Pharmacokinetics, Tissue Distribution, and Efficacy of VIO-001 (Meropenem/Piperacillin/Tazobactam) for Treatment of Methicillin-Resistant Staphylococcus aureus Bacteremia in Immunocompetent Rabbits with Chronic Indwelling Vascular Catheters.

Methicillin-resistant Staphylococcus aureus (MRSA) infections of surgically implanted subcutaneous vascular catheters (SISVCs) cause serious morbidity in patients with chronic illnesses. Previous in vitro and murine models demonstrated the synergistic interaction of equimolar concentrations of meropenem/piperacillin/tazobactam (MPT) (VIO-001) against MRSA infection. We investigated the pharmacokinetics (PK) and efficacy of VIO-001 for the treatment of MRSA bacteremia in immunocompetent rabbits with SISVCs. In PK studies, we determined that optimal dosing to achieve a time above 4× MIC (T>4×MIC) of a duration of 3 to 3.30 h required a 1-h infusion with every-4-h (Q4h) dosing. Study groups in efficacy experiments consisted of MPT combinations of 100/150/100 mg/kg of body weight (MPT100), 200/300/200 mg/kg (MPT200), and 400/600/400 mg/kg (MPT400); vancomycin (VAN) at 15 mg/kg; and untreated controls (UC). The inoculum of MRSA isolate USA300-TCH1516 (1 × 103 organisms) was administered via the SISCV on day 1 and locked for 24 h. The 8-day therapy started at 24 h postinoculation. There was a significant reduction of MRSA in blood cultures from the SISVCs in all treatment groups, with full clearance on day 4, versus UCs (P < 0.05). Consistent with the clearance of SISVC-related infection, full eradication of MRSA was achieved in lungs, heart, liver, spleen, and kidneys at the end of the study versus UC (P < 0.01). These results strongly correlated with time-kill data, where MPT in the range of 4/6/4 µg/ml to 32/48/32 µg/ml demonstrated a significant 6-log decrease in the bacterial burden versus UC (P < 0.01). In summary, VIO-001 demonstrated a favorable PK/pharmacodynamic (PD) profile and activity against SISCV MRSA infection, bacteremia, and disseminated infection. This rabbit model provides a new system for understanding new antimicrobial agents against MRSA SISVC-related infection, and these data provide a basis for future clinical investigation.

Pharmacokinetics of Cefazolin and Vancomycin in Infants Undergoing Open-Heart Surgery With Cardiopulmonary Bypass.

BACKGROUND: Gram-positive bacteria account for nearly three-quarters of all surgical site infections. Antibiotic prophylaxis against these bacteria with cephalosporins or, in select circumstances, with vancomycin is considered standard of care for prevention of surgical site infections. There is little evidence to describe the optimal dosing regimen for surgical site infection prophylaxis in infants undergoing cardiac surgery, and a great deal of institutional variability exists in dosing prophylactic antibiotics. We designed this study to describe an optimal dose regimen for cephalosporin and vancomycin based on pharmacokinetic evidence for infant open-heart surgery on cardiopulmonary bypass. METHODS: Two separate cohorts of infants undergoing cardiac surgery with cardiopulmonary bypass were evaluated. Plasma concentrations of vancomycin (cohort 1, N = 10) and cefazolin (cohort 2, N = 10) were measured, and mixed-effects pharmacokinetic models were constructed for each drug. Simulations of various dosing regimens were performed to describe an appropriate dosing regimen necessary to maintain antibiotic concentrations above the susceptibility cutoff for staphylococci. RESULTS: Both cefazolin and vancomycin plasma concentration versus time profiles were characterized by a 2-compartment model. Subject weight was a significant covariate for V1 for vancomycin. Subject age was a significant covariate for V1 for cefazolin. Cardiopulmonary bypass did not influence concentration versus time profiles. Simulations demonstrated that a 1-hour vancomycin infusion (15 mg·kg), repeated every 12 hours and a 10-minute infusion of cefazolin (30 mg·kg), repeated every 4 hours maintained plasma concentrations above 4 µg·mL and 16 µg·mL, for vancomycin and cefazolin, respectively. Both concentrations are above the minimum inhibitory concentration 90 for most susceptible staphylococci. CONCLUSIONS: Prophylactic treatment of vancomycin 15 mg·kg infused >1 hour with 12-hour redosing and cefazolin 30 mg·kg infused >10 minutes with 4-hour redosing will maintain serum levels of each antibiotic above the susceptibility cut-offs for susceptible staphylococci in infants undergoing cardiac surgery. Cefazolin levels may be adequate for some, but not all, Gram-negative bacteria. The effect of cardiopulmonary bypass on pharmacokinetics is negligible.

Subcutaneous cefazolin to reduce surgical site infections in a porcine model.

BACKGROUND: Surgical site infections (SSIs) pose a significant health and financial burden. A key aspect of appropriate prophylaxis is the administration of antibiotics intravenously (IV). However, subcutaneous administration of antibiotics is not well described in the literature. During surgery, we hypothesize that subcutaneous injection may provide better protection against SSIs. To better understand the kinetics after subcutaneous injection, we describe the serum concentrations of cefazolin in a porcine model. MATERIALS AND METHODS: Juvenile mini-Yucatan pigs were administered 20 mL of 25 mg/kg cefazolin subcutaneously, and serial blood samples were taken for 3 h. Blood samples were analyzed for cefazolin concentration using chromatography. Pharmacokinetic data were calculated based on the blood serum concentrations. RESULTS: Maximum serum concentrations of cefazolin were achieved 42.6 ± 2.0 min after the time of injection and were found to be 18.8 ± 7.4 µg/mL. The elimination rate constant was 0.0033 ± 0.0016 min-1 and the half-life was 266 ± 149 min. The area under the curve was 4940 ± 1030 µg × min/mL. The relative bioavailability of subcutaneous injection was 95% +5%/-20%. CONCLUSIONS: Subcutaneous administration of cefazolin achieves a significantly lower maximum serum concentration than IV injection. As a result, higher doses of antibiotic can be injected locally without incurring systemic toxicity. Subcutaneous administration will therefore result in higher concentrations of antibiotic for a longer time at the incision site compared with standard IV administration. This strategy of antibiotic delivery may be more effective in preventing SSIs. Further studies are needed to detail the exact effect of subcutaneous antibiotic injection on SSI rates.

Multicenter Evaluation of Ceftazidime-Avibactam and Ceftolozane-Tazobactam Inhibitory Activity against Meropenem-Nonsusceptible Pseudomonas aeruginosa from Blood, Respiratory Tract, and Wounds.

The recent escalation of occurrences of carbapenem-resistant Pseudomonas aeruginosa has been recognized globally and threatens to erode the widespread clinical utility of the carbapenem class of compounds for this prevalent health care-associated pathogen. Here, we compared the in vitro inhibitory activity of ceftazidime-avibactam and ceftolozane-tazobactam against 290 meropenem-nonsusceptible Pseudomonas aeruginosa nonduplicate clinical isolates from 34 U.S. hospitals using reference broth microdilution methods. Ceftazidime-avibactam and ceftolozane-tazobactam were active, with ceftolozane-tazobactam having significantly higher inhibitory activity than ceftazidime-avibactam. The heightened inhibitory activity of ceftolozane-tazobactam was sustained when the site of origin (respiratory, blood, or wound) and nonsusceptibility to other ß-lactam antimicrobials was considered. An extensive genotypic search for enzymatically driven ß-lactam resistance mechanisms revealed the exclusive presence of the VIM metallo-ß-lactamase among only 4% of the subset of isolates nonsusceptible to ceftazidime-avibactam, ceftolozane-tazobactam, or both. These findings suggest an important role for both ceftazidime-avibactam and ceftolozane-tazobactam against carbapenem-nonsusceptible Pseudomonas aeruginosa Further in vitro and in vivo studies are needed to better define the clinical utility of these novel therapies against the increasingly prevalent threat of multidrug-resistant Pseudomonas aeruginosa.

In vitro potency of amikacin and comparators against E. coli, K. pneumoniae and P. aeruginosa respiratory and blood isolates.

BACKGROUND: The purpose of this study was to define the potency of amikacin and comparator agents against a collection of blood and respiratory nosocomial isolates implicated in ICU based pulmonary infections gathered from US hospitals. METHODS: Minimum inhibitory concentrations of amikacin, aztreonam, cefepime, ceftazidime, ceftolozane/tazobactam, ceftriaxone, ciprofloxacin, imipenem, meropenem, piperacillin/tazobactam and tobramycin were tested against 2460 Gram-negative isolates. Amikacin had 96 % susceptibility against the combined E. coli and K. pneumoniae isolates and 95 % susceptibility against P. aeruginosa. RESULTS: Ninety-six percent of all of isolates tested were susceptible (i.e., MICs ≤16 mg/L) to amikacin by current laboratory standards which demonstrates a high level of activity to combat infections caused by these organisms including ESBL, MDR, ß-lactam and fluoroquinolone resistant strains. Moreover, 99 % of all organisms had amikacin MICs ≤64 mg/L. CONCLUSIONS: Overall, these data highlight the continued potency of amikacin and suggest that the achievable lung concentrations of approximately 5000 mg/L with the administration of the amikacin by inhalation (Amikacin Inhale, BAY41-6551) will exceed the MICs typically observed for P. aeruginosa, E. coli and K. pneumoniae in the hospital setting.

Impact of revised cefepime CLSI breakpoints on Escherichia coli and Klebsiella pneumoniae susceptibility and potential impact if applied to Pseudomonas aeruginosa.

The CLSI reduced the cefepime Enterobacteriaceae susceptibility breakpoint and introduced the susceptible-dose-dependent (S-DD) category. In this study, MICs were determined for a Gram-negative collection to assess the impact of this change. For Enterobacteriaceae, this resulted in <2% reduction in susceptibility, with 1% being S-DD. If applied to Pseudomonas aeruginosa, the % susceptibility (%S) dropped from 77% to 43%, with 34% being S-DD. The new breakpoints did little to the Enterobacteriaceae %S, but for P. aeruginosa, a profound reduction was seen in %S. The recognition of a S-DD response to cefepime should alert clinicians to the possible need for higher doses.

To add or not to add polysorbate 80: impact on colistin MICs for clinical strains of Enterobacteriaceae and Pseudomonas aeruginosa and quality controls.

Colistin adherence to plastics can be diminished by adding a surfactant, i.e., polysorbate 80. Incorporating polysorbate 80 resulted in 3 twofold and 2 twofold modal MIC decreases for Enterobacteriaceae and Pseudomonas aeruginosa, respectively. The reproducibility of the quality controls (QCs) with and without polysorbate 80 supports the use of Escherichia coli strain 25922 and P. aeruginosa strain 27853.

Comparison of 2 blood culture media shows significant differences in bacterial recovery for patients on antimicrobial therapy.

BACKGROUND: Antimicrobial removal devices in blood culture media are designed to remove antibiotics from the blood culture solution, thereby facilitating bacterial growth. How well these devices function clinically has not been established. METHODS: All blood drawn for culture from adult inpatients and emergency department visitors in a level I trauma center was placed in paired BACTEC Plus and BacT/Alert FAN culture media and studied simultaneously, consecutively, and prospectively between 1 February and 30 September 2011. All cultures were processed per standard laboratory protocols. RESULTS: Of 9395 total cultures collected, 1219 (13%) were positive, 831 were included, and 524 (33%) contained pathogens. BACTEC had a 4.5-hour faster detection time (P < .0001), and isolated exclusively 182 of 524 (35%; P < .001) pathogens, 136 of 345 (39%) of the gram-positive cocci (P < .001), 48 of 175 (27%; P = .02) of the gram-negative rods, 101 of 195 (52%) of Staphylococcus aureus (P < .001), and 59 of 120 (49%; P = .004) septic events. If active antibiotics had been dosed 0-4 or 4-48 hours prior to culture collection, the odds of that culture growing in BACTEC were 4.8- and 5.2-fold greater, respectively, than of growing in BacT/Alert (P < .0001). Both were equivalent in the recovery of yeast and when no antimicrobials were dosed. CONCLUSIONS: BACTEC media has faster time to detection and increased bacterial recovery over the BacT/Alert media in the following categories: overall growth, pathogens, septic events, gram-positive cocci, gram-negative rods, Staphylococcus aureus, and cultures where antimicrobials were dosed up to 48 hours before culture collection.

An Automated, Open-Source Workflow for the Generation of (3D) Fragment Libraries.

The recent success of fragment-based drug discovery (FBDD) is inextricably linked to adequate library design. To guide the design of our fragment libraries, we have constructed an automated workflow in the open-source KNIME software. The workflow considers chemical diversity and novelty of the fragments, and can also take into account the three-dimensional (3D) character. This design tool can be used to create large and diverse libraries but also to select a small number of representative compounds as a focused set of unique screening compounds to enrich existing fragment libraries. To illustrate the procedures, the design and synthesis of a 10-membered focused library is reported based on the cyclopropane scaffold, which is underrepresented in our existing fragment screening library. Analysis of the focused compound set indicates significant shape diversity and a favorable overall physicochemical profile. By virtue of its modular setup, the workflow can be readily adjusted to design libraries that focus on properties other than 3D shape.

In vitro evaluation of novel compounds against selected resistant Pseudomonas aeruginosa isolates.

We describe the activities of RX-P763, RX-P766, RX-P770, RX-P792, RX-P793, and RX-P808 against strains of resistant Pseudomonas aeruginosa. These compounds target the large subunit of the bacterial ribosome and have broad-spectrum activities against multidrug-resistant pathogens. All compounds demonstrated in vitro activity against P. aeruginosa, with MIC(90) values of 4 to 8 µg/ml (range, 0.5 to 64). These novel compounds had narrow MIC distributions and maintained activity despite resistance phenotypes to other commonly utilized agents.

In vivo comparison of CXA-101 (FR264205) with and without tazobactam versus piperacillin-tazobactam using human simulated exposures against phenotypically diverse gram-negative organisms.

CXA-101 is a novel antipseudomonal cephalosporin with enhanced activity against Gram-negative organisms displaying various resistance mechanisms. This study evaluates the efficacy of exposures approximating human percent free time above the MIC (%fT > MIC) of CXA-101 with or without tazobactam and piperacillin-tazobactam (TZP) against target Gram-negative organisms, including those expressing extended-spectrum ß-lactamases (ESBLs). Sixteen clinical Gram-negative isolates (6 Pseudomonas aeruginosa isolates [piperacillin-tazobactam MIC range, 8 to 64 µg/ml], 4 Escherichia coli isolates (2 ESBL and 2 non-ESBL expressing), and 4 Klebsiella pneumoniae isolates (3 ESBL and 1 non-ESBL expressing) were used in an immunocompetent murine thigh infection model. After infection, groups of mice were administered doses of CXA-101 with or without tazobactam (2:1) designed to approximate the %fT > MIC observed in humans given 1 g of CXA-101 with or without tazobactam every 8 h as a 1-h infusion. As a comparison, groups of mice were administered piperacillin-tazobactam doses designed to approximate the %fT > MIC observed in humans given 4.5 g piperacillin-tazobactam every 6 h as a 30-min infusion. Predicted piperacillin-tazobactam %fT > MIC exposures of greater than 40% resulted in static to >1 log decreases in CFU in non-ESBL-expressing organisms with MICs of ≤32 µg/ml after 24 h of therapy. Predicted CXA-101 with or without tazobactam %fT > MIC exposures of ≥37.5% resulted in 1- to 3-log-unit decreases in CFU in non-ESBL-expressing organisms, with MICs of ≤16 µg/ml after 24 h of therapy. With regard to the ESBL-expressing organisms, the inhibitor combinations showed enhanced CFU decreases versus CXA-101 alone. Due to enhanced in vitro potency and resultant increased in vivo exposure, CXA-101 produced statistically significant reductions in CFU in 9 isolates compared with piperacillin-tazobactam. The addition of tazobactam to CXA-101 produced significant reductions in CFU for 7 isolates compared with piperacillin-tazobactam. Overall, human simulated exposures of CXA-101 with or without tazobactam demonstrated improved efficacy versus piperacillin-tazobactam.

Bronchopulmonary pharmacokinetic and pharmacodynamic profiles of levofloxacin 750 mg once daily in adults undergoing treatment for acute exacerbation of chronic bronchitis.

While previous reports have described the bronchopulmonary profile of the fluoroquinolones in healthy volunteers, limited data are available in infected patients. The purpose of this study was to determine the intrapulmonary profile of high-dose (750 mg) levofloxacin in patients during an acute exacerbation of chronic bronchitis (AECB). Twenty-four patients experiencing clinical signs and symptoms of AECB were enrolled. Once enrolled, patients received levofloxacin 750 mg once daily × 5 days. Bronchoalveolar lavage aspirates and simultaneous plasma samples were obtained at 4 h, 12 h or 24 h after the third dose. Concentrations in biologic matrixes were determined with a validated HPLC method. Epithelial lining fluid (ELF) concentrations were calculated using the urea dilution method. Five patients did not complete the trial, 19 patients underwent bronchoscopy, 18 (52 ± 13 yrs) had sufficient samples for analysis and confirmed medication compliance. Mean plasma concentrations at 4, 12, and 24 h were 8.0 ± 2.5, 5.8 ± 1.2, and 2.2 ± 1.2 µg/mL. Mean ELF values at 4, 12, and 24 h were 7.5 ± 3.0, 8.3 ± 6.0, and 1.2 ± 0.9 µg/mL. Mean alveolar macrophage (AM) concentrations at 4, 12, and 24 h were 38.5 ± 43.7, 13.4 ± 14.4, and 9.0 ± 7.5 µg/mL. The penetration (ELF/plasma ratio) into the infection site was 113%. In these subjects with AECB, levofloxacin 750 mg once daily reached adequate exposures in the plasma, ELF, and AMs for the most commonly associated pathogens.

The Value and Institutional Impact of an In-System Laboratory Testing During the COVID-19 Pandemic.

In-system clinical laboratories have proven themselves to be a fundamentally important resource to their institutions during the COVID-19 pandemic of the past year. The ability to provide SARS-CoV-2 molecular testing to our hospital system allowed us to offer the best possible care to our patients, and to support neighboring hospitals and nursing homes. In-house testing led to significant revenue enhancement to the laboratory and institution, and attracted new patients to the system. Timely testing of inpatients allowed the majority who did not have COVID-19 infection to be removed from respiratory and contact isolation, conserving valuable personal protective equipment and staff resources at a time that both were in short supply. As 2020 evolved and our institution restarted delivery of routine care, the availability of in-system laboratory testing to deliver both accurate and timely results was absolutely critical. In this article, we attempt to demonstrate the value and impact of an in-system laboratory during the COVID-19 pandemic. A strong in-house laboratory service was absolutely critical to institutional operational and financial success during 2020, and will ensure resiliency in the future as well.

Tissue penetration and pharmacokinetics of tigecycline in diabetic patients with chronic wound infections described by using in vivo microdialysis.

Tissue penetration of systemic antibiotics is an important consideration for positive outcomes in diabetic patients. Herein we describe the exposure profile and penetration of tigecycline in the interstitial fluid of wound margins versus that of uninfected thigh tissue in 8 adult diabetic patients intravenously (IV) administered 100 mg and then 50 mg of tigecycline twice daily for 3 to 5 doses. Prior to administration of the first dose, 2 microdialysis catheters were inserted into the subcutaneous tissue, the first within 10 cm of the wound margin and the second in the thigh of the same extremity. Samples for determination of plasma and tissue concentrations were simultaneously collected over 12 h under steady-state conditions. Tissue concentrations were corrected for percent in vivo recovery by the retrodialysis technique. Plasma samples were also collected for determination of protein binding at 1, 6, and 12 h postdose for each patient. Protein binding data were corrected using a fitted polynomial equation. The mean patient weight was 95.1 kg (range, 63.6 to 149.2 kg), the mean patient age was 63.5 ± 9.4 years, and 75% of the patients were males. The mean values for the plasma, thigh, and wound free area under the concentration-time curve from 0 to 24 h (fAUC(0-24)) were 2.65 ± 0.33, 2.52 ± 1.15, and 2.60 ± 1.02 µg·h/ml, respectively. Protein binding was nonlinear, with the percentage of free drug increasing with decreasing serum concentrations. Exposure values for thigh tissue and wound tissue were similar (P = 0.986). Mean steady-state tissue concentrations for the thigh and wound were similar at 0.12 ± 0.02 µg/ml, and clearance from the tissues appeared similar to that from plasma. Tissue penetration ratios (tissue fAUC/plasma fAUC) were 99% in the thigh and 100% in the wound (P = 0.964). Tigecycline penetrated equally well into wound and uninfected tissue of the same extremity.

Comparison of the activity of a human simulated, high-dose, prolonged infusion of meropenem against Klebsiella pneumoniae producing the KPC carbapenemase versus that against Pseudomonas aeruginosa in an in vitro pharmacodynamic model.

We have previously demonstrated that a high-dose, prolonged-infusion meropenem regimen (2 g every 8 h [q8h]; 3-hour infusion) can achieve 40% free drug concentration above the MIC against Pseudomonas aeruginosa with MICs of or=3 log CFU reduction against all KPC isolates within 6 h, followed by regrowth in all but two isolates. The targeted %fT>MIC (percent time that free drug concentrations remain above the MIC) exposure was achieved against both of these KPC isolates (100% fT>MIC versus MIC=2 microg/ml, 75% fT>MIC versus MIC=8 microg/ml). Against KPC isolates with MICs of 8 and 16 microg/ml that did regrow, actual meropenem exposures were significantly lower than targeted due to rapid in vitro hydrolysis, whereby targeted %fT>MIC was reduced with each subsequent dosing. In contrast, a >or=3 log CFU reduction was maintained over 24 h for all Pseudomonas isolates with meropenem MICs of 8 and 16 microg/ml. Although KPC and P. aeruginosa isolates may share similar meropenem MICs, the differing resistance mechanisms produce discordant responses to a high-dose, prolonged infusion of meropenem. Thus, predicting the efficacy of an antimicrobial regimen based on MIC may not be a valid assumption for KPC-producing organisms.

Development of an HPLC Method for the Determination of Meropenem/Vaborbactam in Biological and Aqueous Matrixes.

A HPLC method was developed and validated to analyze meropenem and vaborbactam simultaneously in murine plasma and saline matrixes. A 60-µL volume of extracted sample was injected onto a 5-µm BDS Phenyl-Hypersil C18 reversed-phase column and analyzed with a UV detector set at 298 nm for the first 4.9 min and switched to 240 nm. The mobile phase contained a mixture of methanol and 25-mM sodium phosphate buffer set at a flow rate of 1.0 mL/min for the 16 min run time. Cefuroxime was used as the internal standard. The standard curves were linear over a range of 0.25-50 µg/mL. The precision and accuracy for 0.25 µg/mL (LLQ) in plasma for both compounds were <4.8% and >98.9%, respectively. Interday and intraday precision and accuracy for all QC plasma samples for both compounds were <6.2% and >95.7%, respectively. This methodology details a reproducible assay for both compounds using a single extraction with good accuracy and precision.

Hippocampal coding of conspecific position.

Many previous studies have shown that hippocampal place cells respond to the spatial position of the animal itself. Several recent studies have shown that place cells in an observer animal can also encode the location of a conspecific. The interpretation of these previous studies is, however, compromised by the fact that the observer animal was required to complete a movement that was either a duplication of the others trajectory, or a modification of it. This raises the possibility that the observed representation of the other, may have instead been a plan for the self. To test for a representation of a conspecific in a task where immediate behaviour was not immediately required of the observer, Sprague-Dawley rats were trained to run the length of a shuttle box for a food reward. They then observed a second animal (the runner) performing the same task. Positional data was obtained from the runner, while hippocampal single unit data was collected from the observer. Hippocampal single units were observed to have only limited, low resolution, firing rate-modulated representations of the runner animal. There was also evidence of a weak relationship between place cell spatial firing representations of the self and other. Some above-chance evidence of phase-coding of the runner's position was also observed in the observer animals, with an observer-centred reference frame. These results indicate that hippocampal place cells encode some limited spatial information about others when the observer's subsequent behaviour is not dependent on that of the observed.

Single ß-lactams versus combinations as empiric therapy for infections with Pseudomonas aeruginosa: assessing the in vitro susceptibility.

Background: While the value of combination versus monotherapy of infections with Pseudomonas aeruginosa infection is a subject of debate, increasing antimicrobial resistance of this pathogen makes it difficult to select appropriate empiric regimens. We evaluated the probability that P. aeruginosa would be susceptible to ß-lactams either as monotherapy or as part of a combination regimen.Methods: Contemporary non-duplicate isolates of P. aeruginosa derived from blood or the respiratory tract of patients hospitalized in the United States were investigated. Minimum inhibitory concentrations were determined using broth microdilution methods for amikacin (AMK), cefepime (FEP), ceftazidime (CAZ), ceftolozane/tazobactam (C/T), ciprofloxacin (CIP), fosfomycin (FOF), meropenem (MEM), piperacillin/tazobactam (TZP) and tobramycin (TOB). Susceptibility to a regimen was derived from the minimum inhibitory concentrations value of the beta-lactam plus the minimum inhibitory concentrations value of the additional agent.Results: In 1209 P. aeruginosa, susceptibility to C/T exceeded 90%, while susceptibility to FEP, CAZ, MEM and TZP ranged from 73 to 78%. For antibiotic combinations, the addition of the 2nd agent AMK, TOB, CIP or FOF raised the susceptibility to FEP, CAZ, MEM and TZP, whereas very little added activity was noted for C/T due to the intrinsic potency of this compound alone.Conclusions: While the addition of AMK, TOB, CIP or FOF markedly increased the probability that an active regimen would be selected for empirical therapy with FEP, CAZ, MEM and TZP, C/T alone had higher activity than the combinations.

Elevated vancomycin minimum inhibitory concentrations among methicillin-resistant Staphylococcus aureus isolated from patients with ventilator-associated pneumonia at a Connecticut hospital.

Emerging evidence suggests that vancomycin minimum inhibitory concentrations (MICs) within methicillin-resistant Staphylococcus aureus (MRSA) are increasing. The objective of this surveillance study was to determine vancomycin MIC distributions for MRSA isolates collected from the respiratory tract of patients with ventilator-associated pneumonia (VAP) at a large community hospital in Hartford, Connecticut. The frequency of heteroresistant vancomycin intermediate S. aureus (hVISA) was also assessed for select isolates. Vancomycin MICs and hVISA screening were conducted using standard inoculum and macromethod Etest methodology, respectively, for isolates collected between November 2005 and August 2007. Fifty-eight isolates of MRSA were collected over the two-year period. The MIC50 and MIC90 were both 2 microg/ml; 31.0% and 58.6% of isolates had vancomycin MICs of 1.5 microg/ml and 2 microg/ml, respectively. None of the isolates tested were positive for hVISA; four isolates were VISA. Vancomycin MICs for respiratory MRSA at this Connecticut hospital are elevated. Institution-specific surveillance in the state is warranted.

Development of an HPLC-MS/MS method for the determination of ceftolozane/tazobactam in bronchoalveolar lavage fluid.

AIM: We describe the validation of an HPLC-MS/MS method to analyze ceftolozane and tazobactam simultaneously in saline matrixes. MATERIALS & METHODS: An Agilent 1260 HPLC interfaced to an Agilent 6470 triple-quadrupole mass spectrometer was used for quantification. A reverse-phase column running a gradient of water and acetonitrile containing 0.1% formic acid mobile phase at a flow rate of 1.0 ml/min provided chromatographic fractionation. Tazobactam15N3 was used as the internal standard. The standard curves were linear over a range of 0.02-0.5 µg/ml. CONCLUSION: This methodology represents a simple, reproducible approach to the determination of drug concentrations with accuracy and precision for pharmacokinetic studies undertaken with this recently US FDA-approved antimicrobial therapy.