The Natural Product Elegaphenone Potentiates Antibiotic Effects against Pseudomonas aeruginosa.

Natural products represent a rich source of antibiotics that address versatile cellular targets. The deconvolution of their targets via chemical proteomics is often challenged by the introduction of large photocrosslinkers. Here we applied elegaphenone, a largely uncharacterized natural product antibiotic bearing a native benzophenone core scaffold, for affinity-based protein profiling (AfBPP) in Gram-positive and Gram-negative bacteria. This study utilizes the alkynylated natural product scaffold as a probe to uncover intriguing biological interactions with the transcriptional regulator AlgP. Furthermore, proteome profiling of a Pseudomonas aeruginosa AlgP transposon mutant provided unique insights into the mode of action. Elegaphenone enhanced the elimination of intracellular P. aeruginosa in macrophages exposed to sub-inhibitory concentrations of the fluoroquinolone antibiotic norfloxacin.

Killing by bactericidal antibiotics does not depend on reactive oxygen species.

Bactericidal antibiotics kill by modulating their respective targets. This traditional view has been challenged by studies that propose an alternative, unified mechanism of killing, whereby toxic reactive oxygen species (ROS) are produced in the presence of antibiotics. We found no correlation between an individual cell's probability of survival in the presence of antibiotic and its level of ROS. An ROS quencher, thiourea, protected cells from antibiotics present at low concentrations, but the effect was observed under anaerobic conditions as well. There was essentially no difference in survival of bacteria treated with various antibiotics under aerobic or anaerobic conditions. This suggests that ROS do not play a role in killing of bacterial pathogens by antibiotics.

Distribution of gyrase and topoisomerase IV on bacterial nucleoid: implications for nucleoid organization.

We explored the existence of nucleoid DNA loops in Escherichia coli by studying the distribution of bacterial type II topoisomerases (Topo IIs). Norfloxacin-induced high molecular weight (HMW) DNA fragmentation of nucleoid, an event reminiscent of the excision of eukaryotic chromosomal DNA loops mediated by topoisomerase II (TOP2). The size of the HMW DNA fragments induced by norfloxacin was affected by transcription, translation and growth phases of bacteria. The involvement of bacterial Topo IIs in the generation of these HMW DNA fragments is supported by the following observations: (i) the excised loop-sized DNA fragments were covalently linked to proteins; (ii) the norfloxacin-induced excision of DNA loops was highly reversible; (iii) coumermycin A1 antagonized the excision of DNA loops induced by norfloxacin; (iv) this antagonistic effect was reduced in either gyrase or topo IV mutants conferring coumarin resistance and (v) norfloxacin-induced reversible, gyrase-mediated DNA cleavage in vitro. Importantly, studies on coumarin- and/or quinolone-resistant mutant strains showed that DNA gyrase, rather than topoisomerase IV, plays the major role in the generation of loop-sized HMW DNA fragments. In sum, our study suggests a potential role of Topo IIs in the arrangement of DNA supercoiling loop domains in prokaryotic cells.

[Synthesis and antibacterial activity of pyridonecarboxylic acid derivatives containing 2-methyl-5-nitroimidazol].

AIM: To study the synthesis and antibacterial activity of pyridonecarboxylic acid derivatives containing 2-methyl-5-nitroimidazol. METHODS: Pyridonecarboxylic acid derivatives containing 2-methyl-5-nitroimidazol were synthesized primarily from 2-methyl-5-nitroimidazol, norfloxacin, ciprofloxacin, enoxacin via nucleophilic substitution and esterification. The antibacterial activity of the nine target compounds were tested. RESULTS: Nine new compounds were synthesized (IIa-c and IIIa-f). The structure of the title compounds were identified by 1HNMR, MS as well as elementary analysis. CONCLUSION: Compounds IIa, IIb and IIc showed antibacterial activity, and were worth further studying.

[Yersinia pseudotuberculosis sensitivity to fluoroquinolones and the efficacy of pefloxacin in experimental pseudotuberculosis]. / Izuchenie chuvstvitel'nosti Yersinia pseudotuberculosis k ftorkhinolonam i éffektivnosti pefloksatsina pri éksperimental'nom psevdotuberkuleze.

The sensitivity of Y. pseudotuberculosis of different origin and plasmid profile to Russian preparations of the ftoroquinolones group (cyprofloxacin, pefloxacin, norfloxacin) was studied. Of the 3 preparations under study, pefloxacin was found to be most active with respect to Y. pseudotuberculosis. The absence of relationship between antibiotic resistance in 4 Y. pseudotuberculosis strains to cyprofloxacin, pefloxacin and norfloxacin and their additional conjugative R-plasmid with molecular weights of 32 and 38 mD was noted. Pefloxacin was shown to give a good sanitation and protective effect in experimental pseudotuberculosis.

Antagonism between bactericidal activities of 4-quinolones and coumarins gives insight into 4-quinolone killing mechanisms.

At concentrations exceeding their MICs, novobiocin and coumermycin antagonised the bactericidal activities of nalidixic acid, ciprofloxacin, ofloxacin and norfloxacin against Escherichia coli KL16. The sensitivities to killing by ciprofloxacin of four mutant derivatives of KL16 carrying gyrA, nalB, nal24 or nal31 alleles were also antagonised by novobiocin. The activities of drug combinations were tested in nutrient broth, which allowed expression of 4-quinolone killing mechanisms A, B and C. They were also tested in nutrient broth plus rifampicin to inhibit mechanisms A and C of the 4-quinolones, and in phosphate-buffered saline, which inhibited mechanism A. Results showed that novobiocin antagonised mechanism C, but not B, of both ciprofloxacin and ofloxacin, but did not antagonise mechanism C of norfloxacin. A review of these and other data indicates that mechanism B may result from the activities of SOS error-prone DNA repair on an irreversibly-bound drug-gyrase-DNA complex, and that mechanism C is mediated via drug interaction with the B subunit of DNA gyrase.

Effects of fluoroquinolone antimicrobials alone and in conjunction with theophylline on seizures in amygdaloid kindled rats. Mechanistic and pharmacokinetic study.

The influence of three fluoroquinolone (FQ) antimicrobial drugs (ciprofloxacin (CP), norfloxacin (NF), enrofloxacin (EF)) on seizure parameters in amygdaloid kindled rats was investigated. CP and NF (100 mg/kg i.p.) did not modify seizure parameters while EF induced a decrease in seizure activity. Since clinical data indicate a seizure enhancing interaction between FQ and theophylline (THEO) we studied the influence of concurrent FQ-THEO administration in kindled rats. CP and NF, but not EF given concurrently with a non-seizure modulating dose of THEO (10 mg/kg i.p.) caused increases in seizure activity and aggressiveness in the animals. The CP-THEO induced seizure enhancement was antagonized by 2-chloroadenosine and diazepam. Pharmacokinetic studies demonstrated that THEO serum levels and elimination were not altered by concurrent CP administration. We conclude that coadministration of FQ-THEO can aggravate amygdala kindled seizures and that this aggravation may involve centrally mediated mechanisms.

Activity and mechanism of action of DuP 105 and DuP 721, new oxazolidinone compounds.

The in-vitro activities of DuP 721 and DuP 105, new oxazolidinone antibacterials, were compared with those of cefazolin, cephalexin, ciprofloxacin, clindamycin, oxacillin, penicillin, and vancomycin against Gram-positive cocci. DuP 721 was approximately four-fold more active than DuP 105 with an MIC of 2.0 mg/l for 90% of the Staphylococcus aureus, beta-haemolytic streptococcus and Streptococcus faecalis strains tested, and an MIC of 4.0 mg/l for 90% of the Str. faecium, penicillin-resistant Str. pneumoniae and viridans streptococcus strains tested. DuP 105 was most active against strains of Staph. epidermidis with an MIC of 4.0 mg/l for 90% of the strains tested. There was no cross resistance between these and the other antibacterial agents that were tested. Both oxazolidinones had bacteriostatic activity in broth against susceptible organisms. Both DuP 721 and DuP 105 inhibited ribosomal protein synthesis in a cell-free system. These synthetic, orally absorbable compounds represent a new series of antibacterial agents unrelated by chemical structure to any other currently available antimicrobial agents.