Broad-spectrum Β-lactam Antibiotics Research Articles

Microbiological Epidemiology Depending on Time to Occurrence of Prosthetic Joint Infection (PJI): Impact on the Empirical Antimicrobial Strategies

Background Empirical antimicrobial therapy of prosthetic-joint infection (PJI) is a major clinical challenge and current guidelines recommend the combination of vancomycin plus a broad-spectrum β-lactamin. As Gram-negative bacilli (GNB) are probably less represented in late infections, we evaluate the microbiological epidemiology in patients with PJI according to the chronology of infection.Methods All patients managed in a reference center for complex bone and joint infections in France (2011 and 2016) were included in a prospective cohort study. Microbiological data at the time of diagnosis were collected and analyzed according to the chronology of infection.ResultsWe included 567 PJI (284 males, 50.1%; median age 70.3 years). The median occurrence time was 23.4 weeks after prosthesis implantation (285 hip and 255 knee PJI, which were revision prosthesis in 216 [40.3%] cases). Microbiological bone samples found 164 [28.9%] S. aureus (including 26 [16.3%] MRSA), 162 [28.6%] coagulase-negative Staphylococci (CoNS, including 80 [58.8%] methicillin-resistant CoNS), 80 (14.1%) Enterobacteriaceae, 74 (13.1%) Streptococci, and 85 (15.0%) anaerobes (including 60 [10.6%] Propionibacterium). Infection was plurimicrobial in 10 [18.2%] cases. Among the 183 patients (32%) with late PJI (occurring >1 year), obtained after exclusion of the 59 patients (10.4%) with hematogenous origins, Enterobacteriacecae (n = 8; 4.4%; P < 10–3) were much less represented than in patients with early PJI occurring <1 year. No difference was observed regarding the the presence of non-fermenting GNB, with a prevalence of 4.6 and 2.7% in early and late PJI, respectively. Taken together, these data suggest that a broad-spectrum β-lactam antibiotic might be useful in only 12 (6.6%) patients with late PJI, compared with 66 (20.3%) patients with early PJI (P < 10–3). Of note, there were statistically more anaerobes (n = 40; 21.9%) in late PJI, including 32 Propionibacterium (17.5%; P < 10–3).Conclusion Considering the minority amount of GNB in late post-operative PJI and the overrepresentation of anaerobes including P. acnes, the empirical treatment should be reconsidered, especially when a two-stage exchange is planned. In those situations, another acceptable option could be the vancomycin+clindamycin combination.Disclosures T. Ferry, HERAEUS: Consultant, Speaker honorarium. S. Lustig, Heraeus: Consultant, Consulting fee.

Phenotypic and molecular detection of the bla KPC gene in clinical isolates from inpatients at hospitals in São Luis, MA, Brazil.

BackgroundBacteria that produce Klebsiella pneumoniae carbapenemases (KPCs) are resistant to broad-spectrum β-lactam antibiotics. The objective of this study was to phenotypically and genotypically characterize the antibiotic susceptibility to carbapenems of 297 isolates recovered from clinical samples obtained from inpatients at 16 hospitals in São Luis (Maranhão, Brazil).MethodsThe study was conducted using phenotypic tests and molecular methods, including polymerase chain reaction (PCR), sequencing and enterobacterial repetitive intergenic consensus (ERIC)-PCR. The nonparametric chi-square test of independence was used to evaluate the associations between the bacterial bla KPC gene and the modified Hodge test, and the chi-square adherence test was used to assess the frequency of carbapenemases and their association with the bla KPC gene.ResultsThe most frequently isolated species were Acinetobacter baumannii (n = 128; 43.0%), K. pneumoniae (n = 75; 25.2%), and Pseudomonas aeruginosa (n = 42; 14.1%). Susceptibility assays showed that polymixin B was active against 89.3% of the bacterial isolates. The Acinetobacter spp. and K. pneumoniae strains were susceptible to amikacin and tigecycline, and Pseudomonas spp. were sensitive to gentamicin and amikacin. Among the 297 isolates, 100 (33.7%) were positive for the bla KPC gene, including non-fermentative bacteria (A. baumannii) and Enterobacteriaceae species. Among the isolates positive for the bla KPC gene, K. pneumoniae isolates had the highest positivity rate of 60.0%. The bla KPC gene variants detected included KPC-2, which was found in all isolates belonging to species of the Enterobacteriaceae family. KPC-2 and KPC-3 were observed in A. baumannii isolates. Importantly, the bla KPC gene was also detected in three Raoultella isolates and one isolate of the Pantoea genus. ERIC-PCR patterns showed a high level of genetic diversity among the bacterial isolates; it was capable of distinguishing 34 clones among 100 strains that were positive for bla KPC and were circulating in 11 of the surveyed hospitals.ConclusionsThe high frequency of the bla KPC gene and the high degree of clonal diversity among microorganisms isolated from patients from different hospitals in São Luis suggest the need to improve the quality of health care to reduce the incidence of infections and the emergence of carbapenem resistance in these bacteria as well as other Gram-negative pathogens.

Benzylpenicillin plus an aminoglycoside versus meropenem in neutropenic lymphoma and leukaemia patients with a suspected bacterial infection: a randomized, controlled trial

ObjectivesIn Norway, initial treatment of febrile neutropenia (FN) has traditionally been benzylpenicillin plus an aminoglycoside. Internationally, FN is often treated with a broad-spectrum β-lactam antibiotic. We aimed to compare these two regimens in a prospective, randomized, trial in patients with lymphoma or leukaemia with an expected period of neutropenia ≥7 days, and a suspected bacterial infection. MethodsAdult neutropenic patients with lymphoma or leukaemia, and a suspected bacterial infection, were randomized for treatment with benzylpenicillin plus an aminoglycoside or meropenem. The primary endpoint was clinical success, defined as no modification of antibiotics and clinical stability 72 h after randomization. ResultsAmong 322 randomized patients, 297 proved evaluable for analyses. Fifty-nine per cent (95% CI 51%–66%), (87/148) of the patients given benzylpenicillin plus an aminoglycoside were clinically stable, and had no antibiotic modifications 72 h after randomization, compared with 82% (95% CI 75%–87%), (122/149) of the patients given meropenem (p <0.001). When the antibiotic therapy was stopped, 24% (95% CI 18%–32%), (36/148) of the patients given benzylpenicillin plus an aminoglycoside, compared with 52% (95% CI 44%–60%), (78/149) of the patients given meropenem, had no modifications of their regimens (p <0.001). In the benzylpenicillin plus an aminoglycoside arm, the all-cause fatality within 30 days of randomization was 3.4% (95% CI 1.2%–7.9%), (5/148) of the patients, compared with 0% (95% CI 0.0%–3.0%), (0/149) of the patients in the meropenem arm (p 0.03). ConclusionClinical success was more common in FN patients randomized to meropenem compared with the patients randomized to benzylpenicillin plus an aminoglycoside. The all-cause fatality was higher among the patients given benzylpenicillin plus an aminoglycoside.

A microbiological method for determining serum levels of broad spectrum β-lactam antibiotics in critically ill patients

BackgroundRecent studies show that suboptimal blood levels of β-lactam antibiotics are present in intensive care unit (ICU) patients. A common reference method for assessing drug concentrations is liquid chromatography coupled with mass-spectrometry (LC-MS) which is highly accurate but rarely available outside reference centres. Thus, our aim was to develop a microbiological method for monitoring β-lactam antibiotic serum levels which could be used at any hospital with a microbiological laboratory. MethodsThe method was developed as a 96-well broth microdilution format to assess the concentrations of cefotaxime (CTX), meropenem (MER), and piperacillin (PIP). Patient serum containing antibiotics were diluted in suspensions of bacteria with known minimal inhibitory concentrations (MICs). Serum antibiotic concentrations were calculated by dividing the MIC with the dilution factor at which the serum inhibited growth of the bacterial suspension. Serum (n=88) from ICU patients at four hospitals in south-east Sweden were analysed and compared to LC-MS analysis. ResultsThe overall accuracy and precision for spiked samples and patient samples was within the pre-set target of ±20.0% for all drugs. There was a significant correlation between the microbiological assay and LC-MS for the patient samples (CTX: r=0.86, n=31; MER: r=0.96, n=11; PIP: r=0.88, n=39) and the agreement around the clinical cut-off for CTX (4.0mg/l), MER (2.0mg/l) and PIP (16.0mg/l) was 90%, 100% and 87%, respectively. ConclusionThe microbiological method has a performance for determination of serum levels of meropenem, piperacillin and cefotaxime suitable for clinical use. It is an inexpensive method applicable in any microbiology laboratory.

Discovery of potent wall teichoic acid early stage inhibitors

The widespread emergence of methicillin-resistant Staphylococcus aureus (MRSA) has dramatically eroded the efficacy of current β-lactam antibiotics and created an urgent need for novel treatment options. Using an S. aureus phenotypic screening strategy, we have identified small molecule early stage wall teichoic acid (WTA) pathway-specific inhibitors predicted to be chemically synergistic with β-lactams. These previously disclosed inhibitors, termed tarocins, demonstrate by genetic and biochemical means inhibition of TarO, the first step in WTA biosynthesis. Tarocins demonstrate potent bactericidal synergy in combination with broad spectrum β-lactam antibiotics across diverse clinical isolates of methicillin-resistant Staphylococci. The synthesis and structure–activity relationships (SAR) of a tarocin series will be detailed. Tarocins and other WTA inhibitors may provide a rational strategy to develop Gram-positive bactericidal β-lactam combination agents active against methicillin-resistant Staphylococci.

Serum β-lactam concentrations in critically ill patients with cirrhosis: a matched case-control study.

The pharmacokinetics of β-lactam antibiotics have not been well defined in critically ill patients with cirrhosis. We reviewed data from critically ill patients with cirrhosis and matched controls in whom routine therapeutic drug monitoring of two broad-spectrum β-lactam antibiotics (piperacillin/tazobactam and meropenem) had been performed. Serum drug concentrations were measured twice by high-performance liquid chromatography. Antibiotic pharmacokinetics were calculated using a one-compartment model. We considered that therapy was adequate when serum drug concentrations were between 4 and 8 times the minimal inhibitory concentration of Pseudomonas aeruginosa during optimal periods of time for each drug (≥ 50% for piperacillin/tazobactam; ≥ 40% for meropenem). We studied 38 patients with cirrhosis (16 for piperacillin/tazobactam and 22 for meropenem) and 38 matched controls. Drug dosing was similar in the two groups. The pharmacokinetics analysis showed a lower volume of distribution of meropenem (P = 0.05) and a lower antibiotic clearance of piperacillin/tazobactam (P = 0.009) in patients with cirrhosis than in the matched controls. Patients with cirrhosis were more likely than those without cirrhosis to have excessive serum β-lactam concentrations (P = 0.015), in particular for piperacillin/tazobactam. Standard β-lactam antibiotics regimens resulted in excessive serum concentrations in two thirds of the patients with cirrhosis. This was particularly true for piperacillin/tazobactam, probably because of reduced drug clearance.

Population pharmacokinetic modeling of cefadroxil renal transport in wild-type and Pept2 knockout mice

1. Cefadroxil is a broad-spectrum β-lactam antibiotic that is widely used in the treatment of various infectious diseases. Currently, poor understanding of the drug’s pharmacokinetic profiles and disposition mechanism(s) prevents determining optimal dosage regimens and achieving ideal antibacterial responses in patients. In the present retrospective study, we developed a population pharmacokinetic model of cefadroxil in wild-type and Pept2 knockout mice using the nonlinear mixed effect modeling (NONMEM) approach.2. Cefadroxil pharmacokinetics were best described by a two-compartment model, with both saturable and nonsaturable elimination processes to/from the central compartment. Through this modeling approach, pharmacokinetic parameters in wild-type and Pept2 knockout mice were well estimated, respectively, as follows: volume of central compartment V1 (3.43 versus 4.23 mL), volume of peripheral compartment V2 (5.98 versus 8.61 mL), intercompartment clearance Q (0.599 versus 0.586 mL/min) and linear elimination rate constant K10 (0.111 versus 0.070 min−1). Moreover, the secretion kinetics (i.e. Vm1 = 17.6 nmoL/min and Km1 = 37.1 µM) and reabsorption kinetics (i.e. Vm2 = 15.0 nmoL/min and Km2 = 27.1 µM) of cefadroxil were quantified in kidney, for the first time, under in vivo conditions.3. Our model provides a unique tool to quantitatively predict the dose-dependent nonlinear disposition of cefadroxil, as well as the potential for transporter-mediated drug interactions.

Caracterización de mecanismos de resistencia por biología molecular: Staphylococcus aureus resistente a meticilina, β-lactamasas de espectro extendido y carbapenemasas

Multi-drug resistance in bacterial pathogens increases morbidity and mortality in infected patients and it is a threat to public health concern by their high capacity to spread. For both reasons, the rapid detection of multi-drug resistant bacteria is critical. Standard microbiological procedures require 48-72 h to provide the antimicrobial susceptibility results, thus there is emerging interest in the development of rapid detection techniques. In recent years, the use of selective and differential culture-based methods has widely spread. However, the capacity for detecting antibiotic resistance genes and their low turnaround times has made molecular methods a reference for diagnosis of multidrug resistance. This review focusses on the molecular methods for detecting some mechanisms of antibiotic resistance with a high clinical and epidemiological impact: a) Enzymatic resistance to broad spectrum β-lactam antibiotics in Enterobacteriaceae, mainly extended spectrum β-lactamases (ESBL) and carbapenemases; and b) methicillin resistance in Staphylococcus aureus.

A Simulation Study Reveals Lack of Pharmacokinetic/Pharmacodynamic Target Attainment in De-escalated Antibiotic Therapy in Critically Ill Patients.

De-escalation of empirical antibiotic therapy is often included in antimicrobial stewardship programs in critically ill patients, but differences in target attainment when antibiotics are switched are rarely considered. The primary objective of this study was to compare the fractional target attainments of contemporary dosing of empirical broad-spectrum β-lactam antibiotics and narrower-spectrum antibiotics for a number pathogens for which de-escalation may be considered. The secondary objective was to determine whether alternative dosing strategies improve target attainment. We performed a simulation study using published population pharmacokinetic (PK) studies in critically ill patients for a number of broad-spectrum β-lactam antibiotics and narrower-spectrum antibiotics. Simulations were undertaken using a data set obtained from critically ill patients with sepsis without absolute renal failure (n = 49). The probability of target attainment of antibiotic therapy for different microorganisms for which de-escalation was applied was analyzed. EUCAST MIC distribution data were used to calculate fractional target attainment. The probability that therapeutic exposure will be achieved was lower for the narrower-spectrum antibiotics with conventional dosing than for the broad-spectrum alternatives and could drastically be improved with higher dosages and different modes of administrations. For a selection of microorganisms, the probability that therapeutic exposure will be achieved was overall lower for the narrower-spectrum antibiotics using conventional dosing than for the broad-spectrum antibiotics.

Combinations of β-Lactam Antibiotics Currently in Clinical Trials Are Efficacious in a DHP-I-Deficient Mouse Model of Tuberculosis Infection.

We report here a dehydropeptidase-deficient murine model of tuberculosis (TB) infection that is able to partially uncover the efficacy of marketed broad-spectrum β-lactam antibiotics alone and in combination. Reductions of up to 2 log CFU in the lungs of TB-infected mice after 8 days of treatment compared to untreated controls were obtained at blood drug concentrations and time above the MIC (T>MIC) below clinically achievable levels in humans. These findings provide evidence supporting the potential of β-lactams as safe and mycobactericidal components of new combination regimens against TB with or without resistance to currently used drugs.

Evaluation of Anti-bacterial and Anti-quorum Sensing Potential of Essential Oils Extracted by Supercritical CO2 Method Against Pseudomonas aeruginosa

Pseudomonas aeruginosa is an important opportunistic pathogen causing widespread infections by numerous virulence factors. The present investigation was to evaluate the antimicrobial and anti-quorum sensing (anti-QS) activities of essential oils (EOs) from Cuminum cyminum, Zingiber officinale, Myristica fragrans, Syzygium aromaticum, Cinnamomum verum, Rosmarinus officinalis, and Curcuma longa against the wild strain of Chromobacterium violaceum CV12472, clinical strain of P. aeruginosa (MM-1) and P. aeruginosa PAO1. All the EOs have shown antimicrobial activity against CV12472. Among these, five EOs namely C. cyminum, S. aromaticum, C. verum, R. officinalis and C. longa have shown anti-QS activity as evidenced by inhibition of violacein production. S. aromaticum and C. verum have shown promising anti-QS activity at very low concentration of 0.006 %, v/v and 0.01 %, v/v respectively. The study also analysed against the PAO1 and P. aeruginosa (MM-1). The MM-1 strain was resistant to the tested broad-spectrum β-lactam and cephalosporin antibiotics. Among the seven EOs, MICs of S. aromaticum and C. verum were found to be 0.78 %, v/v and also they have shown anti-QS activities at 0.39 %, v/v and 0.097 %, v/v as confirmed by the decreased pyocyanin production in the MM-1 strain. C. verum and S. aeromaticum inhibited the swarming motility of the tested P. aeruginosa strains. The present study suggested that the active principle from essential oils can be used as efficient herbal medicine for blocking the QS against the resistant clinical strains of P. aeruginosa.

β-lactam antibiotic concentrations during continuous renal replacement therapy.

IntroductionThe use of standard doses of β-lactam antibiotics during continuous renal replacement therapy (CRRT) may result in inadequate serum concentrations. The aim of this study was to evaluate the adequacy of unadjusted drug regimens (i.e., similar to those used in patients with normal renal function) in patients treated with CRRT and the influence of CRRT intensity on drug clearance.MethodsWe reviewed data from 50 consecutive adult patients admitted to our Department of Intensive Care in whom routine therapeutic drug monitoring (TDM) of broad-spectrum β-lactam antibiotics (ceftazidime or cefepime, CEF; piperacillin/tazobactam; TZP; meropenem, MEM) was performed using unadjusted β-lactam antibiotics regimens (CEF = 2 g q8h; TZP = 4 g q6h; MEM = 1 g q8h). Serum drug concentrations were measured twice during the elimination phase by high-performance liquid chromatography (HPLC-UV). We considered therapy was adequate when serum drug concentrations were between 4 and 8 times the minimal inhibitory concentration (MIC) of Pseudomonas aeruginosa during optimal periods of time for each drug (≥70% for CEF; ≥ 50% for TZP; ≥ 40% for MEM). Therapy was considered as early (ET) or late (LT) phase if TDM was performed within 48 hours of antibiotic initiation or later on, respectively.ResultsWe collected 73 serum samples from 50 patients (age 58 ± 13 years; Acute Physiology and Chronic Health Evaluation II (APACHE II) score on admission 21 (17–25)), 35 during ET and 38 during LT. Drug concentrations were above 4 times the MIC in 63 (90%), but above 8 times the MIC in 39 (53%) samples. The proportions of patients with adequate drug concentrations during ET and LT were quite similar. We found a weak but significant correlation between β-lactam antibiotics clearance and CRRT intensity.ConclusionsIn septic patients undergoing CRRT, doses of β-lactam antibiotics similar to those given to patients with normal renal function achieved drug levels above the target threshold in 90% of samples. Nevertheless, 53% of samples were associated with very high drug levels and daily drug regimens may need to be adapted accordingly.

Computational and functional analysis of β-lactam resistance in Zymomonas mobilis

Zymomonas mobilis, a Gram-negative ethanologenic non-pathogenic bacterium, is reported to exhibit resistance to high concentrations of β-lactam antibiotics. In the present study, Z. mobilis was found to be resistant to I-IV generations of cephalosporins and carbapenems, i.e. narrow, broad and extended spectrum β-lactam antibiotics. We have analysed the genome of Z. mobilis (GenBank accession No.: NC 006526) harbouring multiple genes coding for β-lactamases (BLA), β-lactamase domain containing proteins (BDP) and penicillin binding proteins (PBP). The conserved domain database analysis of BDPs predicted them to be members of metallo β-lactamase superfamily. Further, class C specific multidomain AmpC (β-lactamase C) was found in the three β-lactamases. The β-lactam resistance determinants motifs, HXHXD, KXG, SXXK, SXN, and YXN are present in the BLAs, BDPs and PBPs of Z. mobilis. The predicted theoretical pI and aliphatic index values suggested their stability. One of the PBPs, PBP2, was predicted to share functional association with rod shape determining proteins (GenBank accession Nos. YP_162095 and YP_162091). Homology modelling of three dimensional structures of the β-lactam resistance determinants and further docking studies with penicillin and other β-lactam antibiotics indicated their substrate-specificity. Semi-quantitative PCR analysis indicated that the expression of all BLAs and one BDP are induced by penicillin. Disk diffusion assay, SDS-PAGE and zymogram analysis confirms the substrate specificity of the β-lactam resistance determinants. This study gives a broader picture of the β-lactam resistance determinants of a non-pathogenic ethanologenic Z. mobilis bacterium that could have implications in laboratories since it is routinely used in many research laboratories in the world for ethanol, fructooligosaccharides, levan production and has also been reported to be present in wine and beer as a spoilage organism.

Synthesis, pH-dependent, and plasma stability of meropenem prodrugs for potential use against drug-resistant tuberculosis

Meropenem, a broad-spectrum parenteral β-lactam antibiotic, in combination with clavulanate has recently shown efficacy in patients with extensively drug-resistant tuberculosis. As a result of meropenem’s short half-life and lack of oral bioavailability, the development of an oral therapy is warranted for TB treatment in underserved countries where chronic parenteral therapy is impractical. To improve the oral absorption of meropenem, several alkyloxycarbonyloxyalkyl ester prodrugs with increased lipophilicity were synthesized and their stability in physiological aqueous solutions and guinea pig as well as human plasma was evaluated. The stability of prodrugs in aqueous solution at pH 6.0 and 7.4 was significantly dependent on the ester promoiety with the major degradation product identified as the parent compound meropenem. However, in simulated gastrointestinal fluid (pH 1.2) the major degradation product identified was ring-opened meropenem with the promoiety still intact, suggesting the gastrointestinal environment may reduce the absorption of meropenem prodrugs in vivo unless administered as an enteric-coated formulation. Additionally, the stability of the most aqueous stable prodrugs in guinea pig or human plasma was short, implying a rapid release of parent meropenem.

Clinical Significance of Infections Caused by Extended-Spectrum β-Lactamase-ProducingEnterobacteriaceaeBlood Isolates with Inducible AmpC β-Lactamase

Few studies have investigated the clinical features and outcomes for extended-spectrum β-lactamase (ESBL)-producing Enterobacter spp., Citrobacter spp., Serratia spp., and Morganella morganii (ECSM) bloodstream infections. This study was performed to investigate the clinical features and outcomes for ESBL-producing ECSM bloodstream infections. Patients with ECSM bloodstream infection were enrolled from October 2006 to March 2008. Of 124 patients with ECSM bacteremia, 30 cases (24.2%) were ESBL-producing isolates. Immunosuppressive drugs use within 30 days (p=0.028), indwelling device at the time of bacteremia (p=0.042) and antibiotics use within 3 months (p=0.022) were independently associated with ESBL production in multivariate analysis. Overall 30-day mortality rate was 19.4% (24/124). When the 30-day mortality rate was evaluated, no significant difference was found between the ESBL group (16.6%; 5/30) and non-ESBL group (20.2%; 19/94). Hospitalization was longer in the ESBL group than in the non-ESBL group (65.4±92.8 vs. 32.9±37.8 days, respectively; p=0.007). The recent use of antibiotics (especially broad-spectrum cephalosporins and other β-lactam antibiotics) was an important risk factor for ESBL among ECSM bacteremia. ESBL production of ECSM isolates was not significantly associated with mortality but ESBL-producing organisms have an important impact on the duration of hospital stay and subsequent medical cost.

Critical appraisal of ceftaroline in the management of community-acquired bacterial pneumonia and skin infections.

Ceftaroline is a novel broad-spectrum cephalosporin β-lactam antibiotic with activity against methicillin-resistant Staphylococcus aureus (MRSA) as well as multidrug-resistant Streptococcus pneumoniae among other routine Gram positive and Gram negative organisms. It has been approved by the US Food and Drug Administration for treatment of community-acquired bacterial pneumonia and acute bacterial skin and skin structure infections (ABSSSIs). Ceftaroline is approved for treatment of ABSSSI due to MRSA, however currently there are no data for pneumonia due to MRSA in humans. Herein we review the major clinical trials as well as ceftaroline microbiology, pharmacokinetics, and safety, followed by a look at further directions for investigation of this new agent.

Providing β-Lactams a Helping Hand: Targeting the Ampc β-Lactamase Induction Pathway

A major cause of the clinical failure of broad-spectrum β-lactam antibiotics against Pseudomonas aeruginosa and many Enterobacteriaceae species are chromosomal mutations that lead to the hyperproduction of AmpC β-lactamase. These mutations typically affect proteins within the peptidoglycan (PG) recycling pathway, as well as proteins that are modulated by metabolic intermediates of this pathway. Blocking PG recycling and associated sensing mechanisms with small-molecule inhibitors holds promise as a strategy for overcoming AmpC-mediated resistance that results from the selection of mutations during β-lactam therapy, or from the direct acquisition of infections by AmpC-producing mutants. Here we report on the structural and functional biology of potential drug targets within the Gram-negative PG recycling pathway and the utility of blocking PG recycling as a means of attenuating AmpC-mediated resistance in P. aeruginosa.

The Footprints of Gut Microbial–Mammalian Co-Metabolism

Gut microbiota are associated with essential various biological functions in humans through a "network" of microbial-host co-metabolism to process nutrients and drugs and modulate the activities of multiple pathways in organ systems that are linked to different diseases. The microbiome impacts strongly on the metabolic phenotypes of the host, and hence, metabolic readouts can give insights into functional metagenomic activity. We applied an untargeted mass spectrometry (MS) based metabonomics approach to profile normal Wistar rats exposed to a broad spectrum β-lactam antibiotic imipenem/cilastatin sodium, at 50 mg/kg/daily for 4 days followed by a 14-day recovery period. In-depth metabolic phenotyping allowed identification of a panel of 202 urinary and 223 fecal metabolites significantly related to end points of a functional metagenome (p < 0.05 in at least one day), many of which have not been previously reported such as oligopeptides and carbohydrates. This study shows extensive gut microbiota modulation of host systemic metabolism involving short-chain fatty acids, tryptophan, tyrosine metabolism, and possibly a compensatory mechanism of indole-melatonin production. Given the integral nature of the mammalian genome and metagenome, this panel of metabolites will provide a new platform for potential therapeutic markers and mechanistic solutions to complex problems commonly encountered in pathology, toxicology, or drug metabolism studies.

Contribution of BlaA and BlaB β-Lactamases to Antibiotic Susceptibility of Yersinia enterocolitica Biovar 1B

Highly pathogenic Yersinia enterocolitica biovar 1B produces two distinct beta-lactamases, BlaA and BlaB. Mutants of a representative biovar 1B isolate were constructed and evaluated to determine the extent of limitation of susceptibility to broad-spectrum beta-lactam antibiotics by BlaA and BlaB. The results demonstrated that BlaA, a class A enzyme, plays a significant role in limiting susceptibility to penicillins and cephalosporins. The contribution of BlaB, a class C enzyme, was less profound and was limited primarily to cephalosporin susceptibility.

Cluster of bloodstream infections caused by KPC-2 carbapenemase-producing Klebsiella pneumoniae in Manhattan

Carbapenems are considered the agents of choice for treatment of serious infections caused by resistant gram-negative organisms. A new group of class A beta-lactamases, known as KPC-type carbapenemases, has recently been described and poses a serious clinical challenge. Seven patients with bloodstream infections caused by Klebsiella pneumoniae isolates with decreased susceptibility to carbapenems were identified between January and April 2005 in the intensive care units of a hospital in Manhattan. Isolate identification and susceptibility testing were performed according to National Committee for Clinical Laboratory Standards methodology. All isolates were ribotyped and screened for (bla)KPC by polymerase chain reaction. The polymerase chain reaction product underwent nucleotide sequencing for one of the isolates. Medical records were reviewed retrospectively. Six isolates were carbapenem-resistant with minimum inhibitory concentrations for imipenem of >8microg/mL. Ribotyping showed that all isolates belonged to a single clone. All isolates possessed (bla)KPC and nucleotide sequencing identified the allelic type KPC-2. Patients' median age was 68 years. The median duration of hospitalization was 25.5 days before the first positive blood culture. Five of 6 patients received previous broad-spectrum beta-lactam antibiotics but none received prior carbapenems. Five of 6 isolates were susceptible to polymyxin B. Three of the 5 patients were treated with polymyxin B and 1 survived. Overall, only 2 of the 6 patients survived. This report describes the first outbreak of KPC-2 carbapenemase-producing K pneumoniae bloodstream infections in a hospital in Manhattan.