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A prospective observational study was performed to determine whether recurrent episodes of pneumonia caused by Pseudomonas aeruginosa in ventilated patients were due to a relapse of the previous clone or to reinfection with a new one. Diagnosis was based on quantitative cultures of secretions obtained by bronchoscopy. Comparison of strains was made by chromosomal fingerprinting based on pulsed field gel electrophoresis (PFGE). Thirty-three (89.1%) of 37 patients survived the initial week after pneumonia diagnosis; six survivors (18.1%) had multiple episodes caused by the same species. Presence of adult respiratory distress syndrome (83.3% versus 22.2%, p = 0.02) was the only factor significantly associated with clinical recurrences. The 16 isolates from five patients (nine recurrences) were analyzed by PFGE. All new isolates from recurrent episodes, excepting one, were considered as relapses. These data suggest that most recurrent episodes of P. aeruginosa pneumonia in ventilated patients occur due to persistence of strains present in a prior infection.

PurposeClarithromycin was considered the cornerstone for the treatment of Mycobacterium abscessus complex infections. Genetic resistance mechanisms have been described and many experts propose amikacin as an alternative. Nevertheless, clarithromycin has several advantages; therefore, it is necessary to identify the non-functional erm(41) allele to determine the most suitable treatment. The aims of this study were to characterize the molecular mechanisms of clarithromycin resistance in a collection of Mycobacterium abscessus complex isolates and to verify the relationship between these mechanisms and the antibiogram.Materials and MethodsClinical isolates of M. abscessus complex (n = 22) from 16 patients were identified using four housekeeping genes (rpoB, secA1, sodA and hsp65), and their genetic resistance was characterized by studying erm(41) and rrl genes. Nine strains were recovered from the clinical isolates and subjected to E-test and microdilution clarithromycin susceptibility tests, with readings at 3, 7 and 14 days.ResultsWe classified 11/16 (68.8%) M. abscessus subsp. abscessus, 4/16 (25.0%) M. abscessus subsp. bolletii, and 1/16 (6.3%) M. abscessus subsp. massiliense. T28 erm(41) allele was observed in 8 Mycobacterium abscessus subps. abscessus and 3 Mycobacterium abscessus subsp. bolletii. One strain of M. abscessus subsp. bolletii had an erm(41) gene truncated and was susceptible to clarithromycin. No mutations were observed in rrl gene first isolates. In three patients, follow-up of initial rrl wild-type strains showed acquired resistance.ConclusionsMost clinical isolates of M. abscessus complex had inducible resistance to clarithromycin and total absence of constitutive resistance. Our findings showed that the acquisition of resistance mutations in rrl gene was associated with functional and non-functional erm(41) gene. Caution is needed when using erm(41) sequencing alone to identify M. abscessus subspecies. This study reports an acquired mutation at position 2057 of rrl gene, conferring medium-low clarithromycin constitutive resistance.

Highlights d NHEJ-mediated gene editing enables highly efficient editing in human long-term HSCs d NHEJ-mediated editing restores mutant coding frames across FA complementation groups d Corrected FA-HSCs have a marked proliferative advantage in vitro and in vivo