Assessment of urinary pharmacokinetics and pharmacodynamics of orbifloxacin in healthy dogs with ex vivo modelling.

PURPOSE: The aim of this study was to investigate the urinary pharmacokinetics (PK) of orbifloxacin (OBFX) administered at 5 mg kg-1 in six healthy dogs. A further aim was to use an ex vivo model to evaluate the urinary PK and pharmacodynamics (PD) of OBFX to determine its urinary bactericidal titre (UBT), which represents the maximal dilution of urine allowing bactericidal activity. METHODOLOGY: Fourteen urinary tract infection (UTI) pathogenic strains of five bacterial species (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis and Staphylococcuspseudintermedius) were used. Urine samples were obtained every 4 h for the first 24 h after OBFX administration, for measurement of urine drug concentration and UBT.Results/Key findings. The urine OBFX concentration peaked at 0-4, 4-8 or 4-8 h after administration, with a maximum concentration of 383±171 µg ml-1. Overall, the fluctuation in median UBT closely correlated with that of the mean urine OBFX concentration. In addition, the median areas under the UBT-time curves (AUBTs) were significantly inversely correlated with the MICs for OBFX in the tested strains (P<0.01). Notably, median UBTs and AUBTs were extremely low (0-0.5 and 2-5, respectively) in OBFX-resistant E. coli strains with MIC ≥8 µg ml-1. CONCLUSION: The fluctuation of UBTs closely correlated with that of urine concentration, and UBT values depended on the susceptibility of the bacterial strains to OBFX. We believe that ex vivo modelling to determine UBTs is useful to evaluate the urinary PK/PD of antimicrobials indicated for UTIs in dogs.

Are liquid chromatography/electrospray tandem quadrupole fragmentation ratios unequivocal confirmation criteria?

Multiple reaction monitoring (MRM) ratios as provided by tandem mass spectrometers are used to confirm positive residue findings (e.g. veterinary drugs or pesticides). The Commission Decision 2002/657/EEC defines tolerance levels for MRM ratios, which are intended to prevent the reporting of false positives. This paper reports findings where blank sample extracts have been spiked by a drug (difloxacin) and the corresponding measured MRM ratios significantly deviated from MRM ratios observed in matrix-free solution. The observation was explained by the formation of two different [M+H](+) analyte ions within the electrospray ionization (ESI) interface. These two ions vary only by the site of analyte protonation. Since they are isobaric, they are equally transmitted through the first quadrupole, but are differently fragmented in the collision chamber. The existence of two isobaric ions was deduced by statistical data and the observation of a doubly charged analyte ion. It was hypothesized that the combined presence of [M+H](+) and [M+2H](2+) implies the existence of two different singly charged ion species differing only by the site of protonation. Low- and high-energy interface-induced fragmentation was performed on the samples. The surviving precursor ion population was mass selected and again fragmented in the collision chamber. Equal product ion spectra would be expected. However, very different product ion spectra were observed for the two interface regimes. This is consistent with the assumption that the two postulated isobaric precursor ions show different stability in the interface. Hence the abundance ratio among the two types of surviving precursor ions will shift and change the resulting product ion spectra. The existence of the postulated singly charged ions with multiple chargeable sites was finally confirmed by successful ion mobility separation.

Use of scintigraphy for assessment of fracture healing and early diagnosis of osteomyelitis following fracture repair in rabbits.

OBJECTIVE: To evaluate use of technetium Tc 99m disodium hydroxymethylene diphosphonate (99m-Tc-HDP) for assessing fracture healing and 99m-Tc-HDP and technetium Tc 99m ciprofloxacin (99m-Tc-CIPRO) for early diagnosis of osteomyelitis in rabbits. ANIMALS: 32 skeletally mature New Zealand White rabbits. PROCEDURE: A femoral fracture defect stabilized with bone plates and cortical screws was used. Scintigraphy was performed 4, 8, 12, and 16 weeks after surgery. The 99m-Tc-CIPRO scan was performed 48 hours after the 99m-Tc-HDP scan. The uptake ratio of the experimental limb to the normal limb was calculated by use of multiple regions of interest. Results of radiography performed to determine external callus and lysis grade and percentage defect ossification at 16 weeks were compared with scintigraphy results. RESULTS: Infected fractures had a higher uptake ratio for 99m-Tc-HDP and 99m-Tc-CIPRO than noninfected fractures. Infected fractures could be differentiated from noninfected fractures late in healing by use of 99m-Tc-HDP. Although 99m-Tc-CIPRO was better than 99m-Tc-HDP for identifying infection, there was a high incidence of false positive and negative results with 99m-Tc-CIPRO. There was an association between 99m-Tc-HDP uptake ratio and callus formation and a good correlation between 99m-Tc-HDP uptake ratio and defect ossification after 4 weeks. CONCLUSIONS AND CLINICAL RELEVANCE: 99m-Tc-HDP and 99m-Tc-CIPRO may be useful for diagnosing osteomyelitis late in fracture healing; however, false positive and false negative results occur. Technetium Tc 99m disodium hydroxymethylene diphosphonate may be useful for evaluating fracture healing.

An assessment of cerebral blood flow and metabolism after fleroxacin therapy.

Cerebral blood flow (CBF), glucose (FDG), and oxygen metabolism (OM) were evaluated by positron emission tomography (PET) in 18 healthy volunteers who were randomized to a 72-hour course of either 600 mg/d of fleroxacin or placebo. Such studies attempted to assess potentially serious, yet unexplained, central nervous system adverse effects of the fluorinated quinolone class. Baseline and postplacebo values for CBF (mL/min/100 g) and FDG (mg/min/100 g) were: 53 +/- 6 and 5.7 +/- 1.8; and 49.6 +/- 4.4, and 5.2 +/- 1.2, respectively. Identical values for fleroxacin were: 53.9 +/- 4.8 and 6.3 +/- 1.1; and 54.4 +/- 2.2 and 6.8 +/- 1.5, respectively. The differences between fleroxacin and placebo were not significant. There was also no effect seen in OM between placebo and the active drug. The authors conclude that short-term administration of fleroxacin does not alter CBF, FDG, or OM in healthy volunteers.