Some UMI-77 Common Myths Uncovered
GyrA (Ile83Ser, numbering according to E.?coli) and ParC (Ser80Ile) modifications were identified in all the isolates supporting the significant quinolone MICs in addition to PA��N-sensitive efflux. The signals obtained from RT-PCR analysis did not systematically correlate with the functional assembly of bacterial membrane proteins [13]. Consequently, the analyses were carried out by staining the membrane protein fractions and by performing western blot analysis [13,14]. For IMI-S isolates, as indicated in Table?2 and Fig.?1 for the strains of patient E, the AcrA and TolC signal observed was slightly higher than that of the reference strain ATCC 13048. Outer membrane proteins OmpA and OmpX were detected at the same expression level (Table?2). For Omp35 and Omp36, all IMI-S isolates exhibited a positive signal (Table?2). For IMI-I/R strains, an increased level of AcrA and TolC, similar to that of strain EA27, was observed in UMI-77 clinical trial all strains except for strain NEA736500SCR (Patient E). This isolate exhibited the highest signal for TolC (Fig.?1 and Table?2). No change of OmpA and OmpX expression level was detected (Table?2). For the Omp35 signal three profiles can be observed: (i) an Omp35 expression in IMI-I strains for which IMI MICs were 4�C8?mg/L (n?=?4); (ii) a decreased Omp35 level and the emergence of a new band for NEA736500BCR for strains having an IMI MIC 16?mg/L (n?=?1); and (iii) no expression of Omp35 in IMI-R strains, IMI MICs between 16�C32?mg/L (n?=?3). Similar results were obtained with antibodies directed against Omp36 (Table?2). The new band, observed in isolate NEA736500BCR, was CYTH4 identified by mass spectrometry to an OmpA-like protein. The HPI virulence factor-encoding genes were detected in all isolates. The C.?elegans model is well described for the study of bacterial factors involved in infection and colonization [26,27]. In the nematode killing assay, all IMI-S E.?aerogenes isolates were significantly more virulent (LT50 3.5�C4.3?��?0.5?days; LT100 7.8�C8.3?��?0.9?days) than both IMI-I and IMI-R isolates (LT50 5.1�C6.9?��?0.8?days; LT100 10.3�C13.5?��?0.7?days) (p?click here respectively) (p?
