scholarly journals d-Mannose Treatment neither Affects Uropathogenic Escherichia coli Properties nor Induces Stable FimH Modifications

Molecules ◽  
2020 ◽  
Vol 25 (2) ◽  
pp. 316 ◽  
Author(s):  
Daniela Scribano ◽  
Meysam Sarshar ◽  
Carla Prezioso ◽  
Marco Lucarelli ◽  
Antonio Angeloni ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacterial Growth ◽  
Good Alternative ◽  
Uropathogenic Escherichia Coli ◽  
Bacterial Metabolism ◽  
Upec Strain ◽  
Bladder Cell ◽  
Microbiological Methods ◽  
Strain Cft073 ◽  
Tract Infections

Urinary tract infections (UTIs) are mainly caused by uropathogenic Escherichia coli (UPEC). Acute and recurrent UTIs are commonly treated with antibiotics, the efficacy of which is limited by the emergence of antibiotic resistant strains. The natural sugar d-mannose is considered as an alternative to antibiotics due to its ability to mask the bacterial adhesin FimH, thereby preventing its binding to urothelial cells. Despite its extensive use, the possibility that d-mannose exerts “antibiotic-like” activity by altering bacterial growth and metabolism or selecting FimH variants has not been investigated yet. To this aim, main bacterial features of the prototype UPEC strain CFT073 treated with d-mannose were analyzed by standard microbiological methods. FimH functionality was analyzed by yeast agglutination and human bladder cell adhesion assays. Our results indicate that high d-mannose concentrations have no effect on bacterial growth and do not interfere with the activity of different antibiotics. d-mannose ranked as the least preferred carbon source to support bacterial metabolism and growth, in comparison with d-glucose, d-fructose, and l-arabinose. Since small glucose amounts are physiologically detectable in urine, we can conclude that the presence of d-mannose is irrelevant for bacterial metabolism. Moreover, d-mannose removal after long-term exposure did not alter FimH’s capacity to bind to mannosylated proteins. Overall, our data indicate that d-mannose is a good alternative in the prevention and treatment of UPEC-related UTIs.

scholarly journals Role of Motility in the Colonization of Uropathogenic Escherichia coli in the Urinary Tract

2005 ◽  
Vol 73 (11) ◽  
pp. 7644-7656 ◽  
Author(s):  
M. Chelsea Lane ◽  
Virginia Lockatell ◽  
Greta Monterosso ◽  
Daniel Lamphier ◽  
Julia Weinert ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Wild Type Strain ◽  
Uropathogenic Escherichia Coli ◽  
Wild Type ◽  
Upec Strain ◽  
Host Immune Responses ◽  
Strain Cft073 ◽  
Tract Infections

ABSTRACT Uropathogenic Escherichia coli (UPEC) causes most uncomplicated urinary tract infections (UTIs) in humans. Flagellum-mediated motility and chemotaxis have been suggested to contribute to virulence by enabling UPEC to escape host immune responses and disperse to new sites within the urinary tract. To evaluate their contribution to virulence, six separate flagellar mutations were constructed in UPEC strain CFT073. The mutants constructed were shown to have four different flagellar phenotypes: fliA and fliC mutants do not produce flagella; the flgM mutant has similar levels of extracellular flagellin as the wild type but exhibits less motility than the wild type; the motAB mutant is nonmotile; and the cheW and cheY mutants are motile but nonchemotactic. Virulence was assessed by transurethral independent challenges and cochallenges of CBA mice with the wild type and each mutant. CFU/ml of urine or CFU/g bladder or kidney was determined 3 days postinoculation for the independent challenges and at 6, 16, 48, 60, and 72 h postinoculation for the cochallenges. While these mutants colonized the urinary tract during independent challenge, each of the mutants was outcompeted by the wild-type strain to various degrees at specific time points during cochallenge. Altogether, these results suggest that flagella and flagellum-mediated motility/chemotaxis may not be absolutely required for virulence but that these traits contribute to the fitness of UPEC and therefore significantly enhance the pathogenesis of UTIs caused by UPEC.

Loss of an Intimin-Like Protein Encoded on a Uropathogenic E. coli Pathogenicity Island Reduces Inflammation and Affects Interactions with the Urothelium

2021 ◽  
Author(s):  
Allyson E. Shea ◽  
Jolie A. Stocki ◽  
Stephanie D. Himpsl ◽  
Sara N. Smith ◽  
Harry L. T. Mobley
Keyword(s):  
Ex Vivo ◽  
Cell Adherence ◽  
Upec Strain ◽  
E Coli ◽  
Bladder Cell ◽  
Bladder Cells ◽  
Strain Cft073 ◽  
The Individual ◽  
Tract Infections

Uropathogenic Escherichia coli (UPEC) causes the majority of uncomplicated urinary tract infections (UTI), which affect nearly half of women worldwide. Many UPEC strains encode an annotated intimin-like adhesin ( ila ) locus in their genome related to a well-characterized virulence factor in diarrheagenic E. coli pathotypes. Its role in UPEC uropathogenesis, however, remains unknown. In prototype UPEC strain CFT073, there is an ila locus that encodes three predicted intimin-like genes sinH , sinI , and ratA . We used in silico approaches to determine the phylogeny and genomic distribution of this locus among uropathogens. We found that the currently annotated intimin-encoding proteins in CFT073 are more closely related to invasin proteins found in Salmonella . Deletion of the individual sinH , sinI , and ratA genes did not result in measurable effects on growth, biofilm formation, or motility in vitro . On average, sinH was more highly expressed in clinical strains during active human UTI than in human urine ex vivo . Unexpectedly, we found that strains lacking this ila locus had increased adherence to bladder cells in vitro , coupled with a decrease in bladder cell invasion and death. The sinH mutant displayed a significant fitness defect in the murine model of ascending UTI including reduced inflammation in the bladder. These data confirmed an inhibitory role in bladder cell adherence to facilitate invasion and inflammation; therefore, the ila locus should be termed invasin-like, rather than intimin-like. Collectively, our data suggest that loss of this locus mediates measurable interactions with bladder cells in vitro and contributes to fitness during UTI.

scholarly journals Inhibition of Escherichia coli CFT073fliCExpression and Motility by Cranberry Materials

2011 ◽  
Vol 77 (19) ◽  
pp. 6852-6857 ◽  
Author(s):  
Gabriela Hidalgo ◽  
Michelle Chan ◽  
Nathalie Tufenkji
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Upper Urinary Tract ◽  
Upec Strain ◽  
Microscopy Imaging ◽  
Content Type ◽  
Reverse Transcription Pcr ◽  
Strain Cft073 ◽  
Tract Infections

ABSTRACTIn humans, uropathogenicEscherichia coli(UPEC) is the most common etiological agent of uncomplicated urinary tract infections (UTIs). Cranberry extracts have been linked to the prevention of UTIs for over a century; however, a mechanistic understanding of the way in which cranberry derivatives prevent bacterial infection is still lacking. In this study, we used afliC-luxreporter as well as quantitative reverse transcription-PCR to demonstrate that when UPEC strain CFT073 was grown or exposed to dehydrated, crushed cranberries or to purified cranberry-derived proanthocyanidins (cPACs), expression of the flagellin gene (fliC) was inhibited. In agreement with these results, transmission electron microscopy imaging of bacteria grown in the presence of cranberry materials revealed fewer flagella than those in bacteria grown under control conditions. Furthermore, we showed that swimming and swarming motilities were hindered when bacteria were grown in the presence of the cranberry compounds. Because flagellum-mediated motility has been suggested to enable UPEC to disseminate to the upper urinary tract, we propose that inhibition of flagellum-mediated motility might be a key mechanism by which cPACs prevent UTIs. This is the first report to show that cranberry compounds inhibit UPEC motility via downregulation of thefliCgene. Further studies are required to establish whether these inhibitors play a rolein vivo.

scholarly journals Pectic Oligosaccharides from Cranberry Prevent Quiescence and Persistence in the Uropathogenic Escherichia coli CFT073

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Jiadong Sun ◽  
Robert W. Deering ◽  
Zhiyuan Peng ◽  
Laila Najia ◽  
Christina Khoo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Chemical Compounds ◽  
Uropathogenic Escherichia Coli ◽  
Cranberry Juice ◽  
Persister Cells ◽  
E Coli ◽  
Pectic Oligosaccharides ◽  
Bladder Cells ◽  
Strain Cft073 ◽  
Tract Infections

AbstractUrinary tract infections (UTIs) caused by Escherichia coli create a large burden on healthcare and frequently lead to recurrent infections. Part of the success of E. coli as an uropathogenic bacterium can be attributed to its ability to form quiescent intracellular reservoirs in bladder cells and its persistence after antibiotic treatment. Cranberry juice and related products have been used for the prevention of UTIs with varying degrees of success. In this study, a group of cranberry pectic oligosaccharides (cPOS) were found to both inhibit quiescence and reduce the population of persister cells formed by the uropathogenic strain, CFT073. This is the first report detailing constituents of cranberry with the ability to modulate these important physiological aspects of uropathogenic E. coli. Further studies investigating cranberry should be keen to include oligosaccharides as part of the ‘active’ cocktail of chemical compounds.

scholarly journals Flagella, Type I Fimbriae and Curli of Uropathogenic Escherichia coli Promote the Release of Proinflammatory Cytokines in a Coculture System

2021 ◽  
Vol 9 (11) ◽  
pp. 2233
Author(s):  
Rubí Vega-Hernández ◽  
Sara A. Ochoa ◽  
Ricardo Valle-Rios ◽  
Gustavo A. Jaimes-Ortega ◽  
José Arellano-Galindo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Proinflammatory Cytokines ◽  
Public Health Problem ◽  
Uropathogenic Escherichia Coli ◽  
Type I ◽  
Upec Strain ◽  
Coculture System ◽  
Type I Fimbriae ◽  
High Concentrations ◽  
Tract Infections

Background. Urinary tract infections (UTIs) are a public health problem in Mexico, and uropathogenic Escherichia coli (UPEC) is one of the main etiological agents. Flagella, type I fimbriae, and curli promote the ability of these bacteria to successfully colonize its host. Aim. This study aimed to determine whether flagella-, type I fimbriae-, and curli-expressing UPEC induces the release of proinflammatory cytokines in an established coculture system. Methods. The fliC, fimH, and csgA genes by UPEC strain were disrupted by allelic replacement. Flagella, type I fimbriae, and curli were visualized by transmission electron microscopy (TEM). HTB-5 (upper chamber) and HMC-1 (lower chamber) cells cocultured in Transwell® plates were infected with these UPEC strains and purified proteins. There was adherence to HTB-5 cells treated with different UPEC strains and they were quantified as colony-forming units (CFU)/mL. Results. High concentrations of IL-6 and IL-8 were induced by the FimH and FliC proteins; however, these cytokines were detected in low concentrations in presence of CsgA. Compared with UPEC CFT073, CFT073ΔfimH, CFT073ΔfimHΔfliC, and CFT073ΔcsgAΔfimH strains significantly reduced the adherence to HTB-5 cells. Conclusion. The FimH and FliC proteins are involved in IL-6 and IL-8 release in a coculture model of HTB-5 and HMC-1 cells.

scholarly journals Human Renal Fibroblasts, but Not Renal Epithelial Cells, Induce IL-1β Release during a Uropathogenic Escherichia coli Infection In Vitro

Cells ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3522
Author(s):  
Ashok Kumar Kumawat ◽  
Geena Varghese Paramel ◽  
Kartheyaene Jayaprakash Demirel ◽  
Isak Demirel
Keyword(s):  
Escherichia Coli ◽  
Epithelial Cells ◽  
Uropathogenic Escherichia Coli ◽  
Upec Strain ◽  
Renal Epithelial Cells ◽  
Caspase 1 ◽  
Escherichia Coli Infection ◽  
Strain Cft073 ◽  
Factor Α

Understanding how uropathogenic Escherichia coli (UPEC) modulates the immune response in the kidney is essential to prevent UPEC from reaching the bloodstream and causing urosepsis. The purpose of this study was to elucidate if renal fibroblasts can release IL-1β during a UPEC infection and to investigate the mechanism behind the IL-1β release. We found that the UPEC strain CFT073 induced an increased IL-1β and LDH release from renal fibroblasts, but not from renal epithelial cells. The UPEC-induced IL-1β release was found to be NLRP3, caspase-1, caspase-4, ERK 1/2, cathepsin B and serine protease dependent in renal fibroblasts. We also found that the UPEC virulence factor α-hemolysin was necessary for IL-1β release. Conditioned medium from caspase-1, caspase-4 and NLRP3-deficient renal fibroblasts mediated an increased reactive oxygen species production from neutrophils, but reduced UPEC phagocytosis. Taken together, our study demonstrates that renal fibroblasts, but not renal epithelial cells, release IL-1β during a UPEC infection. This suggest that renal fibroblasts are vital immunoreactive cells and not only structural cells that produce and regulate the extracellular matrix.

scholarly journals Identification ofIn Vivo-Induced Antigens Including an RTX Family Exoprotein Required for Uropathogenic Escherichia coli Virulence

2011 ◽  
Vol 79 (6) ◽  
pp. 2335-2344 ◽  
Author(s):  
Patrick D. Vigil ◽  
Christopher J. Alteri ◽  
Harry L. T. Mobley
Keyword(s):  
Escherichia Coli ◽  
Iron Acquisition ◽  
Expression Library ◽  
Upec Strain ◽  
Infected Female ◽  
Content Type ◽  
Strain Cft073 ◽  
Tract Infections

ABSTRACTUncomplicated urinary tract infections (UTI) are caused most commonly by uropathogenicEscherichia coli(UPEC). Whole-genome screening approaches, including transcriptomic, proteomic, and signature-tagged mutagenesis, have shown that UPEC highly expresses or requires genes for translational machinery, capsule, lipopolysaccharide, type 1 fimbriae, and iron acquisition systems during UTI. To identify additional genes expressed by UPEC during UTI, an immunoscreening approach termedin vivo-induced antigen technology (IVIAT) was employed to identify antigens produced during experimental infection that are not produced duringin vitroculture. An inducible protein expression library, constructed from genomic DNA isolated from UPEC strain CFT073, was screened using exhaustively adsorbed pooled sera from 20 chronically infected female CBA/J mice. Using this approach, we identified 93 antigens induced by UPECin vivo. A representative subset of these genes was tested by quantitative PCR for expression by CFT073in vivoand during growth in human urine or LB mediumin vitro;proWX,narJI,lolA,lolD,tosA(upxA), c2432,katG,ydhX,kpsS, andyddQwere poorly expressedin vitrobut highly expressedin vivo. Of these,tosA, a gene encoding a predicted repeat-in-toxin family member, was expressed exclusively during UTI. Deletion oftosAin UPEC strain CFT073 resulted in significant attenuation in bladder and kidney infections during ascending UTI. By screening forin vivo-induced antigens, we identified a novel UPEC virulence factor and additional proteins that could be useful as potential vaccine targets.

scholarly journals Distribution of Pathogenicity Islands Among Uropathogenic Escherichia coli Isolates From Patients With Urinary Tract Infections

2020 ◽  
Vol 8 (2) ◽  
pp. 39-43
Author(s):  
Ehsan Asadi ◽  
Mohammad Mohammadzadeh ◽  
Mohammad Niakan
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Etiologic Agent ◽  
Uropathogenic Escherichia Coli ◽  
Pathogenicity Islands ◽  
Virulence Determinants ◽  
E Coli ◽  
Virulence Markers ◽  
Microbiological Methods ◽  
Tract Infections

Background: Uropathogenic Escherichia coli (UPEC) is one of the most common etiologic agent of urinary tract infection (UTI). The ability of Escherichia coli to cause UTI is associated with specific virulence determinants, which are encoded by pathogenicity islands (PAIs). Objectives: This study aimed to investigate the distribution of PAIs among the UPEC isolates collected from patients with UTIs. Materials and Methods: In this study, a total of 100 E. coli isolates were collected from patients with UTIs using standard microbiological methods. Polymerase chain reaction (PCR) was used for the identification of the main PAIs of UPEC according to insertion sites and virulence markers. Results: In total, PAI IV536, PAI III536, PAI I536, PAI, IICFT073, PAI ICFT073, PAI IIJ96, PAI II536, and PAI IJ96 were detected in 23, 22, 17, 17, 13, 11, 11, and 8% of isolates. PAI combinations were identified in 15% of isolates. Conclusion: The results showed that PAIs of UPEC are not strain-specific and some strains can carry the PAIs associated with the prototype strains of UPEC simultaneously.

Identification and Ranking of Clinical Compounds with Activity Against Log-phase Growing Uropathogenic Escherichia coli

2020 ◽  
Vol 17 (2) ◽  
pp. 191-196
Author(s):  
Hongxia Niu ◽  
Rebecca Yee ◽  
Peng Cui ◽  
Shuo Zhang ◽  
Lili Tian ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Uropathogenic Escherichia Coli ◽  
Protein Synthesis Inhibitor ◽  
Beta Lactam ◽  
Synthesis Inhibitor ◽  
Upec Strain ◽  
Antibiotic Resistant ◽  
Tetracycline Antibiotics ◽  
Cephalosporin Antibiotics ◽  
Tract Infections

Background: Uropathogenic Escherichia coli (UPEC) is a major cause of Urinary Tract Infections (UTIs). Due to increasing antibiotic-resistance among UPEC bacteria, new treatment options for UTIs are urgently needed. Objective: To identify new agents targeting growing bacteria that may be used for the treatment of antibiotic-resistant UTIs. Methods: We screened a clinical compound library consisting of 1,524 compounds using a high throughput 96-well plate assay and ranked the activities of the selected agents according to their MICs against the UPEC strain UTI89. Results: We identified 33 antibiotics which were active against log-phase clinical UPEC strain UTI89. Among the selected antibiotics, there were 12 fluoroquinolone antibiotics (tosufloxacin, levofloxacin, sparfloxacin, clinafloxacin, pazufloxacin, gatifloxacin, enrofloxacin, lomefloxacin, norfloxacin, fleroxacin, flumequine, ciprofloxacin), 15 beta-lactam or cephalosporin antibiotics (cefmenoxime, cefotaxime, ceftizoxime, cefotiam, cefdinir, cefoperazone, cefpiramide, cefamandole, cefixime, ceftibuten, cefmetazole, cephalosporin C, aztreonam, piperacillintazobactam, mezlocillin), 3 tetracycline antibiotics (meclocycline, doxycycline, tetracycline), 2 membrane-acting agents (colistin and clofoctol), and 1 protein synthesis inhibitor (amikacin). Among them, the top 7 hits were colistin, tosufloxacin, levofloxacin, sparfloxacin, clinafloxacin, cefmenoxime and pazufloxacin, where clinafloxacin and pazufloxacin were the newly identified agents active against UPEC strain UTI89. We validated the key results obtained with UTI89 on two other UTI strains CFT073 and KTE181 and found that they all had comparable MICs for fluoroquinolones while CFT073 and KTE181 were more susceptible to cephalosporin antibiotics and tetracycline antibiotics but were less susceptible to colistin than UTI89. Conclusion: Our findings provide possible effective drug candidates for the more effective treatment of antibiotic-resistant UTIs.

scholarly journals The RyfA small RNA regulates oxidative and osmotic stress responses and virulence in uropathogenic Escherichia coli

2021 ◽  
Vol 17 (5) ◽  
pp. e1009617
Author(s):  
Hicham Bessaiah ◽  
Pravil Pokharel ◽  
Hamza Loucif ◽  
Merve Kulbay ◽  
Charles Sasseville ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Small Rna ◽  
Stress Responses ◽  
Surface Proteins ◽  
Upec Strain ◽  
Expression Of Genes ◽  
Bacterial Infectious Disease ◽  
Strain Cft073 ◽  
Tract Infections

Urinary tract infections (UTIs) are a common bacterial infectious disease in humans, and strains of uropathogenic Escherichia coli (UPEC) are the most frequent cause of UTIs. During infection, UPEC must cope with a variety of stressful conditions in the urinary tract. Here, we demonstrate that the small RNA (sRNA) RyfA of UPEC strains is required for resistance to oxidative and osmotic stresses. Transcriptomic analysis of the ryfA mutant showed changes in expression of genes associated with general stress responses, metabolism, biofilm formation and genes coding for cell surface proteins. Inactivation of ryfA in UPEC strain CFT073 decreased urinary tract colonization in mice and the ryfA mutant also had reduced production of type 1 and P fimbriae (pili), adhesins which are known to be important for UTI. Furthermore, loss of ryfA also reduced UPEC survival in human macrophages. Thus, ryfA plays a key regulatory role in UPEC adaptation to stress, which contributes to UTI and survival in macrophages.

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