When Vif was co-expressed with CBFb140-His, the solubility of Vif improved significantly

y period. 2: Neurophysiological assessments in the peripheral nerves. Recordings of the NCV and neuronal action potential amplitude were performed on the digital and caudal nerves as a measure of the sensory and sensory/motor neurophysiological functionality, Cobicistat site respectively. As reported in previous studies Carozzi et al., 2010a), treatment with bortezomib induced a reduction of the caudal NCV and amplitude and of the digital NCV, but not of the digital amplitude. 3: Assessments of mechanical and thermal thresholds. The mice were tested at baseline, i.e. before starting drug treatment, and then weekly for 4 weeks to assess the nocifensive response to mechanical and thermal stimuli. The development of mechanical allodynia was determined using the Dynamic Aesthesiometer Test and was defined as a decrease in the paw withdrawal threshold compared to the vehicle and naive groups. No significant difference in withdrawal threshold was found between the groups at baseline. However, the bortezomib-treated mice had a significant decrease in mechanical threshold compared to the nave and vehicle-treated group starting after 1 week of drug treatment and persisting through the fourth week. 10336542 There was no statistically significant difference 2173565 in the hot or cold response thresholds between the groups of mice at any time point after bortezomib treatment. 4: Assessments of spinal cord neuronal electrical activity. After determining that bortezomib induced the development of painful PN characterized by impaired neurophysiological function of peripheral nerves and by altered nocifensive behavioral responses to mechanical stimuli, we investigated whether bortezomib treatment also induced changes in the activity of WDR neurons in the spinal cord dorsal horn. The electrical activity of 68 deep WDR neurons in the lumbar enlargement of the spinal dorsal horn was measured. Extracellular electrophysiological recording demonstrated that the number of spikes per second was significantly higher in bortezomib-treated mice compared to 6 Experimental Bortezomib Peripheral Neuropathy doi: 10.1371/journal.pone.0072995.g003 nave and vehicle-treated mice in response to innocuous, moderate and noxious stimulation of the hind paw ipsilateral to the spinal recording site. 5: Assessment of different sensory fiber function. Sensory fiber function was assessed by determining the CPT in the hind paw using the Neurometer device, which delivers transcutaneous electrical stimuli at frequencies that have been previously reported to be specific to each fiber type. Bortezomib treatment induced a significant and persistent decrease in the amount of current required to induce a paw withdrawal to the 5 Hz and 250 Hz stimulation frequencies. By contrast, there was only a small difference in the amount of current required to elicit a paw withdrawal using the 2000 Hz stimulation between the drug and vehicle-treated groups, which resolved back to baseline in the third week. These data suggest that the A and C fibers are sensitized early after bortezomib treatment in hind paw, such that less current is required to stimulate a response. 6: Assessment of morphological alterations: light microscopy. Morphological examination of the peripheral nerves, dorsal/ventral roots, DRG and lumbar spinal cord was done to determine whether any pathological changes were present. Three mice from each group were sacrificed after the last administration of bortezomib and used for the sample collection and processing.