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  1. Article ; Online: Comprehensive analysis of multiple cytokines to stratify uropathogenic Escherichia coli pathogenesis in mouse model of urinary tract infection.

    Koley, Snehashis / Mukherjee, Mandira

    Cytokine

    2024  Volume 178, Page(s) 156577

    Abstract: Purpose: Urinary tract infection (UTI) is one of the most common human bacterial infections primarily caused by uropathogenic E. coli (UPEC). Empiric treatment in UTI cause emergence of multidrug resistance and limit treatment options. Understanding UTI ...

    Abstract Purpose: Urinary tract infection (UTI) is one of the most common human bacterial infections primarily caused by uropathogenic E. coli (UPEC). Empiric treatment in UTI cause emergence of multidrug resistance and limit treatment options. Understanding UTI at the molecular level with respect to the causative pathogen as well as subsequent host response pose an absolute necessity towards appropriate clinical management. This study aimed to investigate host cytokine response in mouse UTI model with respect to bacterial colonization and associated virulence gene expression upon infection.
    Method: Mouse UTI model was established with two clinical UPEC isolates E. coli NP105 and E. coli P025. UPEC colonization in bladder and kidney was evaluated by bacterial culture (CFU/ml). Histopathology of the tissues were examined by hematoxylin and eosin staining. PCR and real time PCR were used to detect the incidence and expression of respective bacterial genes. Cytokine concentrations in tissues and sera were evaluated using ELISA. GraphPad prism version 8.0.2 was used for statistical interpretation.
    Result: Highest bacterial colonization was observed on 7th and 9th day post infection (p.i). in bladder and kidney of mouse infected with E. coli P025 and E. coli NP105 respectively with a distinct difference in relative expression of fimH and papC adhesin genes in vivo. IL-1β level in tissues and sera of E. coli NP105 and E. coli P025 infected mouse was significantly different but the IL-17A, GCSF, TGF-β levels were comparable.
    Conclusion: These findings show a role of IL1β to stratify pathogenicity of UPEC in mouse UTI model.
    MeSH term(s) Humans ; Animals ; Mice ; Uropathogenic Escherichia coli ; Cytokines ; Escherichia coli Infections/microbiology ; Urinary Tract Infections/microbiology ; Urinary Bladder/microbiology
    Chemical Substances Cytokines
    Language English
    Publishing date 2024-03-12
    Publishing country England
    Document type Journal Article
    ZDB-ID 1018055-2
    ISSN 1096-0023 ; 1043-4666
    ISSN (online) 1096-0023
    ISSN 1043-4666
    DOI 10.1016/j.cyto.2024.156577
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    Zs.A 2840: Show issues Location:
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  2. Article ; Online: Type 1 fimbrial phase variation in multidrug-resistant asymptomatic uropathogenic Escherichia coli clinical isolates upon adherence to HTB-4 cells.

    Ghosh, Arunita / Mukherjee, Mandira

    Folia microbiologica

    2024  

    Abstract: The adherence of bladder uroepithelial cells, subsequent expression, and regulation of type 1 fimbrial genes (key mediator of attachment) in clinical multidrug-resistant uropathogenic Escherichia coli (MDR-UPECs) isolated from individuals with ... ...

    Abstract The adherence of bladder uroepithelial cells, subsequent expression, and regulation of type 1 fimbrial genes (key mediator of attachment) in clinical multidrug-resistant uropathogenic Escherichia coli (MDR-UPECs) isolated from individuals with asymptomatic bacteriuria (ABU) remain unexplored till date. Therefore, this study aimed to investigate the underlying molecular mechanisms associated with the adherence of clinical MDR-ABU-UPECs to human a uroepithelial cell line (HTB-4), both in the absence and presence of D-Mannose. These investigations focused on phase variation, expression, and regulation of type 1 fimbriae and were compared to a prototype ABU-strain (E. coli 83972) and symptomatic MDR-UPECs. Discordant to the ABU prototype strain, MDR-ABU-UPECs exhibited remarkable adhesive capacity that was significantly reduced after D-mannose exposure, fairly like the MDR symptomatic UPECs. The type 1 fimbrial phase variation, determined by the fim switch analysis, asserted the statistically significant incidence of "both OFF and ON" orientation among the adherent MDR-ABU-UPECs with a significant reduction in phase-ON colonies post-D-mannose exposure, akin to the symptomatic ones. This was indicative of an operative and alternating type 1 fimbrial phase switch. The q-PCR assay revealed a coordinated action of the regulatory factors; H-NS, IHF, and Lrp on the expression of FimB and FimE recombinases, which further controlled the function of fimH and fimA genes in ABU-UPECs, similar to symptomatic strains. Therefore, this study is the first of its kind to provide an insight into the regulatory crosstalk of different cellular factors guiding the adhesion of ABU-UPECs to the host. Additionally, it also advocated for the need to accurately characterize ABU-UPECs.
    Language English
    Publishing date 2024-04-03
    Publishing country United States
    Document type Journal Article
    ZDB-ID 240503-9
    ISSN 1874-9356 ; 0015-5632
    ISSN (online) 1874-9356
    ISSN 0015-5632
    DOI 10.1007/s12223-024-01159-y
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  3. Article ; Online: Cytokine profile in HTB-4 cells with respect to

    Koley, Snehashis / Mukherjee, Mandira

    Future microbiology

    2023  Volume 18, Page(s) 489–503

    Abstract: Background: ...

    Abstract Background:
    MeSH term(s) Humans ; Urinary Tract Infections/drug therapy ; Uropathogenic Escherichia coli/genetics ; Anti-Bacterial Agents/therapeutic use ; Virulence Factors/genetics ; Real-Time Polymerase Chain Reaction ; Escherichia coli Infections/drug therapy ; Adhesins, Escherichia coli/genetics ; Fimbriae Proteins/genetics
    Chemical Substances 4-trifluoromethylsalicylic acid (328-90-5) ; Anti-Bacterial Agents ; Virulence Factors ; fimH protein, E coli ; Adhesins, Escherichia coli ; Fimbriae Proteins (147680-16-8)
    Language English
    Publishing date 2023-06-08
    Publishing country England
    Document type Journal Article
    ZDB-ID 2254620-0
    ISSN 1746-0921 ; 1746-0913
    ISSN (online) 1746-0921
    ISSN 1746-0913
    DOI 10.2217/fmb-2022-0240
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  4. Article ; Online: Occurrence of Imipenem-Resistant Uropathogenic Escherichia coli in Pregnant Women: An Insight into Their Virulence Profile and Clonal Structure.

    Koley, Snehashis / Ghosh, Arunita / Mukherjee, Mandira

    Current microbiology

    2024  Volume 81, Issue 2, Page(s) 56

    Abstract: Uropathogenic Escherichia coli (UPEC) is the predominant pathogen in Urinary Tract Infection (UTI) in pregnant and non-pregnant women. Limited studies were initiated to explore UPEC from pregnant women with respect to imipenem resistance, pathogenicity, ... ...

    Abstract Uropathogenic Escherichia coli (UPEC) is the predominant pathogen in Urinary Tract Infection (UTI) in pregnant and non-pregnant women. Limited studies were initiated to explore UPEC from pregnant women with respect to imipenem resistance, pathogenicity, and their clonal lineage. In this study, imipenem resistance, phylogenetic background, virulence-associated genes, and clonal characteristics in UPECs isolated from pregnant and non-pregnant cohorts were investigated. E. coli was identified biochemically from urine culture-positive samples from pregnant and non-pregnant women. Carbapenem (meropenem, ertapenem, imipenem) susceptibility was determined by Kirby-Bauer disk diffusion test. The pathogenic determinants were identified by PCR. MEGA 11 was used to interpret clonal lineages from MLST. GraphPad Prism 8.0 and SPSS 26.0 were used for statistical interpretation. Results indicated highest resistance against imipenem compared to meropenem and ertapenem in UPECs isolated from pregnant (UPECp; 63.89%) and non-pregnant (UPECnp; 87.88%) women. Although phylogroup E was predominant in both imipenem-resistant isolates, acquisition of virulence factors was higher among UPECnp than UPECp. Akin to this observation, the presence of PAI III
    MeSH term(s) Pregnancy ; Humans ; Female ; Male ; Imipenem/pharmacology ; Virulence/genetics ; Uropathogenic Escherichia coli/genetics ; Ertapenem/pharmacology ; Meropenem ; Multilocus Sequence Typing ; Phylogeny ; Pregnant Women ; Virulence Factors/genetics
    Chemical Substances Imipenem (71OTZ9ZE0A) ; Ertapenem (G32F6EID2H) ; Meropenem (FV9J3JU8B1) ; Virulence Factors
    Language English
    Publishing date 2024-01-09
    Publishing country United States
    Document type Journal Article
    ZDB-ID 134238-1
    ISSN 1432-0991 ; 0343-8651
    ISSN (online) 1432-0991
    ISSN 0343-8651
    DOI 10.1007/s00284-023-03576-7
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    Zs.A 1446: Show issues Location:
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  5. Article ; Online: Combination of bactericidal antibiotics and inhibitors of Universal stress protein A (UspA): a potential therapeutic alternative against multidrug resistant Escherichia coli in urinary tract infections.

    Bandyopadhyay, Debojyoty / Mukherjee, Mandira

    The Journal of antibiotics

    2021  Volume 75, Issue 1, Page(s) 21–28

    Abstract: The increasing incidence of multidrug resistant uropathogenic E. coli (MDR-UPEC), the most common opportunistic pathogen in urinary tract infections (UTI) pose a global health problem and demands searching for alternative therapeutics. Antibiotics ... ...

    Abstract The increasing incidence of multidrug resistant uropathogenic E. coli (MDR-UPEC), the most common opportunistic pathogen in urinary tract infections (UTI) pose a global health problem and demands searching for alternative therapeutics. Antibiotics generate oxidative stress in bacteria which results in overexpression of the universal stress protein, UspA that helps in bacterial survival. An in silico study showed that two compounds ZINC000104153710, and ZINC000000217308 effectively bound bacterial UspA. This study aimed to determine the activity of ZINC000104153710, and ZINC000000217308 against bacterial UspA function in MDR-UPEC in vitro. Twenty-five highly MDR-UPEC were screened against ZINC000104153710, and, ZINC000000217308 either alone or in combination with the bactericidal antibiotics; ciprofloxacin (CIP), ceftazidime(CAZ), gentamicin(GEN) respectively by determining minimum inhibitory concentrations (MICs) using a broth microdilution assay. Additionally, the effect of ZINC000104153710, and ZINC000000217308 in the absence and presence of antibiotics on the bacteria was monitored by bacterial growth curve assays, ROS production, structure of the organism by scanning electron microscopy (FESEM) and quantitating UspA using a western blot technique. A 2-8 fold reduction in MIC values against ZINC000104153710, and ZINC000000217308 was observed against all 25 MDR-UPEC isolates in the presence of antibiotics with no alteration in intracellular ROS production. Discrete changes in cell morphology was evident in bacteria treated with ZINC000104153710 or ZINC000000217308 and antibiotics individually by FESEM compared with untreated control. Reduction in the level of UspA protein in bacteria treated with combination of ZINC000104153710 or ZINC000000217308 with individual antibiotics established their ability to inhibit UspA whose expression was elevated in presence of antibiotics alone. Therefore this study validated ZINC000104153710, and ZINC000000217308 as potent inhibitors of bacterial UspA function and indicated their potential as alternative therapeutics to combat the MDR-UPEC.
    MeSH term(s) Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/antagonists & inhibitors ; Drug Resistance, Multiple, Bacterial/drug effects ; Escherichia coli/drug effects ; Escherichia coli/genetics ; Escherichia coli Infections/drug therapy ; Escherichia coli Infections/microbiology ; Escherichia coli Infections/urine ; Heat-Shock Proteins/antagonists & inhibitors ; Humans ; Microbial Sensitivity Tests ; Reactive Oxygen Species ; Urinary Tract Infections/drug therapy ; Urinary Tract Infections/microbiology ; Urinary Tract Infections/urine
    Chemical Substances Anti-Bacterial Agents ; Bacterial Proteins ; Heat-Shock Proteins ; Reactive Oxygen Species ; universal stress protein A, Bacteria
    Language English
    Publishing date 2021-09-15
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 390800-8
    ISSN 1881-1469 ; 0021-8820 ; 0368-3532
    ISSN (online) 1881-1469
    ISSN 0021-8820 ; 0368-3532
    DOI 10.1038/s41429-021-00477-4
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    Zs.A 207: Show issues Location:
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  6. Article ; Online: Reactive oxygen species and uspA overexpession: an alternative bacterial response toward selection and maintenance of multidrug resistance in clinical isolates of uropathogenic E. coli.

    Bandyopadhyay, Debojyoty / Mukherjee, Mandira

    European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology

    2020  Volume 39, Issue 9, Page(s) 1753–1760

    Abstract: Emergence of multidrug resistance (MDR) in uropathogenic E. coli (UPEC) demands alternative therapeutic interventions. Bactericidal antibiotics at their sub-inhibitory concentration stimulate production of reactive oxygen species (ROS) that results in ... ...

    Abstract Emergence of multidrug resistance (MDR) in uropathogenic E. coli (UPEC) demands alternative therapeutic interventions. Bactericidal antibiotics at their sub-inhibitory concentration stimulate production of reactive oxygen species (ROS) that results in oxidative stress, generates mutations, and alters transcription of different genes. Sub-inhibitory concentration of antibiotics facilitates selection of highly resistant population. Universal stress protein A (uspA) overexpression in MDR-UPEC at sub-inhibitory bactericidal antibiotics concentration was investigated to explore alternative survival strategy against them. Fifty clinical UPEC isolates were screened. Minimum inhibitory concentration (MIC) against three different bactericidal antibiotics (ciprofloxacin, CIP; ceftazidime, CAZ; gentamycin, GEN) was determined by broth dilution method; ROS production by DCFDA and overexpression of uspA by real-time PCR were determined at the sub-inhibitory concentration of antibiotics. DNA ladder formation and SEM studies were performed with drug untreated and treated samples. Statistical analysis was done by Student's t test and Pearson's correlation analysis; 25 out of 50 UPEC exhibited high MIC against CIP (> 200 μg/ml), CAZ (> 500 μg/ml), GEN (> 500 μg/ml), with varied ROS production (p ≤ 0.001) in treated than untreated controls. DNA ladder formation confirmed ROS production in drug-treated samples. SEM analysis revealed unaltered cell morphology in both untreated and drug-treated bacteria. uspA was universally overexpressed in all 25 UPEC. A significant correlation (p ≤ 0.001) between ROS production and uspA overexpression was observed in 19 out of 25 MDR isolates at sub-inhibitory doses of the bactericidal antibiotics. Therefore, this study highlights an alternative strategy that the MDR isolates may acquire when exposed to sub-inhibitory drug concentration for their survival.
    MeSH term(s) Anti-Bacterial Agents/pharmacology ; Drug Resistance, Multiple, Bacterial ; Escherichia coli Infections/drug therapy ; Escherichia coli Infections/microbiology ; Humans ; Microbial Sensitivity Tests ; Reactive Oxygen Species/metabolism ; Staphylococcal Protein A/metabolism ; Urinary Tract Infections/drug therapy ; Urinary Tract Infections/microbiology ; Uropathogenic Escherichia coli/drug effects ; Uropathogenic Escherichia coli/metabolism ; Uropathogenic Escherichia coli/ultrastructure
    Chemical Substances Anti-Bacterial Agents ; Reactive Oxygen Species ; Staphylococcal Protein A
    Language English
    Publishing date 2020-05-12
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 603155-9
    ISSN 1435-4373 ; 0934-9723 ; 0722-2211
    ISSN (online) 1435-4373
    ISSN 0934-9723 ; 0722-2211
    DOI 10.1007/s10096-020-03903-x
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  7. Article: Impact of Mutation on the Structural Stability and the Conformational Landscape of Inhibitor-Resistant TEM β-Lactamase: A High-Performance Molecular Dynamics Simulation Study

    Mukherjee, Sandip K. / Mukherjee, Mandira / Mishra, Padmaja P.

    Journal of physical chemistry. 2021 Oct. 05, v. 125, no. 40

    2021  

    Abstract: Gain-of-function mutations and structural adjustment toward β-lactamase inhibitors in the TEM-type β-lactamases among the uropathogenic E. coli (UPEC) culminate in treatment complications and demands detailed investigation. In this study, uncharacterized ...

    Abstract Gain-of-function mutations and structural adjustment toward β-lactamase inhibitors in the TEM-type β-lactamases among the uropathogenic E. coli (UPEC) culminate in treatment complications and demands detailed investigation. In this study, uncharacterized amino acid substitutions, M69L/I84V/W165G/V184A/V262I/N276S, in inhibitor-resistant TEM (IRT) β-lactamase isolated from clinical UPEC were subjected to extensive molecular dynamics (EMD) simulations for 100 ns to estimate parameters such as root-mean-square deviation (RMSD), root-mean-square fluctuation (RMSF), the radius of gyration (Rg), contour plot (Rg/RMSD), secondary structure element (SSE), etc. Residue interaction networks, principal component analysis (PCA), and correlation heatmaps were generated to predict the relation between functionally important atomic motions to uncover the structural stability of the mutants. To avoid the false positive conclusion of the simulation study, we performed three identically parameterize replicas of 100 ns each. Alterations in hydrophobic interactions resulted in conformation changes exhibited as comparable residue interaction networks. Besides, PCA and porcupine plot analysis based on the ensemble of structure from molecular dynamics trajectories revealed the collective atomic motions of the IRT variants that impart structural flexibility to their active site loop. This study conducted on IRT mutants that delineate intricate protein motions contributes to their stability and folding, which is an absolute necessity for designing candidate molecules owing to the clinical threat of emerging resistance against potent β-lactam antibiotics.
    Keywords active sites ; amino acids ; gain-of-function mutation ; hydrophobicity ; molecular dynamics ; principal component analysis ; uropathogenic Escherichia coli
    Language English
    Dates of publication 2021-1005
    Size p. 11188-11196.
    Publishing place American Chemical Society
    Document type Article
    ISSN 1520-5207
    DOI 10.1021/acs.jpcb.1c05988
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  8. Article: Uropathogenic Escherichia coli in India—an Overview on Recent Research Advancements and Trends

    Ghosh, Arunita / Bandyopadhyay, Debojyoty / Koley, Snehashis / Mukherjee, Mandira

    Applied biochemistry and biotechnology. 2021 July, v. 193, no. 7

    2021  

    Abstract: Urinary tract infection (UTI), a prevalent disease in India, also ranks among the most common infections in developing countries. The rapid emergence of antibiotic-resistant uropathogenic Escherichia coli (UPECs), the leading etiologic agent of UTI, in ... ...

    Abstract Urinary tract infection (UTI), a prevalent disease in India, also ranks among the most common infections in developing countries. The rapid emergence of antibiotic-resistant uropathogenic Escherichia coli (UPECs), the leading etiologic agent of UTI, in the last few years, led to an upsurge in the health care cost. This caused a considerable economic burden, especially in low-middle income country, India. This review aimed to provide an explicit overview of the recent advancements in E. coli–mediated UTI in India by incorporation of valuable information from the works published in PubMed and Google Scholar in the last six years (2015 to August, 2020). The literature survey demonstrated UPECs as the most predominant uropathogen in India, especially among females, causing both asymptomatic bacteriuria (ABU) and symptomatic UTI. An overall increasing national trend in resistance to penicillins, cephalosporins, aminoglycosides, fluoroquinolones, and sulfonamides was perceived irrespective of ABU and symptomatic UPECs during the aforementioned study period. High incidences of multidrug resistance, extended-spectrum β-lactamases, metallo β-lactamases, and AmpCs in UPECs were reported. Notable information on the pathogenic profiles, phylogroups, pathogenicity islands, and evidence of pathoadaptive FimH mutations was described. Alternative therapeutics and potential drug targets against UPECs were also reconnoitered. Therefore, the nationwide widespread occurrences of highly virulent MDR UPEC together with the limited availability of therapeutics highlighted the urgent need for promotion and invention of alternative therapeutics, search for which had already been started. Moreover, investigation of several mechanisms of UPEC infection and the search for potential drug targets might help to design newer therapeutics.
    Keywords aminoglycosides ; antibiotic resistance ; biotechnology ; cephalosporins ; etiological agents ; fluoroquinolones ; health care costs ; income ; multiple drug resistance ; surveys ; therapeutics ; urinary tract diseases ; uropathogenic Escherichia coli ; virulence ; India
    Language English
    Dates of publication 2021-07
    Size p. 2267-2296.
    Publishing place Springer US
    Document type Article
    Note Review
    ZDB-ID 392344-7
    ISSN 0273-2289
    ISSN 0273-2289
    DOI 10.1007/s12010-021-03521-z
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  9. Article: Characterization and Bio-Typing of Multidrug Resistance Plasmids From Uropathogenic

    Mukherjee, Sandip Kumar / Mukherjee, Mandira

    Frontiers in microbiology

    2019  Volume 10, Page(s) 2913

    Abstract: Urinary tract infection is primarily caused ... ...

    Abstract Urinary tract infection is primarily caused by
    Language English
    Publishing date 2019-12-18
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2587354-4
    ISSN 1664-302X
    ISSN 1664-302X
    DOI 10.3389/fmicb.2019.02913
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  10. Article ; Online: Conjugal transfer of PMQR from uropathogenic E.coli under high ciprofloxacin selection pressure generates gyrA mutation.

    Basu, Shreya / Mukherjee, Mandira

    Microbial pathogenesis

    2019  Volume 132, Page(s) 26–29

    Abstract: Incidence of high fluoroquinolone resistance has been rising rapidly worldwide. Resistance against fluoroquinolones can be either chromosomal or plasmid mediated. Plasmid mediated quinolone resistant(PMQR) genes impart low level of resistance against ... ...

    Abstract Incidence of high fluoroquinolone resistance has been rising rapidly worldwide. Resistance against fluoroquinolones can be either chromosomal or plasmid mediated. Plasmid mediated quinolone resistant(PMQR) genes impart low level of resistance against fluoroquinolones but provides favorable background for selection of additional chromosomally encoded resistance mechanisms. In the natural habitat, conjugal transfer of PMQR genes provides a vehicle for dissemination of fluoroquinolone resistance. Our study indicated successful transmission of PMQR determinants(aac(6')-Ib-cr, qnrA, qnrB, qnrS, oqxB) from ciprofloxacin resistant clinical uropathogenic E.coli(UPEC) isolates to susceptible E.coliJ53Azide-resistant strain in vitro in presence of high ciprofloxacin (5 μg/ml) selection pressure generating transconjugants that exhibited varied MIC(25-800μg/ml) towards the drug with acquired mutations Ser83Leu and Asp87Asn in the quinolone resistant determining regions(QRDR) in gyrA. Therefore this is a first study of its kind that identified high rate of gyrA mutations among transconjugants selected under high ciprofloxacin pressure resulting from bacterial conjugation a common phenomenon in natural habitat along with PMQR gene transmission which imposes a major public health concern.
    MeSH term(s) Anti-Bacterial Agents/pharmacology ; Ciprofloxacin/pharmacology ; Conjugation, Genetic ; DNA Gyrase/genetics ; DNA, Bacterial/genetics ; Drug Resistance, Bacterial/drug effects ; Drug Resistance, Bacterial/genetics ; Fluoroquinolones ; Genes, Bacterial/genetics ; Microbial Sensitivity Tests ; Mutation ; Plasmids/genetics ; Quinolones ; Recombination, Genetic ; Uropathogenic Escherichia coli/drug effects ; Uropathogenic Escherichia coli/genetics
    Chemical Substances Anti-Bacterial Agents ; DNA, Bacterial ; Fluoroquinolones ; Quinolones ; Ciprofloxacin (5E8K9I0O4U) ; DNA Gyrase (EC 5.99.1.3)
    Language English
    Publishing date 2019-04-16
    Publishing country England
    Document type Journal Article
    ZDB-ID 632772-2
    ISSN 1096-1208 ; 0882-4010
    ISSN (online) 1096-1208
    ISSN 0882-4010
    DOI 10.1016/j.micpath.2019.04.021
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