Kohler, P. | Search bioRxiv

bioRxiv2026

pH-Dependent Evolution of Delafloxacin and Ciprofloxacin Resistance in Pseudomonas aeruginosa from cystic fibrosis (CF) and non-CF-patients

Delafloxacin (DLX) is a novel fluoroquinolone with enhanced antibacterial activity in acidic environments, a property that may be advantageous for treating Pseudomonas aeruginosa infections in cystic fibrosis (CF), where airway surface liquid pH is typically reduced (pH 5.5-6.7). However, the propensity for resistance development and the underlying mechanisms in P. aeruginosa remain incompletely defined. We conducted serial passage experiments on six clinical P. aeruginosa isolates (three CF-derived and three non-CF-derived) exposed to sub-inhibitory concentrations of DLX or ciprofloxacin (CIP) at pH 6.0 and 7.3 over nine days. Susceptibility was assessed by broth microdilution (BMD), and resistance mechanisms were characterized by whole-genome sequencing (WGS), efflux pump expression analysis (qRT-PCR), and functional validation using CRISPR/Cas9-mediated genome editing and complementation assays. DLX minimal inhibitory concentrations (MICs) rose only 10.1- to 28.5-fold over 9 days, compared with 77.6- to 97.8-fold for CIP, indicating a substantially higher genetic barrier to resistance. This barrier was most pronounced under acidic conditions: only 38.9% of DLX-passaged samples crossed the resistance breakpoint, compared with 94.4% at neutral pH, whereas CIP resistance reached 100% regardless of pH. Cross-resistance was asymmetric: exposure to DLX consistently selected for CIP cross-resistance (97.2% of samples), whereas exposure to CIP induced DLX cross-resistance efficiently at neutral pH but only partially under acidic conditions. A previously undescribed gyrA mutation (p.Ala51Val) conferred a 4-fold increase in DLX MIC when introduced by CRISPR/Cas9, and upregulation of the MexEF-OprN efflux pump, reversible by mexS complementation, emerged as a prominent resistance mechanism. Overall, DLX exhibited a markedly higher genetic barrier to resistance than CIP in P. aeruginosa, particularly under the acidic conditions characteristic of the CF airway. However, its use may co-select for CIP cross-resistance through efflux upregulation, underscoring the need for careful stewardship in CF.

microbiology↗

bioRxiv2025

Reorganization of motor functions within visuomotor networks subsequent to somatosensory cortical damage

Somatosensory inputs are critical to motor control. Animal studies have shown that primary somatosensory lesions cause sensorimotor deficits along with disrupted organization in primary motor cortex (M1). How does damage in primary somatosensory cortex (S1) influence motor networks in humans? Using fMRI, we examined two individuals with extensive damage to left somatosensory cortex, but primarily intact motor cortex and preserved motor abilities. Given left S1 damage, tactile detection and localization were impaired for the contralesional hand in both individuals. When moving the contralesional hand, LS, with near complete damage to the S1 hand area, showed increased activation in ipsilesional putamen and deactivation in contralesional cerebellum relative to age-matched controls. These findings demonstrate influences of S1 damage to subcortical sensorimotor areas that are distant from the lesion site, and a potential reweighting of the motor network with increased action selection in putamen and inhibition of sensory prediction in cerebellum in the face of sensory loss. In contrast, RF, who had a small island of spared S1 in the hand area, showed greater activation in contralesional S1 for movement versus rest. This same region was also activated by pure somatosensory stimulation in a second experiment, suggesting that the spared S1 area in RF still subserves sensorimotor processing. Finally, the right middle occipital gyrus was more strongly activated in both individuals compared with controls, suggesting a potential reliance on visual imagery in the face of degraded sensory feedback.

neuroscience↗