VCUG tests.

• Journal of Microbiology and Biotechnology
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• 제25권1호
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• pp.137-142
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• 2015

Trimethoprim-sulfamethoxazole (TMP-SMX) has been used for the treatment of urinary tract infections, but increasing resistance to TMP-SMX has been reported. In this study, we analyzed TMP-SMX resistance genes and their relatedness with integrons and insertion sequence common regions (ISCRs) in uropathogenic gram-negative bacilli. Consecutive nonduplicate TMP-SMX nonsusceptible clinical isolates of E. coli, K. pneumoniae, Acinetobacter spp., and P. aeruginosa were collected from urine. The minimal inhibitory concentration was determined by Etest. TMP-SMX resistance genes (sul and dfr), integrons, and ISCRs were analyzed by PCR and sequencing. A total of 45 E. coli (37.8%), 15 K. pneumoniae (18.5%), 12 Acinetobacter spp. (70.6%), and 9 Pseudomonas aeruginosa (30.0%) isolates were found to be resistant to TMP-SMX. Their MICs were all over 640. In E. coli and K. pneumoniae, sul1 and dfr genes were highly prevalent in relation with integron1. The sul3 gene was detected in E. coli. However, in P. aeruginosa and Acinetobacter spp., only sul1 was prevalent in relation with class 1 integron; however, dfr was not detected and sul2 was less prevalent than in Enterobacteriaceae. ISCR1 and/or ISCR2 were detected in E. coli, K. pneumoniae, and Acinetobacter spp. but the relatedness with TMP-SMX resistance genes was not prominent. ISCR14 was detected in six isolates of E. coli. In conclusion, resistance mechanisms for TMP-SMX were different between Enterobacteriaceae and glucose non-fermenting gram-negative bacilli. Class 1 integron was widely disseminated in uropathogenic gram-negative baciili, so the adoption of prudent use of antimicrobial agents and the establishment of a surveillance system are needed.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 1999년도 학술발표회 진행표 및 논문초록
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• pp.43-43
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• 1999

Signal peptides of secretary proteins interact with various membranes and non-membrane components during the translocation. We investigated the interaction of signal peptides of ribose binding protein (RBP) with Escherichia coli (E.coli) signal recognition particle (SRP), SecA and lipid bilayer. Previous studies showed that the functional signal peptides inhibit the GTPase activity of E.coli SRP which consisted of F로 and 4.5S RNA.(omitted)

• 대한수의학회지
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• 제50권3호
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• pp.231-237
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• 2010

This study was conducted to investigate characteristics and antimicrobial susceptibility of Escherichia (E.) coli isolates isolated from vaginal mucosa and clitorial fossa of 105 Thoroughbred mares suspicious of the genital disease in Korea during the period from March 2006 to July 2007. Ninety six E. coli isolates were identified as standard biochemical properties and using BIOLOG system. Fifty three isolates (55.2%) could be classified into a total of 21 O serotypes and forty three isolates (44.8%) were non-typeable with 51 O antisera used in this study. The verotoxin 1 (VT 1) and verotoxin 2 genes were analyzed by multiplex PCR. Among them, one isolate was detected VT 1 gene (130 bp). Most of isolates showed a high susceptibility in ciprofloxacin (100%), enrofloxacin (100%), norfloxacin (100%), cefoxitin (96.9%), gentamicin (96.9%), sulphamethoxazole (96.9%), nitrofurantoin (94.8%), amikacin (93.8%), nalidixic acid (92.7%) and tetracycline (90.6%). These results may provide the basic information to establish strategies for the treatment and prevention of reproductive disease in Thoroughbred mares in Korea.

• Journal of Microbiology and Biotechnology
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• 제34권5호
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• pp.1126-1134
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• 2024

The production of disulfide bond-containing recombinant proteins in Escherichia coli has traditionally been done by either refolding from inclusion bodies or by targeting the protein to the periplasm. However, both approaches have limitations. Two broad strategies were developed to allow the production of proteins with disulfide bonds in the cytoplasm of E. coli: i) engineered strains with deletions in the disulfide reduction pathways, e.g. SHuffle, and ii) the co-expression of oxidative folding catalysts, e.g. CyDisCo. However, to our knowledge, the relative effectiveness of these strategies has not been properly evaluated. Here, we systematically compare the purified yields of 14 different proteins of interest (POI) that contain disulfide bonds in their native state when expressed in both systems. We also compared the effects of different background strains, commonly used promoters, and two media types: defined and rich autoinduction. In rich autoinduction media, POI which can be produced in a soluble (non-native) state without a system for disulfide bond formation were produced in higher purified yields from SHuffle, whereas all other proteins were produced in higher purified yields using CyDisCo. In chemically defined media, purified yields were at least 10x higher in all cases using CyDisCo. In addition, the quality of the three POI tested was superior when produced using CyDisCo.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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• pp.141-145
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• 2001

Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all carotenoids. IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of l-deoxy-D-xylulose-5-phosphate (DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoisomerase and encoded by dxr. To determine if one or more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains (DH5$\alpha$, XL1-Blue, and JMl0l) that had been engineered to produce lycopene. Lycopene production was improved significantly in strains transformed with the dxs expression vectors. When the dxs gene was expressed from the arabinose-inducible araBAD promoter ( $P_{BAD}$) on a medium-copy plasmid, lycopene production was 2-fold higher than when dxs was expressed from the IPTG-inducible trc and lac promoters ( $P_{trc}$ and $P_{lac}$, respectively) on medium-copy and high-copy plasmids. Given the low final densities of cells expressing dxs from IPTG-inducible promoters, the low lycopene production was probably due to the metabolic burden of plasmid maintenance and an excessive drain of central metabolic intermediates. At arabinose concentrations between 0 and 1.33 roM, cells expressing both dxs and dxr from $P_{BAD}$ on a medium-copy plasmid produced 1.4 - 2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene . production in cells expressing both dxs and dxr was lower than in cells expressing dxs only. A comparison of the three E. coli strains transformed with the arabinose-inducible dxs on a medium-copy plasmid revealed that lycopene production was highest in XLI-Blue.LI-Blue.

• 상하수도학회지
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• 제21권2호
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• pp.235-242
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• 2007

Road runoff water includes various heavy metals (zinc, Zn; lead, Pb; copper, Cu; chrome, Cr; cadmium, Cd; etc.) and pathogens (E-coli and coliform). Since these pollutants are significantly harmful to human beings and have negative impact on water streams, numerous studies have been conducted to determine the characterization of these non-point pollutants from road runoff water. However, since these non-point pollutant concentrations vary depending on road traffic, road construction, and road maintenance, measurement of pollutant loadings in different site is necessary to estimate the effect of road runoff water on drinking water source. The objective of this study was to examine the quality of road runoff water from a city bridge in Seoul, Korea. This study was conducted for two years to assess annual discharge pollution loads. In this study, five key heavy metals (Zn, Pb, Cu, Cr, and Cd) and two pathogens (E-coli and coliform) were measured at 18 different events. The pollutant load mass transported was always higher than the corresponding runoff volume for Zn, Cu, and Cd, while Pb and Cr showed similar values between the load mass transported and the corresponding runoff volume. The event mean concentrations were Zn (0.908 mg/L), Pb (0.092 mg/L), Cu (0.141 mg/L), Cr (0.023 mg/L), and Cd (0.006 mg/L). Like Zn, Cu, and Cd, E-coli and coliform values (relatively high in Summer and Fall) are higher at the beginning of each event and decrease afterwards.

• Journal of Microbiology and Biotechnology
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• 제31권5호
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• pp.710-716
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• 2021

A risk analysis of Shiga toxin (Stx)-encoding bacteriophage was carried out by confirming the transduction phage to non-Stx-producing Escherichia coli (STEC) and subsequent expression of the Shiga toxin genes. The virulence factor stx1 was identified in five phages, and both stx1 and stx2 were found in four phages from a total of 19 phage isolates with seven non-O157 STEC strains. The four phages, designated as ϕNOEC41, ϕNOEC46, ϕNOEC47, and ϕNOEC49, belonged morphologically to the Myoviridae family. The stabilities of these phages to temperature, pH, ethanol, and NaClO were high with some variabilities among the phages. The infection of five non-STEC strains by nine Stx-encoding phages occurred at a rate of approximately 40%. Non-STEC strains were transduced by Stx-encoding phage to become lysogenic strains, and seven convertant strains had stx1 and/or stx2 genes. Only the stx1 gene was transferred to the receptor strains without any deletion. Gene expression of a convertant having both stx1 and stx2 genes was confirmed to be up to 32 times higher for Stx1 in 6% NaCl osmotic media and twice for Stx2 in 4% NaCl media, compared with expression in low-salt environments. Therefore, a new risk might arise from the transfer of pathogenic genes from Stx-encoding phages to otherwise harmless hosts. Without adequate sterilization of food exposed to various environments, there is a possibility that the toxicity of the phages might increase.

• 대한수의학회지
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• 제47권1호
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• pp.67-75
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• 2007

Environmental mastitis has increased particularly in well-managed or low somatic cell countherds that have successfully controled contagious pathogens. Major pathogens of environmental mastitisare Escherichia coli (E. coli) and Streptococcus uberis. The present study was conducted to investigate1,865 quaters of 241 Korean dairy farms from 2001 to 2004. Prevalence of major gram-negative bacteriaisolated from mastitis milk were E. coli (22.7%) and Enterobacter spp. (16.3%) in coliforms and Pseudomoassp. (10.3%) and Serratia spp. (7.9%) in non-coliforms. The results on antibiotic susceptibility by agardifusion test against these pathogens were 86.7% in piperaciliin, 94.6% in cefepime, 85.5% in amikacin,87.7% in gentamicin and so on. In contrast, the susceptibility against ampicillin (41.9%), cephalothin (9.9%),streptomycin (39.9%) and tetracycline (46.7%) appeared to be below 50%. Gram-negative bacteria showed(96.8%). Acording to year, distribution of high $256{\sim}64{\mu}g/ml$ on cephalothin get increased, but the othersare diferent. These findings demonstrate that major gram-negative bacteria were E. coli and Enterobacterspp. isolates, and often encountered the diverse antibiotic resistant patterns.

• 한국미생물·생명공학회지
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• 제51권1호
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• pp.37-42
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• 2023

Neutral and non-essential amino acid, glutamine (Gln), plays an essential role in supplying nitrogen to all the amino acids and nucleotides in the mammalian body. Gln is also the most important carbon source that provides intermediates for gluconeogenesis and fatty acid synthesis and supplements the tricarboxylic acid cycle in fast-growing cancer cells. Among the known 14 Gln transporter genes, soluted carrier family 1 member 5 (SLC1A5) has been reported to be closely associated with cancer cell growth. Three variants (v1, v2, and v3) have been derived from SLC1A5. Here, we established a heterologous gene expression system for the active form of human SLC1A5 variant-2 (hSLC1A5v2) in Escherichia coli. v2 is the smallest variant that has not yet been studied. Four expression systems were investigated: pBAD, pCold, pET, and pQE. We also addressed the problem of codon usage bias. Although pCold and pET overexpressed hSLC1A5v2 in E. coli, they were functionally inactive. hSLC1A5v2 using the pBAD system was able to catalyze the successful transport of Gln, even if it was not highly expressed. Initial activity of hSLC1A5v2 for [¹⁴C] Gln uptake in E. coli reached up to 6.73 μmole·min^-1·gDW^-1 when the cell was induced with 80 mM L-arabinose. In this study, we demonstrated a heterologous expression system for the human membrane protein, SLC1A5, in E. coli. Our results can be used for the functional comparison of SLC1A5 variants (v1, v2, and v3) in future studies, to facilitae the developement of SLC1A5 inhibitors as effective anticancer drugs.