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Abstract High-pressure liquid chromatographic (HPLC) methods were developed for the analysis of ciprofloxacin hydrochloride raw materials. Method A is for drug content and the determination of related compounds eluting before the drug, including the ethylenediamine analog of ciprofloxacin. Method B...
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PCR was used to amplify a 238-bp region from Helicobacter pylori which corresponded to the quinolone resistance-determining region in Escherichia coli. The gyrA gene of H. pylori was cloned and sequenced. An open reading frame of 2,478 nucleotides coded for a polypeptide of 826 amino acids with a calculated molecular mass of 92,508 Da. The amino acid sequence showed an overall 52% identity with other bacterial gyrA genes but was most closely related to the gyrA subunit of Campylobacter jejuni (76.5% identity). Sequencing of the amplification product from ciprofloxacin-resistant mutants of H. pylori revealed four classes of mutations with substitutions at amino acid 87 (Asn--,Lys), amino acid 88 (Ala--,Val), and amino acid 91 (Asp--,Gly, --,Asn, or --,Tyr) and a double substitution at amino acids 91 and 97 (Ala--,Val). Ciprofloxacin-susceptible strains of H. pylori could be transformed to ciprofloxacin resistance by using the amplified fragment from resistant strains as donor DNA. Of the 11 ...
High-level ciprofloxacin resistance (MIC ≥ 64 μg/ml) in E. faecium is associated with amino acid changes in topoisomerase IV and DNA gyrase and confined to a single clonal complex, previously identified as a global hospital-adapted subpopulation of E. faecium (CC17). CC17 is further characterized by ampicillin resistance and a pathogenicity island, including the variant esp gene (20, 31, 32, 65) (Fig. 1). Cipr isolates belonging to other genetic backgrounds are only low-level resistant (MIC = 4 μg/ml) and do not carry these point mutations. These findings further disclose the cumulative adaptive mechanisms that have resulted in the evolution of a hospital-adapted genetic complex in E. faecium.. We have previously described the global spread of CC17 E. faecium, which is characterized by ampicillin resistance, although pandemic spread of VRE was instrumental for its identification. The strong association with quinolone resistance strongly suggests that this clonal complex has previously ...
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The flow rate is about 1.5 mL per minute. Chromatograph the Resolution solution, and record the peak responses as directed for Procedure: the retention time for ciprofloxacin is between 6.4 and 10.8 minutes; the relative retention times are about 0.7 for ciprofloxacin ethylenediamine analog and 1.0 for ciprofloxacin; and the resolution, R, between the ciprofloxacin ethylenediamine analog peak and the ciprofloxacin peak is not less than 6. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the column efficiency, determined from the ciprofloxacin peak, is not less than 2500 theoretical plates; the tailing factor for the ciprofloxacin peak is not more than 2.0; and the relative standard deviation for replicate injections is not more than 1.5 ...