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Find video protocols related to scientific articles indexed in Pubmed.
Predominance of carbapenem resistant Pseudomonas aeruginosa isolates carrying both blaIMP and blaVIM metallo-?-lactamases in a major hospital in Costa Rica.
J. Med. Microbiol.
PUBLISHED: 10-31-2014
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This study aimed to assess the molecular basis of the resistance to carbapenems in clinical isolates of Pseudomonas aeruginosa recovered from a tertiary-level health facility in San José, Costa Rica. To this end, a total of 198 non-duplicated isolates were evaluated for their susceptibility to ?-lactams, aminoglycosides, and fluoroquinolones, the production of metallo-?-lactamases (MBLs), the presence of the blaIMP, blaVIM, and blaGIM-1 MBLs genes, and their occurrence on class 1 integrons. In addition, an ERIC2 PCR fingerprinting method was used to elucidate the distribution of the detected MBL genes within the strain collection. One hundred twenty-five isolates out of 198 (63.1%) were categorized as carbapenem-resistant. The majority (88.8%) of the carbapemen-resistant isolates also showed resistance to ceftazidime, cefepime, aztreonam, ticarcillin/clavulanic acid, amikacin, gentamicin, tobramycin, ciprofloxacin, and gatifloxacin. We found 102 MBL producers (81.6%) among the carbapenem-resistant isolates. Strikingly, both blaIMP and blaVIM genes were simultaneously detected in most (94.1%) of the 102 MBL producers. Five carbapenem-resistant MBL-producers were positive only for blaIMP genes. Almost 70% of the isolates examined harbored the intI1 gene, accompanied by the sul1 and qacE?1 genes in 136 (99%) and 122 (89%) isolates, respectively. The majority (94.4%) of the carbapenem-resistant isolates carried the intI1 gene, in contrast to 26.0% of the carbapenem-susceptible isolates. Ninety-three out of 96 (96.9%) isolates carrying both blaIMP and blaVIM genes also harbored the intI1, sul1, and qacE?1 genes. Gene cassettes from carbapenem-susceptible and MBL-negative carbapenem-resistant isolates encode for aminoglycoside-resistance enzymes (aadA2, aadA4, aadA6) as well as orfD, and qacF genes. RAPD analysis distributed 126 of the isolates in 29 clusters. Eighty of the 90 blaIMP+ blaVIM+ isolates were sorted in 16 different clusters, suggesting that the blaIMP and blaVIM genes detected are probably linked in a genetic structure susceptible of lateral transfer. Carbapenem-resistant MBL-positive isolates were recovered from almost all hospital wards and were over-represented in samples obtained from the surgical emergency and intensive care therapy units. Remarkably, three carbapenem-resistant isolates, exhibiting MBL activity and carrying both blaIMP and blaVIM genes, were recovered from outpatients. Sequence analysis of both bla genes in various isolates revealed that they correspond to the alleles blaIMP-18 and blaVIM-2. To our knowledge, this is the first report of the combination of two metallo-?-lactamases encoded by the blaIMP-18 and blaVIM-2 genes in P. aeruginosa.
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Characterization of Urinary Tract Infection-Associated Shiga Toxin-Producing Escherichia coli.
Infect. Immun.
PUBLISHED: 08-25-2014
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Enterohemorrhagic Escherichia coli (EHEC), a subgroup of Shiga toxin (Stx)-producing E. coli (STEC), is a leading cause of diarrhea and hemolytic-uremic syndrome (HUS) in humans. However, urinary tract infections (UTIs) caused by this microorganism but not associated with diarrhea have occasionally been reported. We geno- and phenotypically characterized three EHEC isolates obtained from the urine of hospitalized patients suffering from UTIs. These isolates carried typical EHEC virulence markers and belonged to HUS-associated E. coli (HUSEC) clones, but they lacked virulence markers typical of uropathogenic E. coli. One isolate exhibited a localized adherence (LA)-like pattern on T24 urinary bladder epithelial cells. Since the glycosphingolipids (GSLs) globotriaosylceramide (Gb3Cer) and globotetraosylceramide (Gb4Cer) are well-known receptors for Stx but also for P fimbriae, a major virulence factor of extraintestinal pathogenic E. coli (ExPEC), the expression of Gb3Cer and Gb4Cer by T24 cells and in murine urinary bladder tissue was examined by thin-layer chromatography and mass spectrometry. We provide data indicating that Stxs released by the EHEC isolates bind to Gb3Cer and Gb4Cer isolated from T24 cells, which were susceptible to Stx. All three EHEC isolates expressed stx genes upon growth in urine. Two strains were able to cause UTI in a murine infection model and could not be outcompeted in urine in vitro by typical uropathogenic E. coli isolates. Our results indicate that despite the lack of ExPEC virulence markers, EHEC variants may exhibit in certain suitable hosts, e.g., in hospital patients, a uropathogenic potential. The contribution of EHEC virulence factors to uropathogenesis remains to be further investigated.
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Characterization of Escherichia coli isolates from hospital inpatients or outpatients with urinary tract infection.
J. Clin. Microbiol.
PUBLISHED: 01-31-2014
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Uropathogenic Escherichia coli (UPEC) is the most common cause of community- and hospital-acquired urinary tract infections (UTIs). Isolates from uncomplicated community-acquired UTIs express a variety of virulence traits that promote the efficient colonization of the urinary tract. In contrast, nosocomial UTIs can be caused by E. coli strains that differ in their virulence traits from the community-acquired UTI isolates. UPEC virulence markers are used to distinguish these facultative extraintestinal pathogens, which belong to the intestinal flora of many healthy individuals, from intestinal pathogenic E. coli (IPEC). IPEC is a diarrheagenic pathogen with a characteristic virulence gene set that is absent in UPEC. Here, we characterized 265 isolates from patients with UTIs during inpatient or outpatient treatment at a hospital regarding their phylogenies and IPEC or UPEC virulence traits. Interestingly, 28 of these isolates (10.6%) carried typical IPEC virulence genes that are characteristic of enteroaggregative E. coli (EAEC), Shiga toxin-producing E. coli (STEC), and atypical enteropathogenic E. coli (aEPEC), although IPEC is not considered a uropathogen. Twenty-three isolates harbored the astA gene coding for the EAEC heat-stable enterotoxin 1 (EAST1), and most of them carried virulence genes that are characteristic of UPEC and/or EAEC. Our results indicate that UPEC isolates from hospital patients differ from archetypal community-acquired isolates from uncomplicated UTIs by their spectrum of virulence traits. They represent a diverse group, including EAEC, as well as other IPEC pathotypes, which in addition contain typical UPEC virulence genes. The combination of typical extraintestinal pathogenic E. coli (ExPEC) and IPEC virulence determinants in some isolates demonstrates the marked genome plasticity of E. coli and calls for a reevaluation of the strict pathotype classification of EAEC.
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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.