No abstract available.
Colon* ; Uropathogenic Escherichia coli*

No abstract available.
Korea* ; Uropathogenic Escherichia coli*

The species Escherichia coli is a large and heterogeneous group of organisms which is the most common etiologic agent of urinary tract infections. There are some evidences that certain properties are found more commonly in strains from the patients with urinary tract infections than in those from normal feces and in enteropathogenic strains. Twenty eight strains of E. coli from acute pyelonephritis patients and twenty eight strains from a cute cystitis patients were examined for the abilities of adhesion to normal uroepitheliaI cells, hemagglutination, hemolysin production and resistance to the bactericidal effect of normal rabbit serum. For comparison, 20 strains from normal feces and 20 enteropathogenic strains were tested for these properties. The results were as follows; l. The strains from the patients with urinary tract infections adhered in significantly larger numbers to the epithelial cells than those from normal feces and enteropathogenic strains (p<0.0l). 2. The strains from the patients with urinary tract infections induced hemagglutination of human erythrocytes no more frequently than those from normal feces. 3. The strains from acute pyelonephritis patients more commonly produced hemolysin than those from normal feces and enteropathogenic strains. 4. The resistance to the bactericidal effect of normal rabbit serum was higher in the strains from the patients with urinary tract infections than in those from normal feces and enteropathogenic stains. 5. There was no significant mutual relationships among the properties of uroepithelial adhesion, hemaglutination, hemolysin production and serum resistance. From the results the abilities of adhesion to human uroepitheliaI cells, hemolysin production and resistance to the bactericidal effect of normal rabbit serum are supposed to be common properties of uropathogenic Escherichia coli and essential factors in the pathogenesis of urinary tract infections.
Coloring Agents ; Cystitis ; Epithelial Cells ; Erythrocytes ; Escherichia coli ; Feces ; Hemagglutination* ; Humans ; Pyelonephritis ; Urinary Tract Infections ; Uropathogenic Escherichia coli*

To investigate the inflammatory cytokine production of human epithelial cell lines stimulated with uropathogenic E. coli strains showing 3 different adherence patterns, differential expression of inflammatory cytokine (IL-1a, IL-lB, IL-8, TNFa, and TGFB) mRNA were detected by RT-PCR. IL-1a, IL-1B, IL-8, and TGFB mRNAs constitutively expressed in epithelial cell lines, but not TNFa. The expression of IL-1a and IL-1B mRNA was increased in J-82 cells stimulated with E. coli strains showing DA, LA, or AggA pattern. The expression of IL-8 mRNA was increased, whereas TGFj3 mRNA was decreased in J-82 cells stimulated with E. coli strain showing AggA.pattern. Treatment with crude bacterial adhesins (CBA) isolated from E. coli strains showing DA or LA pattern increased IL-la, IL-lB, IL-S, and TGFj3 mRNA expressions in J-82 cells and HeLa cells. IL-la, IL-lB, and TGFB mRNA expressions were decreased in epitheUal cells stimulated with CBA from E. coli strain showing AggA pattern, whereas IL-8 mRNA expression was significantly increased. The expressions of cytokine mRNAs showed little differences between epithelial ceRs used, but great differences between CBA from DA or LA and AggA strain. LPS stimulation was little changed cytokine mRNA expressions in epithelial cells. This study suggests that cytokine gene expression of epithelial cells by the bacterial stimulation mainly depends on the bacterial adhesins recognized by the respective receptors of epithelial cells.
Adhesins, Bacterial ; Epithelial Cells* ; Gene Expression ; HeLa Cells ; Humans ; Interleukin-8 ; RNA, Messenger ; Uropathogenic Escherichia coli*

Selected number of urinary E. coli isolates from patients were subjected to the tests to characterize in vitro properties, compared with the strains of normal flora in intestinal tract or other sources. Further studies were carried out for determination of the mechanism of pathogenesis by using rabbit as an experimental animal model. Both strains of 87E1 and 87E51 which had one type of pili, were eliminated rapidly 4 or 8 days after inoculation into renal pelvis of rabbit. On the contrary, 88E18 strain which had both types of pili successfully colonized in urinary tract and the rabbit died due to urosepsis after the 8th day of experiment. The result of bacterial count colonized in the tissue by colony forming unit (CFU)per gram were similar to those in the urine.
Bacterial Load ; Colon ; Humans ; Kidney Pelvis ; Models, Animal ; Stem Cells ; Urinary Tract ; Uropathogenic Escherichia coli* ; Virulence*

Adhesins, Escherichia coli ; genetics ; Bacterial Typing Techniques ; Escherichia coli Proteins ; genetics ; Fimbriae Proteins ; genetics ; Genes, Bacterial ; Genotype ; Uropathogenic Escherichia coli ; classification ; genetics

BACKGROUND: We identified virulence genes in uropathogenic E. coli isolates and studied the association between virulence gene and clinical characteristics in order to predict the severity and recurrency. METHODS: 39 Escherichia coli strains from patients with urinary tract infection were clinically and genotypically characterized. The strains were examined genotypically by using the multiplex polymerase chain reaction for presence of 5 urovirulence genes : pyelonephritis-associated pili(pap), S. fimbriae(sfa), afimbrial adhesin(afa), cytotoxic necrotizing factor (cnf), and alpha-hemolysin(hly). The patient's clinical characteristics were determined retrospectively. RESULTS: 17 pap(+), 4 sfa(+), 7 afa(+), 6 cnf(+), and 8 hly(+) strains were identified. And there were 10 genotypes. Among them, genotype pap(+)sfa(-)afa(-)cnf(-) hly(-) was most dominant(36%). But no urovirulence gene was detected in 12 strains(31%). When the data was analyzed, it was apparent that an association among various urovirulence genes exists. sfa gene was frequently associated with cnf gene(p < 0.001). And afa gene was associated cnf and hly gene(p= 0.026, <0.001). An association between cnf gene and hly gene was observed(p=0.002). Positive rates of virulence genes were not different between male and female. In infancy, pap(-)sfa(-)afa(+)cnf(+)hly(+) genotype was dominant. In 2-15 years old age group, pap(-) sfa(-)afa(-)cnf(-)hly(-) genotype was dominant. And in 16- 40 years old age group, pap(+)sfa(-)afa(-)cnf(-)hly(-) was dominant. So, some virulence genotype might be associated with specific age group. Presence of virulence gene or specific genotype was not different among diseases(acute pyelonephritis, cystitis, asymptomatic bacteriuria). So, virulence genes were not associated with severity of urinary tract infection. Virulence genes were not associated with susceptibility of recurrent infection. In neurogenic bladder patients, there were significantly more sfa(+) strains (p=0.019). And all isolates of neurogenic bladder patients were genotype pap(+)sfa(+)afa(-)cnf(+)hly(-)(p < 0.001). CONCLUSION: In this study, We found which genotype is most dominant in uropathogenic Escherichia coli, and that virulence genes do not suggest severity or recurrency of urinary tract infection. In neurogenic bladder patients, some virulence genes were more prevalent.
Adult ; Cystitis ; Escherichia coli ; Female ; Genotype ; Humans ; Male ; Multiplex Polymerase Chain Reaction ; Polymerase Chain Reaction ; Pyelonephritis ; Retrospective Studies ; Urinary Bladder, Neurogenic ; Urinary Tract Infections ; Uropathogenic Escherichia coli* ; Virulence*

To investigate the effect and the mechanism of ppk1 gene deletion on the drug susceptibility of uropathogenic Escherichia coli producing extended-spectrum beta-lactamases (ESBLs-UPEC). The study was an experimental study. From March to April 2021, a strain of ESBLs-UPEC (genotype was TEM combined with CTX-M-14) named as UE210113, was isolated from urine sample of the patient with urinary tract infection in the Laboratory Department of Guangzhou Eighth People's Hospital, meanwhile its ppk1 gene knock-out strain Δpk1 and complemented strain Δpk1-C were constructed by suicide plasmid homologous recombination technique, which was used to study the effect of ppk1 gene on ESBLs-UPEC drug sensitivity and its mechanism. The drug susceptibility of UE210113, Δpk1, and Δpk1-C were measured by Vitek2 Compact System and broth microdilution method. The quantitative expression of ESBLs, outer membrane protein and multidrug efflux systems encoding genes of UE210113, Δpk1 and Δpk1-C were performed by using qRT-PCR analysis. By using two independent sample Mann-Whitney U test, the drug susceptibility results showed that, compared with UE210113 strain, the sensitivities of Δpk1 to ceftazidime, cefepime, tobramycin, minocycline and cotrimoxazole were enhanced (Z=-2.121,P<0.05;Z=-2.236,P<0.05;Z=-2.236,P<0.05;Z=-2.121,P<0.05), and the drug susceptibility of Δpk1-C restored to the same as which of UE210113 (Z=0,P>0.05). The expression levels of ESBLs-enconding genes bla_TEM and bla_CTX-M-14 in Δpk1 were significantly down-regulated compared with UE210113, but the expression was not restored in Δpk1-C. The expression of outer membrane protein gene omp F in Δpk1 was significantly up-regulated, while the expression of omp A and omp C were down-regulated. The results showed that the expression of multidrug efflux systems encoding genes tol C, mdt A and mdtG were down-regulated in Δpk1 compared with UE210113. The expression of all of the outer membrane protein genes and the multidrug efflux systems genes were restored in Δpk1-C. In conclusion,the lost of ppk1 gene can affect the expression of the outer membrane protein and multidrug efflux systems encoding genes of ESBLs-UPEC, which increase the sensitivity of ESBLs-UPEC to various drugs.
Humans ; beta-Lactamases/metabolism* ; Uropathogenic Escherichia coli/metabolism* ; Urinary Tract Infections ; Plasmids ; Membrane Proteins/genetics* ; Escherichia coli Infections ; Microbial Sensitivity Tests ; Anti-Bacterial Agents/pharmacology*

To investigate the effect and the mechanism of ppk1 gene deletion on the drug susceptibility of uropathogenic Escherichia coli producing extended-spectrum beta-lactamases (ESBLs-UPEC). The study was an experimental study. From March to April 2021, a strain of ESBLs-UPEC (genotype was TEM combined with CTX-M-14) named as UE210113, was isolated from urine sample of the patient with urinary tract infection in the Laboratory Department of Guangzhou Eighth People's Hospital, meanwhile its ppk1 gene knock-out strain Δpk1 and complemented strain Δpk1-C were constructed by suicide plasmid homologous recombination technique, which was used to study the effect of ppk1 gene on ESBLs-UPEC drug sensitivity and its mechanism. The drug susceptibility of UE210113, Δpk1, and Δpk1-C were measured by Vitek2 Compact System and broth microdilution method. The quantitative expression of ESBLs, outer membrane protein and multidrug efflux systems encoding genes of UE210113, Δpk1 and Δpk1-C were performed by using qRT-PCR analysis. By using two independent sample Mann-Whitney U test, the drug susceptibility results showed that, compared with UE210113 strain, the sensitivities of Δpk1 to ceftazidime, cefepime, tobramycin, minocycline and cotrimoxazole were enhanced (Z=-2.121,P<0.05;Z=-2.236,P<0.05;Z=-2.236,P<0.05;Z=-2.121,P<0.05), and the drug susceptibility of Δpk1-C restored to the same as which of UE210113 (Z=0,P>0.05). The expression levels of ESBLs-enconding genes bla_TEM and bla_CTX-M-14 in Δpk1 were significantly down-regulated compared with UE210113, but the expression was not restored in Δpk1-C. The expression of outer membrane protein gene omp F in Δpk1 was significantly up-regulated, while the expression of omp A and omp C were down-regulated. The results showed that the expression of multidrug efflux systems encoding genes tol C, mdt A and mdtG were down-regulated in Δpk1 compared with UE210113. The expression of all of the outer membrane protein genes and the multidrug efflux systems genes were restored in Δpk1-C. In conclusion,the lost of ppk1 gene can affect the expression of the outer membrane protein and multidrug efflux systems encoding genes of ESBLs-UPEC, which increase the sensitivity of ESBLs-UPEC to various drugs.
Humans ; beta-Lactamases/metabolism* ; Uropathogenic Escherichia coli/metabolism* ; Urinary Tract Infections ; Plasmids ; Membrane Proteins/genetics* ; Escherichia coli Infections ; Microbial Sensitivity Tests ; Anti-Bacterial Agents/pharmacology*

The papA gene in uropathogenic Escherichia coli strains was characterized by PCR-restriction fragment length polymorphism (RFLP) and enterobacterial repetitive intergenic consensus (ERIC)- PCR. One hundred four E. coli strains from patients with urinary tract infections and 32 strains from healthy persons were examined. Thirty seven (27.2%) strains (33 from patients, 4 from healthy persons) in the 136 E. coli strains were positive in mannose resistant hemagglutination (MRHA) test. The adherence of MRHA positive strains to HEp-2 cells was greater than those of MRHA negative isolates (p<0.001). PapA-PCR were positive in 25% (26/104) of the strains from patients, and 3.1% (1/32) of the strains from healthy persons. Among 27 papA-positive isolates, subtypes were identified by RFLP as 8 (29.6%) F7z, 3 (11.1%) F9, 4 (14.8%) F12, and 4 (14.8%) F13. Six groups with novel RFLP patterns were detected, also. The subtypes of P-fimbriae was highly similar to each other by ERIC-PCR.
Consensus ; Hemagglutination ; Humans ; Mannose ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Urinary Tract Infections ; Uropathogenic Escherichia coli*