In an effort to get a clearer signal for a potential dose response, we are making minor adjustments to the next series of experiments taking place April 22 and 23, 2021 with our second therapist.
Instead of delivering the treatments over 2 consecutive days for each dose, treatment will be delivered to each set of petri dish only once and we will test the following treatment doses: 5, 15, 30 and 60 minutes.
We will conduct two separate experiments each day for treatment group, sham control group, and normal controls that stay in the incubator. We will only test Panc-1 for these next two days of experiments.
**DAY ONE:**
- Experiment 1: Dose: 5 minutes; 30 minutes; 15 minutes
- Experiment 2: Dose: 15 minutes; 60 minutes; 30 minutes
Outcomes assessed in both experiments include: 2D Prestoblue proliferation assay assessed at 0 hour after treatment; 2D crystal violet absorbance assessed at day 10±2 after treatment; 3D cell count and viability assessed at day 12±2 after treatment.
EEG of therapist and control/sham therapist will be assessed throughout each experiment per protocol.
**DAY TWO:**
- Experiment 3: Dose: 5 minutes; 30 minutes; 15 minutes
- Experiment 4: Dose: 15 minutes; 60 minutes; 30 minutes
Outcomes assessed in both experiments include: pH of the medium and Prestoblue proliferation assay assessed at 0 hour after treatment.
Outcomes assessed in just one experiment: Cell membrane voltage potential dye for the 30-minute dose assessed at 0 hour and 3, 5, and 7 days after treatment.
EEG of therapist and control subject will be assessed throughout each experiment per protocol.
All other procedures remain as outlined on the original protocol.