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At least 1,207 records · Page 67Linked to original sources
PubMed1969

Structural studies on colanic acid, the common exopolysaccharide found in the enterobacteriaceae, by partial acid hydrolysis. Oligosaccharides from colanic acid.

The exopolysaccharide slime colanic acid has been isolated from representative strains of Escherichia coli, Salmonella typhimurium and Aerobacter cloacae. Analysis showed that each polymer contained glucose, galactose, fucose and glucuronic acid, together with acetate and pyruvate. The molar proportions of these components were 1:1.8:1.9:1:1:1 approximately. On the basis of periodate oxidation of the natural and deacetylated polysaccharide, glucose is proposed as the site of the acetyl groups. The pyruvate is attached to galactose. Three neutral oligosaccharides and ten electrophoretically mobile oligosaccharides were isolated and partially characterized. Four of the fragments were esters of pyruvic acid. Most oligosaccharides were isolated from all three polysaccharide preparations. Three further oligosaccharides were isolated from carboxyl-reduced colanic acid and sodium borotritide was used to label the glucose derived from glucuronic acid in these fragments. One trisaccharide was obtained from periodate-oxidized polysaccharide. On the basis of these oligosaccharides a repeating hexasaccharide unit of the following structure is proposed: [Formula: see text] The significance of this structure in colanic acid biosynthesis is discussed.

Acetates↗
PubMed1993

Proton-linked L-rhamnose transport, and its comparison with L-fucose transport in Enterobacteriaceae.

1. An alkaline pH change occurred when L-rhamnose, L-mannose or L-lyxose was added to L-rhamnose-grown energy-depleted suspensions of strains of Escherichia coli. This is diagnostic of sugar-H+ symport activity. 2. L-Rhamnose, L-mannose and L-lyxose were inducers of the sugar-H+ symport and of L-[14C]rhamnose transport activity. L-Rhamnose also induced the biochemically and genetically distinct L-fucose-H+ symport activity in strains competent for L-rhamnose metabolism. 3. Steady-state kinetic measurements showed that L-mannose and L-lyxose were competitive inhibitors (alternative substrates) for the L-rhamnose transport system, and that L-galactose and D-arabinose were competitive inhibitors (alternative substrates) for the L-fucose transport system. Additional measurements with other sugars of related structure defined the different substrate specificities of the two transport systems. 4. The relative rates of H+ symport and of sugar metabolism, and the relative values of their kinetic parameters, suggested that the physiological role of the transport activity was primarily for utilization of L-rhamnose, not for L-mannose or L-lyxose. 5. L-Rhamnose transport into subcellular vesicles of E. coli was dependent on respiration, was optimal at pH 7, and was inhibited by protonophores and ionophores. It was insensitive to N-ethylmaleimide or cytochalasin B. 6. L-Rhamnose, L-mannose and L-lyxose each elicited an alkaline pH change when added to energy-depleted suspensions of L-rhamnose-grown Salmonella typhimurium LT2, Klebsiella pneumoniae, Klebsiella aerogenes, Erwinia carotovora carotovora and Erwinia carotovora atroseptica. The relative rates of subsequent acidification varied, depending on both the organism and the sugar. L-Fucose promoted an alkaline pH change in all the L-rhamnose-induced organisms except the Erwinia species. No L-rhamnose-H+ symport occurred in any organism grown on L-fucose. 7. All these results showed that L-rhamnose transport into the micro-organisms occurred by a system different from that for L-fucose transport. Both systems are energized by the trans-membrane electrochemical gradient of protons. 8. Neither steady-state kinetic measurements nor binding-protein assays revealed the existence of a second L-rhamnose transport system in E. coli.

Binding, Competitive↗
PubMed1999

Identification of UreR binding sites in the Enterobacteriaceae plasmid-encoded and Proteus mirabilis urease gene operons.

The closely related Proteus mirabilis and Enterobacterlaceae plasmid-encoded urease genes are positively regulated by the AraC-like transcriptional activator UreR. In the presence of the effector molecule urea, UreR promotes transcription of ureD, the initial gene in the urease operon, and increases transcription of the divergently transcribed ureR. Here, we identify UreR-specific binding sites in the ureRp-ureDp intergenic regions. Recombinant UreR (rUreR) was expressed and purified, and gel shift and DNase I protection assays were performed with this protein. These analyses indicated that there are two distinct rUreR binding sites in both the plasmid-encoded and P. mirabilis ureRp-ureDp intergenic regions. A consensus binding site of TA/GT/CA/TT/GC/TTA/TT/AATTG was predicted from the DNase I protection assays. Although rUreR bound to the specific DNA binding site in both the presence and the absence of urea, the dissociation rate constant k-1 of the rUreR-DNA complex interaction was measurably different when urea was present. In the absence of urea, the dissociation of the protein-DNA complexes, for both ureRp and ureDp, was complete at the earliest time point, and it was not possible to determine a rate. In the presence of urea, dissociation was measurable with a k-1 for the rUreR-ureRp interaction of 1.2 +/- 0.2 x 10(-2) s-1 and a k-1 for the rUreR-ureDp interaction of 2.6 +/- 0.1 x 10(-3) s-1. This corresponds to a half-life of the ureRp-rUreR interaction of 58 s, and a half-life of the ureDp-rUreR interaction of 4 min 26 s. A model describing a potential role for urea in the activation of these promoters is proposed.

Bacterial Proteins↗
PubMed2001

Multiplex PCR for detection of Enterobacteriaceae in blood.

BACKGROUND: Rapid and sensitive methods are needed to detect the small numbers of bacteria that may sometimes contaminate units of blood during collection. A multiplex 5'-nuclease TaqMan PCR assay (PE Applied Biosystems) was used to detect several bacterial species that may contaminate blood. STUDY DESIGN AND METHODS: Oligonucleotide primers were made for regions of the 16S rRNA gene conserved in four different bacterial species: Yersinia enterocolitica and Serratia, Klebsiella, and Enterobacter species. Two probes were designed: SL-1 detected Serratia, Klebsiella, and Enterobacter species, and YE-3 detected Y. enterocolitica. RESULTS: When TaqMan PCR was performed with chromosomal DNA isolated from pure cultures of Serratia liquefaciens, Klebsiella oxytoca, Klebsiella pneumoniae, Enterobacter cloacae, and Enterobacter agglomerans, the limit of detection with probe SL-1 was 1 to 2 CFUs. For S. marcescens, the sensitivity was 8 CFUs. The limit of detection for Y. enterocolitica with probe YE-3 was 2 CFUs. When total chromosomal DNA was extracted from whole-blood samples spiked with different numbers of Y. enterocolitica, S. liquefaciens, E. cloacae, or K. pneumoniae bacteria, the TaqMan PCR detected 12 to 16 organisms in 1 mL of blood. CONCLUSION: The 5'-nuclease TaqMan PCR assay takes only 3 hours to perform and has the potential to detect very small numbers of bacteria.

Blood↗
PubMed2002

Characterisation of local isolates of Enterobacteriaceae from Turkey.

20 local isolates of enterics belonging to the genera Salmonella, Enterobacter, Proteus, Citrobacter from human, chicken and/or egg were characterised for their antibiotic resistance patterns, plasmid profiles, phage types, outer membrane proteins, and lipopolysaccharide patterns. Relatedness of these characteristics for epidemiological analysis was assessed. 18 (90%) strains were resistant to at least one antibiotic and those (multi-drug resistant ones) resisting to two or more antibiotics constituted 50% of all isolates. A common 54 kb plasmid was harboured by 55% of the isolates. 14 isolates showed smooth type lipopolysaccharide. 60% of the 20 isolates contained outer membrane proteins in a molecular weight range of 34.6 to 30.6 kDa. The data reveal the lack of correlation between the characteristics investigated.

Animals↗
PubMed1998

The effects of the morphological response of Enterobacteriaceae to cephalosporins on PAE and CERT.

PAE and CERT values of cefaclor, loracarbef and cefuroxime (0.1-100 x MIC) were established for 8 E. coli, 3 K. pneumoniae and 2 P. mirabilis isolates. Cell enumeration was by impedance (IMP) monitoring in combination with either bioluminescence (BIOL) or viable counting (VC). Morphology was determined by interference contrast microscopy. After 2 h exposure to cefaclor, loracarbef and cefuroxime; concentration-dependent differences in counts were seen by BIOL and VC, varying from a mean value of 0.07 x log10 after exposing the P. mirabilis isolates to 0.1 x MIC cefaclor to a mean value of 2.24 x log10 after exposing the E. coli strains to 100 x MIC cefuroxime. Higher concentrations gave rise to fragile morphological forms including spheroplasts and lower concentrations to less fragile forms such as long bacilli. The longest PAE and CERT values were obtained after exposing the E. coli strains to 100 x MIC cefaclor with mean values of 4.07 and 4.87 h, respectively. Corresponding values were PAE and CERT values of 2.17 and 2.60 h for 100 x cefuroxime and 3.45 and 2.91 h for 100 x MIC loracarbef. By the Student's t-test, PAE values determined by IMP/BIOL and IMP/VC were found to be significantly different, whereas CERT values were found not to be significantly different. PAE and CERT are concentration dependent and vary with specific antibiotic/organism combinations.

Cefaclor↗
PubMed2006

Limiting the emergence of extended-spectrum Beta-lactamase-producing enterobacteriaceae: influence of patient population characteristics on the response to antimicrobial formulary interventions.

BACKGROUND: Effective methods to control the emergence of extended-spectrum beta -lactamase-producing Escherichia coli and Klebsiella species (ESBL-EK) remain unclear. Variations in the patient populations at different hospitals may influence the effect of antimicrobial formulary interventions. METHODS: To examine variations across hospitals in the response to antimicrobial interventions (ie, restriction of ceftazidime and ceftriaxone) designed to curb the spread of ESBL-EK, we conducted a 5-year quasi-experimental study. This study was conducted at 2 hospitals within the same health system: Hospital A is a 625-bed academic medical center, and Hospital B is a 344-bed urban community hospital. All adult patients with a healthcare-acquired clinical culture of ESBL-EK from July 1, 1997 through December 31, 2002 were included. RESULTS: After the interventions, the use of ceftriaxone decreased by 86% at Hospital A and by 95% at Hospital B, whereas the use of ceftazidime decreased by 95% at Hospital A and by 97% at Hospital B. The prevalence of ESBL-EK at Hospital A decreased by 45% (P < .001), compared with a 22% decrease at Hospital B (P = .36). The following variables were significantly more common among ESBL-EK-infected patients at Hospital B: residence in a long-term care facility (adjusted odds ratio, 3.77 [95% confidence interval, 1.70-8.37]), advanced age (adjusted odds ratio, 1.04 [95% confidence interval, 1.01-1.06]), and presence of a decubitus ulcer (adjusted odds ratio, 4.13 [95% confidence interval, 1.97-8.65]). CONCLUSIONS: The effect of antimicrobial formulary interventions intended to curb emergence of ESBL-EK may differ substantially across institutions, perhaps as a result of differences in patient populations. Variability in the epidemiological profiles of ESBL-EK isolates at different hospitals must be considered when designing interventions to respond to these pathogens.

Adolescent↗
PubMed2006

Survival of cephalosporin-resistant enterobacteriaceae on fingers.

Five strains of Enterobacter species (n=4) and Pantoea species (n=1) resistant to third-generation cephalosporins and isolated from clusters of 3-25 premature infants and small children and 5 strains (4 Enterobacter strains and 1 Pantoea strain) with the same resistance pattern that were isolated from 1 premature infant or small child each were inoculated on the fingertips of 10 volunteer study participants to test whether survival on fingertips is correlated with horizontal transmission. Although there was no significant difference in survival between the groups of transmitted and sporadic strains, there were significant differences in bacterial survival between the participants.

Cephalosporins↗