In Vivo Pharmacodynamics of ß-Lactams/Nacubactam against Carbapenem-Resistant and/or Carbapenemase-Producing Enterobacter cloacae and Klebsiella pneumoniae in Murine Pneumonia Model.

Carbapenem-resistant Enterobacterales (CRE) and carbapenemase-producing Enterobacterales (CPE) have become global threats. CRE- and CPE- derived infections have been associated with high mortality due to limited treatment options. Nacubactam is a ß-lactamase inhibitor and belongs to the new class of diazabicyclooctane. The agent has an in vitro antimicrobial activity against several classes of ß-lactamase-producing Enterobacterales. This study evaluated antimicrobial activity of combination therapies including ß-lactams (aztreonam, cefepime, and meropenem) and nacubactam against four Enterobacter cloacae and six Klebsiella pneumoniae isolates with murine pneumonia model. Based on changes in bacterial quantity, antimicrobial activities of some regimens were assessed. Combination therapies including ß-lactams (aztreonam, cefepime, and meropenem) with nacubactam showed enhanced antimicrobial activity against CRE E. cloacae (-3.70 to -2.08 Δlog10 CFU/lungs) and K. pneumoniae (-4.24 to 1.47 Δlog10 CFU/lungs) with IMP-1, IMP-6, or KPC genes, compared with aztreonam, cefepime, meropenem, and nacubactam monotherapies. Most combination therapies showed bacteriostatic (-3.0 to 0 Δlog10 CFU/lungs) to bactericidal (<-3.0 Δlog10 CFU/lungs) activities against CRE isolates. This study revealed that combination regimens with ß-lactams (aztreonam, cefepime, and meropenem) and nacubactam are preferable candidates to treat pneumonia due to CRE and CPE.

Pharmacodynamic evaluation of meropenem, cefepime, or aztreonam combined with a novel ß-lactamase inhibitor, nacubactam, against carbapenem-resistant and/or carbapenemase-producing Klebsiella pneumoniae and Escherichia coli using a murine thigh-infection model.

BACKGROUND: Carbapenem-resistant Enterobacterales (CRE) and carbapenemase-producing Enterobacterales (CPE) are difficult to treat and are a serious public health threat. Nacubactam (NAC) is a novel non-ß-lactam diazabicyclooctane ß-lactamase inhibitor with in vitro activity against some Enterobacterales expressing classes of ß-lactamases. METHODS: The antimicrobial efficacy of meropenem (MEM), cefepime (FEP), and aztreonam (ATM), each in combination with NAC, were assessed in vitro and in vivo against Klebsiella pneumoniae and Escherichia coli. Ten isolates, including CRE and/or CPE with ß-lactamase genes, were used in this study. The relationship between phenotype and in vivo efficacy was assessed in a murine neutropenic thigh-infection model. Efficacy was determined by the change in bacterial quantity. RESULTS: The results of the in vitro study showed the minimum inhibitory concentrations of the combination of NAC with either MEM, FEP, or ATM in a 1:1 ratio were 2 to >128-fold lower than those of MEM, FEP, or ATM alone against CRE+ isolates. In addition, combinations of ß-lactams and NAC administered in the murine thigh-infection model showed greater efficacy against CRE+/CPE+, CRE+/CPE-, and CRE-/CPE+ isolates harboring various ß-lactamase genes (IMP-1, IMP-6, KPC, DHA-1, or OXA-48) compared with MEM, FEP, ATM, and NAC alone. CONCLUSION: MEM, FEP, or ATM in combination with NAC showed potent in vivo antimicrobial activity in a murine thigh-infection model caused by K. pneumoniae and E. coli, including CRE and/or CPE isolates. These findings indicate that these combinations of ß-lactams and NAC are potential candidates for the treatment of CRE and/or CPE infections.

Impact of the pneumococcal conjugate vaccine on serotype distribution of adult non-invasive Streptococcus pneumoniae isolates in Tokai region, Japan, 2008-2016.

The introduction of the 7-valent pneumococcal conjugate vaccine (PCV7) in February 2010 markedly reduced the burden of invasive pneumococcal disease and changed serotype distribution in Japan. In November 2013, PCV7 was replaced by the 13-valent pneumococcal conjugate vaccine (PCV13). We investigated the serotype distribution and susceptibility trends of non-invasive Streptococcus pneumoniae isolates collected from adult patients. A total of 504 pneumococcal isolates were collected during 4 periods between 2008 and 2016 (Period 1; between June 2008 and April 2009, Period 2; between September 2010 and March 2011, Period 3; between October 2011 and March 2012, Period 4; between August 2015 and January 2016). The coverage of PCV7 and PCV13 significantly decreased from 38.6% and 60.5% in Period 1 to 6.6% and 31.1% in Period 4. This change was mainly due to a large decrease in the frequency of serotype 19F, 6B, and 14. Serotype 3 was the most frequently isolated, and gradually increased. Additionally, non-PCV13 serotypes 11A, 33F, and 35B significantly increased. Most of the PCV7 serotypes 19F, 23F, 6B, and 14 had mutations of penicillin-binding protein genes and macrolide resistance genes, and these serotypes showed low susceptibilities to cefdinir and clarithromycin. On the other hand, a significant change in susceptibility to various antimicrobial agents was not observed throughout the study period, except for decreased susceptibility to carbapenems. Continuous surveillance studies of pneumococcal serotype changes and drug susceptibility are necessary in future.

Sensitivity surveillance of Pseudomonas aeruginosa isolates for several antibacterial agents in Chubu area (2013-2014).

We investigated the susceptibility to antibacterial agents of 186 clinical isolates of Pseudomonas aeruginosa isolated from medical facilities in Gifu, Aichi, Toyama, and Fukui prefectures from October 2013 to February 2014. MIC50/90 of piperacillin (PIPC), tazobactam/piperacillin (TAZ/PIPC), ceftazidime (CAZ), cefepime (CFPM), imipenem (IPM), meropenem (MEPM), doripenem (DRPM), aztreonam (AZT), ciprofloxacin (CPFX), levofloxacin (LVFX), amikacin (AMK) and colistin (CL) against P aeruginosa was 8/32, 4/32, 2/8, 2/16, 1/32, 0.5/8, 0.25/4, 8/32, 0.25/8, 0.5/16, 4/8 and 1/1pg/mLrespectively. Two strains of multidrug resistant P aeruginosa were isolated (1.1%). They were isolated from the respiratory tract, intra-abdominal, and urinary infection. The susceptible ratio against P aeruginosa derived from intra-abdominal infection for carbapenem was lower than those from respiratory tract and urinary infection. The susceptible ratio against P aeruginosa derived from urinary infection for penicillin, cephem, monobactam, and fluoroquinolone was lower than those from respiratory and intra-abdominal infection. It is meaningful to pay attention to the susceptibility to antibacterial agents in each clinical specimen from infected organ.

Characterization of plasmid-mediated quinolone resistance determinants in Klebsiella pneumoniae and Escherichia coli from Tokai, Japan.

The spread of plasmid-mediated quinolone resistance (PMQR) determinants was evaluated in 150 ceftazidime or cefotaxime-resistant clinical isolates of Klebsiella pneumoniae and Escherichia coli from Tokai, Japan between 2008 and 2011. In this study, qnrB, qnrS, and aac(6')-Ib-cr genes were detected in 12 (50.0%), 4 (16.7%), and 1 (4.2%) of 24 K. pneumoniae isolates, respectively, while qnrA, aac(6')-Ib-cr, and qepA genes were detected in 1 (0.8%), 11 (8.7%), and 2 (1.6%) of 126 E. coli isolates, respectively. qnr genes were mainly found in K. pneumoniae (66.7%) and to a lesser extent in E. coli (0.8%). We determined the genetic environment of the qnrB4 gene in 24.6 kb class 1 integron structure, including aar-2, cmlA, blaOXA-10, aadA1, qacEdelta1, sul1, and blaDHA-1. In a time-kill assay, introduction of the qnrB4 or qnrS1 plasmid to the recipient E. coli strain decreased the bactericidal activities of fluoroquinolones such as ciprofloxacin, levofloxacin, and pazufloxacin.

Impact of the pneumococcal conjugate vaccine on serotype distribution and susceptibility trends of pediatric non-invasive Streptococcus pneumoniae isolates in Tokai, Japan over a 5-year period.

Introduction of the 7-valent pneumococcal conjugate vaccine (PCV7) in February 2010 markedly reduced the burden of invasive pneumococcal disease (IPD) and changed serotype distribution in Japan. We investigated the serotype distribution and susceptibility trends of non-invasive Streptococcus pneumoniae isolates collected from pediatric patients. A total of 564 pneumococcal isolates were collected over a 5-year period between 2008 and 2012. The coverage of PCV7 significantly decreased throughout the study period, from 49.3% in period 1 (between June 2008 and April 2009) to 23.4% in period 4 (between October 2011 and March 2012). This change was mainly due to a large decrease in the frequency of 19F (from 20.6% to 9.9%) and 6B (from 10.3% to 2.7%) and an increase in serotype 3 (from 5.1% to 13.5%) and serogroup 15 (from 4.4% to 9.0%). According to serotype replacement, the susceptible ratios of S. pneumoniae to ß-lactams increased slightly while macrolide resistance remained high. The high frequency of macrolide-resistant pneumococcal isolates may continue because of the high frequency of erm(B) in replace serotypes such as serotype 3 and serogroup 15. The continuous surveillance study is essential following the introduction of a second generation 13-valent pneumococcal conjugate vaccine (PCV13).

Turnip mosaic virus genome-linked protein VPg binds C-terminal region of cap-bound initiation factor 4E orthologue without exhibiting host cellular specificity.

To investigate the binding specificity of turnip mosaic virus (TuMV) viral protein-genome linked (VPg) with translation initiation factor 4E, we evaluated here the kinetic parameters for the interactions of human eIF4E, Caenorhabditis elegans IFE-3 and IFE-5 and Arabidopsis eIFiso4E, by surface plasmon resonance (SPR). The results indicated that TuMV VPg does not show a binding preference for Arabidopsis eIFiso4E, even though it is from a host species whereas the other eIF4E orthologues are not. Surprisingly, the effect of m(7)GTP on both the rate constants and equilibrium binding constants for the interactions of VPg differed for the four eIF4E orthologues. In the case of eIFiso4E and IFE-3, m(7)GTP increased k(on), but for eIF4E and IFE-5, it decreased k(on). To provide insight into the structural basis for these differences in VPg binding, tertiary structures of the eIF4E orthologues were predicted on the basis of the previously determined crystal structure of m(7)GpppA-bound human eIF4E. The results suggested that in cap-bound eIF4E orthologues, the VPg binds to the C-terminal region, which constitutes one side of the entrance to the cap-binding pocket, whereas in the cap-free state, VPg binds to the widely opened cap-binding pocket and its surrounding region. The binding of VPg to the C-terminal region was confirmed by the SPR analyses of N- or C-terminal residues-deleted eIF4E orthologues.

Turnip mosaic virus VPg interacts with Arabidopsis thaliana eIF(iso)4E and inhibits in vitro translation.

The interaction between turnip mosaic virus (TuMV) viral protein linked to the genome (VPg) and Arabidopsis thaliana eukaryotic initiation factor (iso)4E (eIF(iso)4E) was investigated to address the influence of potyviral VPg on host cellular translational initiation. Affinity chromatographic analysis showed that the region comprising amino acids 62-70 of VPg is important for the interaction with eIF(iso)4E. In vitro translation analysis showed that the addition of VPg significantly inhibited translation of capped RNA in eIF(iso)4E-reconstituted wheat germ extract. This result indicates that VPg inhibits cap-dependent translational initiation via binding to eIF(iso)4E. The inhibition by VPg of in vitro translation of RNA with wheat germ extract did not depend on RNase activity. Our present results may indicate that excess VPg produced at the encapsidation stage shuts off cap-dependent translational initiation in host cells by inhibiting complex formation between eIF(iso)4E and cellular mRNAs.