Objectives
To examine the in vitro selection of aztreonam/avibactam resistance among MBL-producing Klebsiella pneumoniae and to understand the mechanism of increased resistance.
Methods
The MICs of aztreonam were determined with and without avibactam (4 mg/L) using a broth microdilution method. Single-step and multi-step mutant selection was conducted on five MBL-producing K. pneumoniae strains, including two dual carbapenemase producers. Genomic sequencing and gene cloning were performed to investigate the mechanism of increased resistance.
Results
We examined the MICs for 68 MBL-producing K. pneumoniae isolates, including 13 dual carbapenemase producers. Compared with aztreonam alone, the addition of avibactam (4 mg/L) reduced the MICs for all isolates by >128-fold, with MIC50 and MIC90 values of 0.25 and 1 mg/L, respectively. One NDM-1-, OXA-48-, CTX-M-15- and CMY-16-positive ST101 K. pneumoniae strain was selected to be resistant to aztreonam/avibactam, with a >16-fold increase in MIC (>128 mg/L). WGS revealed that the resistant mutants lost the blaNDM-1 gene, but acquired amino acid substitutions in CMY-16 (Tyr150Ser and Asn346His). Construction of blaCMY-16 mutants confirmed that the substitutions (Tyr150Ser and Asn346His) were primarily responsible for the decreased susceptibility to aztreonam/avibactam. In addition, transfer of blaCMY-16 mutant (Tyr150Ser and Asn346His) plasmid constructs into certain clinical carbapenemase-producing isolates demonstrated >64-fold increased MICs of aztreonam/avibactam and aztreonam/avibactam/ceftazidime.
Conclusions
Aztreonam in combination with avibactam showed potent in vitro activity against MBL-producing K. pneumoniae. However, our study suggested the likelihood of aztreonam/avibactam resistance among MBL- and AmpC-co-producing strains and clinical practice should beware of the possibility of the emerging resistance.
Ceftazidime-avibactam is a potent antibiotic combination against Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacteriaceae. Here, we describe a unique ceftazidime-avibactam-resistant and carbapenem-susceptible K. pneumoniae strain harboring a novel blaKPC-14 variant. This strain was isolated from a New York City patient in 2003, which predates the introduction of avibactam. Despite resistance to ceftazidime-avibactam, the strain was susceptible to imipenem-relebactam and meropenem-vaborbactam. Comprehensive genomic sequencing revealed that blaKPC-14 is harbored on an ST6 IncN plasmid associated with the early spread of blaKPC.
IMPORTANCE KPC is currently the most common carbapenemase identified in the United States. More than 40 KPC variants have been described, of which KPC-2 and KPC-3 are the most frequent clinical variants. However, our understanding of the genetic structures and β-lactam resistance profiles of other novel KPC variants remains incomplete. Here, we report a novel blaKPC variant (blaKPC-14) and the complete genome sequence of blaKPC-14-harboring K. pneumoniae strain BK13048, which is susceptible to carbapenems but resistant to ceftazidime-avibactam. To the best of our knowledge, this is one of the earliest KPC-producing K. pneumoniae strains exhibiting resistance to ceftazidime-avibactam.
To assess the antimicrobial activities of ceftazidime/avibactam (CAZ/AVI) and aztreonam/avibactam (ATM/AVI) against carbapenem-resistant Enterobacteriaceae (CRE) isolates collected from three secondary hospitals in Southwest China between 2018 and 2019. Materials and Methods: A total of 120 unique CRE clinical isolates were collected and carbapenemase genes were detected using PCR. Antimicrobial susceptibility was determined using standard broth microdilution method and the results were interpreted according to CLSI breakpoints. Results: The 120 carbapenem-resistant strains included 92 Klebsiella pneumoniae, 10 Escherichia coli, 10 Enterobacter cloacae, five Klebsiella aerogenes, and three Klebsiella oxytoca isolates. Seventy-four percent of these 120 CRE isolates were collected from patients located in non-ICUs; 65.0% of these CRE isolates were collected from male patients; and 34.2% of these isolates were isolated from respiratory tracts. Four different carbapenemase genes were identified among 103 carbapenemase-producing Enterobacteriaceae (CPE) isolates, including bla KPC-2 (n=77), bla NDM-1 (n=16), bla NDM-5 (n=12) and bla IMP-4 (n=2). Overall, 21.7%, 37.5%, 40.8%, 75.0%, and 100% of the CRE strains were susceptible to levofloxacin, trimethoprim/sulfamethoxazole, amikacin, CAZ/ AVI, and ATM/AVI, respectively. In addition, antimicrobial susceptibility testing showed that 96.7% isolates (n=116) were resistant to aztreonam, and the addition of avibactam (4 mg/L) significantly reduced the MICs of those aztreonam-resistant isolates by more than 128-fold (range: ≤0.125-4 mg/L), and 90.0% (n=108) of total 120 isolates were inhibited at ATM/AVI concentration ≤1 mg/L. Conclusion: Our study revealed significant antimicrobial resistance among the CRE isolates against some commonly used antibiotics in three secondary Chinese hospitals. ATM/AVI exhibited potent activity against CRE isolates, including double carbapenemase-producing isolates, whereas CAZ/AVI was active against all KPC producers.
Liver cancer, the second most commonly diagnosed cancer, is associated with high mortality rates.
E2F4
is a member of the E2F transcription factor family. There are limited studies on the role of
E2F4
in hepatocellular carcinoma (HCC). In this study, the expression of
E2F4
in HCC tissue samples and cell lines was analyzed using quantitative real-time polymerase chain reaction.
E2F4
expression positively correlated with tumor size in patients with HCC. Additionally,
E2F4
expression was greater in HCC cells than in normal LO2 cells. Furthermore, overexpression of
E2F4
significantly enhanced the proliferation, migration, and invasion of HCC cells. The results of a luciferase assay revealed that
E2F4
upregulated the expression of
CDCA3
by binding to its promoter region (1863'-ACGCGCGAGAATG-1875') and consequently promoted proliferation and cell cycle progression of HCC cells. Taken together, these results demonstrated that E2F4 might play a vital role in HCC progression and could serve as a potential biomarker for the diagnosis and as a therapeutic target of HCC.
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