Fluorimetric Detection of a Bacillus stearothermophilus Spore-Bound Enzyme, alpha-d-Glucosidase, for Rapid Indication of Flash Sterilization Failure.

A biological indicator based on fluorimetric detection within 60 min of a Bacillus stearothermophilus spore-bound enzyme, alpha-d-glucosidase, has been developed. Results indicate that the enzyme survived slightly longer than spores observed after 24 h of incubation. The new system shows promise for evaluating flash sterilization cycles within 60 min compared with conventional 24-h systems.

Synthesis, absolute configuration, and antibacterial activity of 6,7-dihydro-5,8-dimethyl-9-fluoro-1-oxo-1H,5H- benzo[ij]quinolizine-2-carboxylic acid.

The tricyclic quinolone antibacterial agent 6,7-dihydro-5,8-dimethyl-9-fluoro-1-oxo-1H,5H-benzo[ij]quinolizine -2-carboxylic acid has an asymmetric center at position 5 of the molecule. The R and S isomers of the compound have been prepared from the corresponding (R)- and (S)-2,5-dimethyl-6-fluoro-1,2,3,4-tetrahydroquinolines, which were separated via their diastereomeric amides of N-tosyl-(S)-proline. The absolute configuration was established by X-ray analysis of one of the diastereomeric amides. The 5-desmethyl analogue was prepared for antibacterial comparison with the isomers and the racemic mixture. It has now been established that the S isomer is much more active than the R isomer. The 5-desmethyl analogue was found to be more active than the R isomer but not as active as the S isomer or the racemic mixture. The importance of stereochemistry at position 5 in this system has been established.

Disposition of radiolabeled flumequine in rat and dog.

Single doses of 14C-flumequine (a urinary tract antibacterial) were given to rats and dogs. Unchanged drug accounted for more than 80% of the drug in plasma of rats but only 25-50% in plasma of dogs. Although only a small percentage of the dose was excreted in the urine of each species as unchanged drug, a wide differentiation in the urinary metabolite profiles was observed. No significant tissue retention was seen in rats or dogs.

Differentiation of fluorinated quinolone antibacterials with Neisseria gonorrhoeae isolates.

Clinical isolates of Neisseria gonorrhoeae were tested for growth inhibition by various quinolones. One acrosoxacin-resistant isolate was also resistant to several fluorinated quinolones with 7-heterocyclic substitution, but, inhibitory concentrations were not elevated for quinolones substituted with a methyl group in the corresponding position (nalidixic acid:position 7, S-25930:position 8) or unsubstituted (flumequine:position 8). Isolates resistant to flumequine demonstrated elevated inhibitory concentrations for all quinolones examined.

Plasma and urine levels of flumequine and 7-hydroxyflumequine following single and multiple oral dosing.

Plasma and urine concentrations of flumequine and its microbiologically active metabolite, 7-hydroxyflumequine, were determined in healthy subjects following single oral doses of 400, 800, and 1200 mg of flumequine, and following multiple oral doses of 800 mg given four-times daily. After administration of the single oral doses, antimicrobial levels in plasma and urine were rapidly attained, were proportional to the dose given, and were maintained for 12 to 24 h. The multiple dosage regimen yielded antimicrobial levels in both plasma and urine that were several-fold higher than the levels required to inhibit the growth of susceptible bacteria. Following both the single and multiple dose regimens, the plasma elimination half-life of flumequine was about 7h. The excretion of 7-hydroxyflumequine in the urine contributed significantly to the antimicrobial activity.

Determination of flumequine and a hydroxy metabolite in biological fluids by high-pressure liquid chromatographic, fluorometric, and microbiological methods.

A sensitive and specific high-pressure liquid chromatographic method is described for the determination of the antibacterial drug flumequine and a major metabolite, 7-hydroxyflumequine, in human plasma and urine. The assay was linear over a concentration range of 1 to 120 micrograms/ml for both compounds. This method is compared with fluorometric and microbiological assays for flumequine. These latter methods did not differentiate between flumequine and any fluorescent or antimicrobiologically active metabolites. However, because essentially all drug in the plasma was found to be flumequine in radiolabeled studies, levels of unchanged drug in the plasma could be quantitated by either high-pressure liquid chromatography or fluorometry. Although only high-pressure liquid chromatography was able to specifically measure flumequine in the urine, the antimicrobial activity of the urine, which is more therapeutically relevant due to antimicrobially active metabolites, could be quantitated by either the fluorometric or the microbiological assay.

Bioevaluation of the antibacterial flumequine for urinary tract use.

The antimicrobial activity of flumequine (R-802) was characterized by in vitro and in vivo procedures. Assay of the minimal inhibitory concentrations for 321 recent clinical isolates revealed that 88% of the gram-negative bacteria were inhibited by an R-802 concentration of 6.2 mug/ml or less. Cross-resistance in laboratory-derived mutants of Proteus vulgaris was essentially complete for R-802, nalidixic acid, and oxolinic acid, although quantitative differences were evident. R-802 was more effective than either of these quinolone antibacterials in preventing the development of experimental murine pyelonephritis (P. vulgaris). R-802 and trimethoprim/sulfamethoxazole (1:5) were equally effective in resolving a P. mirabilis-induced prostatitis of rats.

Purification and characterization of the beta-galactosidase of Aeromonas formicans.

Rohlfing, S. R. (Western Reserve University, Cleveland, Ohio), and I. P. Crawford. Purification and characterization of the beta-galactosidase of Aeromonas formicans. J. Bacteriol. 91:1085-1097. 1966.-The beta-galactosidase of Aeromonas formicans was purified by diethylaminoethyl cellulose chromatography and gel filtration on Sephadex G-200. The properties of the enzyme molecule were compared with purified beta-galactosidase from Escherichia coli. The sedimentation coefficients and electrophoretic mobilities of the two enzymes were not significantly different; the electrophoretic mobility of urea-produced subunits of the two enzymes was also similar. The stabilities of the two enzymes to denaturing agents provided measurable differences; E. coli beta-galactosidase is relatively more heat-stable and more resistant to the action of urea. The amino acid compositions of the two proteins revealed significant differences in several amino acids, particularly alanine, arginine, glycine, and leucine. The comparisons cited suggest that A. formicans and E. coli are not completely unrelated, for their beta-galactosidases show considerable structural similarity.