After 6 months from the last stimulation we selected all patients who had achieved and maintained a benefit from the treatment (n=12).
One to six days following the last stimulation session, forelimb movement representations were derived using high-resolution intracortical microstimulation in the left sensorimotor neocortex.
Animals were sacrificed 24 h after last stimulation and their brain used for labeling μ opioid, benzodiazepine (BZD), α1-adrenergic, and adenylyl cyclase binding.
Three months after the last stimulation, headache frequency, headache days, duration and intensity (VAS), per month were evaluated for each patient: 80% of the patients (n=9) obtained a further reduction of at least 50% of the headache parameters evaluated.
The experimenter validated the program's interpretation (i.e 'NFR detected' or 'no NFR') using an online graphic display of the EMG from the timeframe surrounding the last stimulation.
Data normalization was performed by calculating the ratio of a given response to the average response elicited by the ligand alone (first and last stimulation) (Fig. S6).
Six EEG intervals were selected for the analyses: a 1-min period immediately before theta-tDCS (baseline) and the five 1-min stimulation-free periods immediately after each 5-min block of stimulation (including the interval after the last stimulation).
Seven days after the last stimulation with CD40L and IL-4, B cells were harvested, washed twice, distributed to 48-well plates (0,25 million/0,5 ml/well) on top of a layer of irradiated CD40L-expressing or non-expressing murine L fibroblasts (20,000/well), and stimulated with phosphorothioate oligonucleotides at 4 µg/ml for 4 days.
After the last stimulation with EA, the rats were sacrificed.
After the last stimulation with EA, all rats were sacrificed.
