Evaluation of Nano-Pulse Stimulation (NPS) in Sebaceous Hyperplasia
Phase: NA
Conditions: Skin Lesion, Skin Abnormalities, Sebaceous Hyperplasia
Sponsor: Pulse Biosciences, Inc.

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PROTOCOL 
NANO-PULSE STIMULATION IN SEBACEOUS HYPERPLASIA 
(NP-SH-006) 
MAY 22, 2018 
CONFIDENTIAL 
PMB/5/22/18

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NANO-PULSE STIMULATION IN SEBACEOUS HYPERPLASIA 
(NP-SH-006) 
BACKGROUND 
Sebaceous Hyperplasia (SH) is a common condition that appears as white or lightly 
pigmented bumps or bulges on the skin that occur when hyperactive sebaceous glands 
produce excess oil that pushes up on the skin surface. There are sebaceous glands all 
over the body, so the SH bumps can form almost anywhere, though there are more 
frequently observed and treated when they appear on facial skin. These benign lesions 
SH are more likely to occur in middle-aged and older people, but they can show up at any 
age.  The sebum produced by sebaceous glands is excreted through skin pores and helps 
the skin form a lipid barrier to common environmental stresses. Too much sebum 
production can contribute to several problems, including acne, oily skin, or an oily scalp.
When Sebum can also clog a pore and create and cause selling between the gland and 
the surface of the skin, causing it to swell and form a bump under the skin. Some 
medications and home remedies may reduce the appearance of bumps or reduce sebum 
production, and many patients request cosmetic procedures to remove them.  There is 
currently no permanent cure for sebaceous hyperplasia. 
Several factors increase the likelihood of developing sebaceous hyperplasia. Fair-
skinned people older than 40 tend to develop the condition, especially when their skin 
has frequently been exposed to the sun. Continual sun damage can worsen symptoms 
or cause them to appear earlier. People may be more likely to develop sebaceous 
hyperplasia if they have a family history. People with suppressed immune systems and 
those taking the immunosuppressant medication cyclosporine may have a higher risk of 
developing sebaceous hyperplasia. 
The main symptom of sebaceous hyperplasia is the appearance of small, shiny bumps 
under the skin. A bump can have a slight indentation in the center and a white or yellow 
outer edge. It may be difficult for a non-dermatologist to distinguish the condition from 
acne. However, a whitehead or blackhead will usually have a lifted center, while bumps 
caused by sebaceous hyperplasia are indented. These SH bumps are typically small and 
cause no pain. Many people with oily or combination skin may notice these bumps as 
they age. Bumps may appear on their own or in small clusters. 
A sebaceous hyperplasia lesion is benign, and does not usually require medical 
treatment.  However, many patients choose to have them removed for cosmetic reasons. 
if the bumps are unsightly or embarrassing. Various modalities are available to improve 
the appearance of SH lesion, but a few sessions or applications are often required for full 
removal and prevention of recurrence. Remedies include topical Retinol, facial peels, 
laser therapy, cryotherapy, electrotherapy, photodynamic therapy, excisional surgery, 
antiandrogen medications, and a variety of home/over-the-counter remedies. Pulse 
Biosciences proposes to conduct a clinical evaluation of their Nano-Pulse Stimulation 

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(NPS) device as a modality in the treatment and management of Sebaceous Hyperplasia 
(SH) in patients bothered by their appearance. 
NPS TECHNOLOGY REVIEW 
The NPS medical device is intended to clear the skin of benign, undesired skin lesions 
as an alternative to surgery and other more destructive methods for removing benign 
lesions. The system is designed to deliver a timed series of very low energy, high 
voltage (about the same as the voltage of static electricity) pulses of a time length 
between 100 and 750 nanoseconds (billionths)   of a second). The non-thermal effect on 
tissue takes place in a very shallow depth of skin directly below the sterile treatment 
tip. The device emits significantly less thermal energy than existing laser, electro-
surgery or electro-cautery equipment. 
Extensive in vitro research demonstrates stimulation of a form of delayed programmed 
cell death in a treated area, which is one of the identified mechanisms for observations 
of clinical and histologic changes t o  c e l l s  in multiple studies of in-vivo treated tissue 
in both humans and animals. In pre-clinical studies with over 1000 rats and mice, 
nano- pulsed devices have been demonstrated to reduce or eliminate m a l i g n a n t  
tumors with a clinica lly acceptable margin of safety. A pilot clinical trial of common 
skin cancer lesions was conducted under IRB oversight in 2012 with a similar 
version of the NPS device, with the findings published in Experimental Dermatology. A 
total of ten basal cell carcinoma lesions on the skin of three Subjects were treated 
with a range of settings. Seven of the ten lesions were free of basal cells at the 
conclusion of the study, two partially resolved, and one recurred. No scars were visible 
at the healed sites of any of the successfully treated lesions. 
PRIOR NPS CLINICAL STUDIES  
The NPS device has been successfully deployed in several clinical studies of skin and 
subcutaneous structures.  As of the date of this writing, the following studies have been 
successfully conducted and concluded:  1) A histological study evaluating  use of the NPS 
on healthy abdominal human tissue in a dose-response study; 2) A histological study 
evaluating use of the NPS and comparative devices on healthy abdominal human tissue, 
3) histological study following use of the NPS on healthy facial skin tissue in the pre-
auricular area; 4) A multi-center study in the treatment of Seborrheic Keratosis lesions  
Additional detailed summaries of these studies follows.   
Abdominal Tissue (Pre Abdominoplasty) Studies 
Study Name:  
Nano-Pulse Human Tissue Study 
Study Number: 
NP-ABD-001 
Device Named: 
NPS 
PI Name: 
 
David Kaufman, MD 

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Co PI:  
Michele Martinez, RN 
Study Dates:  
November 2016  
A total of eight Subjects participated in this prospective, single site, non-randomized 
study.  All of the participants were females representing a wide range of Fitzpatrick Skin 
Tone Classifications, and ranging in age from the low 40’s to middle 60’s. 
The study was comprised of two sequential segments. The first segment evaluated the 
serialized use of the test device in demarcated locations on intact abdominal skin 
predetermined for subsequent resection during a standard abdominoplasty scheduled 
at a later date. The primary endpoint of the first segment of the study was to establish 
that the trauma to the tested skin would be minimal over a range of six progressively 
increasing energy levels. The appearance of each intact tested tissue location was 
comparable to that anticipated following any minor surgical excisional procedure. A 
total of 210 NPS applications were tested and analyzed in this study. 
Lidocaine was injected prior to use of the NPS skin exposure.  Subjects reported little or 
no sensation during the treatment session. Subjects further reported little or no discomfort 
after the Lidocaine effect had diminished. There were no adverse events, no reports of 
bleeding, and no scarring noted by the investigator at the end of the 60 day study 
The second segment of the study called for the histological analysis of tissue 
samples prepared from the abdominal section that were resected at the time of the
abdominoplasty. Internal technicians and outside expert dermatopathologists 
evaluated the skin samples using standardized methods of microscopic analysis. This 
second segment did not impact the participating Subject as it occurred after the 
abdominoplasty procedure. Consequently, Pulse Biosciences, Inc. secured IRB 
oversight concentrated on the first segment of the study. It should be noted however that 
the histological analysis to date confirmed that the NPS was successful in achieving its 
intended effect across the range of tested energy settings.  
Abdominal Tissue (Pre-Abdominoplasty) Comparative Studies 
Study Name:  
Nano-Pulse Human Tissue Study 
Study Number: 
NP-ABD-001C 
Device Named: 
NPS 
PI Name: 
 
David Kaufman, MD 
Co PI:  
Michele Martinez, RN 
Study Dates:  
August 2017  
A total of 2 Subjects participated in this prospective, single site, non-randomized study.  
Both of the participants were females representing two different Fitzpatrick Skin Tone 
Classifications, and ranging from the high 20’s to low 40’s in age.   
The primary purpose of the study was to collect tissue samples from healthy human

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resected tissue that had been exposed to various methods of partial tissue ablation prior 
to a planned abdominoplasty resection date. The skin samples were taken from a 
section of abdominal skin that was previously exposed to the effect of the NPS during 
the same time period as two comparator medical devices considered to be the “gold 
standards” for tissue ablation. Histological analysis of the resected abdominoplasty 
tissue was performed on samples taken from sites treated with the 2 “gold standard” 
devices as well as the NPS. 
The study is best described as comprised of two sequential segments. The first 
segment called for the serialized use of the NPS test device and the gold standard 
“comparatives” in specific locations on intact abdominal skin predetermined to be 
resected during a standard abdominoplasty. The impact of the study test device, the 
comparative devices and/or the study process ceased at the time of the 
abdominoplasty. The primary endpoint of the first segment of the study was to 
establish that the apparent trauma to the tested skin was minimal for all devices. 
The appearance of each intact tissue location that was tested with the NPS or the 
two comparative systems was comparable to that anticipated following any minor
surgical excison procedure.  
Lidocaine was injected prior to use of the NPS skin exposure, and one of the 
comparators.  Subjects reported little or no sensation during the multiple treatment 
sessions. Subjects further reported little or no discomfort after the Lidocaine effect had 
diminished in the days following each treatment. There were no adverse events, no 
reports of bleeding, and no scarring in the NPS group noted by the investigator at the 
end of the 60 day study 
The second segment of the study called for the histological analysis of tissue 
samples prepared from the abdominal section that was resected at the time of the
abdominoplasty. Internal technicians and outside expert dermatopathologists 
evaluated the skin samples using standardized methods of microscopic analysis. 
Pre-Auricular Tissue (Pre-Face Lift) Studies 
Study Name:  
Nano-Pulse Pre Auricular Tissue Study 
Study Number: 
NP-PT-003 
Device Named: 
NPS 
PI Name: 
 
James Newman, MD 
Study Dates:  
October 2017  
A total of 3 Subjects participated in this prospective, single site, non-randomized study.  
Two of the participants were females and one was a male representing varying different 
Fitzpatrick Skin Tone Classifications and all were over the age of 50 years of age. 

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The study was compromised of two sequential segments.  The first segment called for 
the serialized use of the test device in several locations on intact facial skin pre-
determined to be resected during a standard face-lift, which is typically a small section in 
front of the ear (the pre-auricular area).  
The primary endpoint of the first segment of the study was to establish that the visually 
apparent trauma to the tested skin would be minimal and the appearance of each intact 
tested tissue location was comparable to the appearance that is anticipated following any 
minor surgical procedure. 
Lidocaine was injected prior to NPS administration to the skin surface.  After the first NPS 
exposure, Subjects returned at varying time intervals prior to their scheduled face lift 
procedure for additional NPS administrations on demarcated areas on each side of their 
face using varying tip sizes and energy settings.  Subjects reported little or no sensation 
during the NPS administrations.  Subjects further reported little or no discomfort after the 
local anesthetic effect had diminished.  There were no adverse events, no reports of 
bleeding and no scarring noted by the Investigator at the end of the study.   
As was noted for the pre-abdominoplasty tissue study the second segment of the pre face 
lift study called for the histological analysis of tissue sample prepared from the tissue 
section that was resected at the time of the face lift.  Internal and outside expert 
histologists were consulted.  See Attachment A for a report authored by Darius R. 
Mehregan, MD, and expert dermatopathologist, based on his independent analysis of the 
biopsy samples from this study of facial skin.   
Following the same model as described above, additional studies were performed 
involving pre-auricular tissue of facial skin in 3 Subjects scheduled for eventual face lift 
procedures.  The sole Investigator for this series (NP-PT-003) was Dr. James Newman, 
MD., Plastic Surgeon, in San Mateo, CA.  As noted above for the pre Abdominoplasty 
study, local anesthetic was administered prior to the NPS skin exposure.  Subjects 
reported little or no sensation during the treatment session. Subjects further reported little 
or no discomfort after the local anesthetic effect had diminished. There were no adverse 
events, no reports of bleeding, and no scarring noted by the PI at the end of the 60 day 
study. 

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Seborrheic Keratosis Study 
Study Name:  
Nano-Pulse Seborrheic Keratosis Study
Study Number: 
NP-SK-002 
Device Name: 
NPS 
PI Names: 
 
James Newman, MD 
George Hruza, MD 
Thomas Rohrer, MD 
Brian Zelickson, MD 
Study Initiation Date: 
May 2017  
The SK Study was a prospective, randomized, open label, multi-site, NSR study 
design where each Subject served as his or her own control.   Principle Investigators 
were identified at each of four participating sites.  A total of 58 Subjects participated.  
Forty eight, 48 (83%) of the participants were females and 10 (17%) were male.  The 
Subjects were primarily Caucasian (95%).  Fifty one, 51 (88%) were classified as Fair or 
Medium using the Fitzpatrick Skin Tone Classifications, and ranged in age from 34 to 74 
years old. 
For each Subject, 4 SK’s were selected that met the study criteria for treatable SK’s.  If 
more than four lesions were present, the Subject stated a preference for which 4 lesions 
would be included in the study.  Photographs of each of the four selected SK’s were taken 
prior to anesthesia or treatment. Local anesthesia w a s  injected at the site of the four 
selected SK’s.  Using a pre-determined randomization method, three of the selected 
SK’s were designated to be treated and one was designated as an untreated control.  The 
test device was deployed to the selected SK's according to a randomized order.   
Subjects returned at prescribed intervals for evaluation of the treated lesion as well as the 
skin around the lesion.  Photographs were taken of each lesion at all intervals. 
Among the 58 Subjects, 58 control lesions were identified and 174 lesions were treated. 
Lidocaine was injected prior to the NPS skin exposure.  Subjects were asked to rate their 
discomfort on a 0-10 point scale where 0=no discomfort.  Subject’s reported little or no 
sensation during the treatment session. Subjects further reported little or no discomfort 
after the local anesthetic effect had diminished.  Ratings were analyzed on a per Subject 
basis and on a per lesion basis.  69% of Subjects rated their discomfort at None, Minimal 
or Mild and when considered on a per NPS administration basis, 68% reported discomfort 
to be None, Minimal or Mild.   
Each lesion area was routinely examined at each study visit for the presence of bleeding, 
scabbing, crusting, erythema, swelling, exudate, indurations, etc. Evidence of blood 

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tinged oozing was noted at the pre-treatment site of some of the injected Lidocaine 
anesthesia infiltrations. No frank bleeding was noted at any time.  
 Minor scabbing, crusting and flaking were noted in the early post treatment visits but 
lessened over time in the natural course of skin healing.  There were no adverse events 
and no complications.   
At the final 106 day post treatment visit, using a standard scale, the investigators rated 
the clearance level of each lesions and the appearance of the residual skin.  The Subjects 
rated their satisfaction with the appearance of each lesion.  Investigators were asked to 
observe and rate the lesion clearance using the following scale: 0 = Clear; 1= Mostly 
Clear; 2= Partially Clear and 3= Not Clear.  Each treated lesion was rated at each 1-
month post; 60 days post; 90 days post and 106 days post treatment. The treated lesion 
was considered improved when at least partial clearance was observed. Combining all 
three clearance levels, 87% of the lesions had improved at 1 month and 93% showed 
improvement at 106 days post treatment.  
Investigators used a comparable scale to evaluate and report the appearance of the skin 
within the margins of the NPS location and adjacent to each lesion location. The scale 
provided included: 0 = Clear; 1 = Hyper-pigmentation; 2 = Hypo-pigmentation or 3 = Other 
as rating options.  32% of the surrounding skin sites were recorded as Clear.   
Some Hyper-pigmentation was noted for 60% of the treated lesions surrounding skin. 
Hypo-pigmentation was seen in 2% and 6% of the ratings were recorded as Other. 
Subjects were asked to rate their level of satisfaction with the outcome for each of their 
treated lesion.  The rating scale included: Satisfied; Mostly Satisfied, Partially Satisfied, 
Dissatisfied and Highly Dissatisfied.  Subjects reported being at least partially Satisfied 
for 93%, (n=161) of the treated lesions. 
An Independent Photographic Review was conducted following the close of the study. 
Each reviewer rated photographs of 196 lesion sets taken at the pre-treatment and 106 
day visits respectively. This review was conducted in accordance with established 
standards for validity and reliability.  The rating scale of the independent review was the 
same one utilized by the Principle Investigators who participated in the study.  Prior to its 
use in the Independent Photographic Review, the Rating Scale was validated. Inter-rater 
Reliability was established at 82% and Intra-rater Reliability was established at 83%.  
Success Criteria for purposes of the Independent Photographic Review was set as an 
improvement of at least 2 grades on a 4 point rating scale.  71% of NPSs met the success 
criteria.   
PRIOR CLINICAL STUDIES CONCLUSIONS 
A combined total of 642 NPS applications have been delivered to 71 adult Subjects. 
Anatomic locations included abdomens, backs, arms, legs and faces.  Discomfort was 
managed with localized Lidocaine.  For 58 of the Subjects a favorable therapeutic 
outcome was established in a significant majority.  Side effects consisted of relatively 

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minor reactions consistent with routine wound healing.  No complications or adverse 
events were reported. 
PROPOSED SEBACEOUS HYPERPLASIA STUDY 
Study Design 
Sebaceous Hyperplasia (SH) is a common condition that appears as white or lightly 
pigmented bumps or bulges on the skin that occur when hyperactive sebaceous glands 
produce excess oil (sebum) that pushes up on the skin surface. These benign SH lesions 
are more likely to occur in middle-aged and older people, but they can show up at any 
age. There is currently no permanent cure for sebaceous hyperplasia. 
A sebaceous hyperplasia lesion is benign and painless, and does not usually require 
medical treatment.  However, many patients choose to have them removed by physicians 
for cosmetic reasons, especially if the bumps are viewed as unsightly or embarrassing. . 
Various modalities are available to improve the appearance of SH lesions, but a few 
sessions or multiple applications are often required for full removal and prevention of rapid 
recurrence. Remedies include topical Retinol, facial peels, laser therapy, cryotherapy, 
electrotherapy, photodynamic therapy, excisional surgery, antiandrogen medications, 
and a variety of home/over-the-counter remedies.  
The proposed SH Study using NPS treatment of individual lesions is a prospective, open 
label, multi-center non-significant risk study design where each Subject serves as his or 
her own control.  A Principle Investigator (PI) will be identified at up to 4 participating study 
centers.  A total of up to 75 Subjects will be consented and enrolled with an anticipated 
enrollment expected to be uniformly distributed across sites. The PI will consider the study 
inclusion and exclusion criteria and invite appropriate patients to participate in the study.   
The intended treatment device is described as the Nano-Pulse Stimulation (NPS) device 
and has been used in a similar manner in prior studies of skin lesion clearance. For the 
proposed SH lesion study, the skin contacting device component referred to as the “tip” 
will be generally be 1.5mm x1.5mm or 2.5 mm x 2.5mm in size with energy settings in the 
mid-range of previously tested energy levels. Local anesthetic in the treatment area will 
be used as needed to manage potential treatment discomfort.
Each Subject may present with a minimum of 2 and up to 4 designated SH lesions, with 
one lesion designated as an untreated control by a randomized method.  All treated 
lesions will receive at least one NPS treatment.    At 30 days post-treatment, if any treated 
lesion that is rated by the investigator as Not Clear or Partially Clear on a study 
standardized scale will be eligible for a second treatment to enhance lesion clearing. If a 
second treatment is indicated for some or all of the lesions the second treatment will occur 
at 30 days after the initial treatment. Total study time for these Subjects with any lesion 
requiring two treatments will be 90 days.   Subjects with all treated lesions requiring only 
one treatment will complete the study in 60 days.   

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No significant risk is posed by either the use of the NPS or the study process. While safety 
can never be taken lightly, this study is not meant to measure monitor or analyze any 
significant disease or disorder for which medical treatment is mandatory.     
See additional information regarding Non-Significant Risk statement below. 
Non-Significant Risk Statement
The proposed study fits the criteria for a non-significant designation for the 
following reasons: 
• 
Subjects will be recruited to participate in the study on a voluntary basis.  
Potential participants will be recruited directly from the Investigator’s 
population of patients who are candidates for removal of their SH lesions. 
• 
Participation in the study takes no more than approximately 1 hour for 
each of the maximum of 5 study visits.
• 
Localized anesthesia will be used to control discomfort at the time of any 
NPS device applications. 
• 
After the NPS device is deployed and the anesthesia is dissipated, there
may be mild, localized discomfort at the site of the treatment. A small scab 
or crust may develop. This occurrence is transient and should resolve 
without intervention. 
• 
Prior studies support the proposal that the NPS use represents minimal risk.  
• 
There is no loss of privacy as no study records will be viewed or retrieved
by anyone other than the study team members. 
• 
Protected information will not be captured. 
• 
Subject identity is protected by a coding system to de-identify them. 
Study Objectives 
 Per SH Lesion:  NPS treated lesions will be rated via a previously used scale for 
similar NPS studies. At the Final Study Visit, the PI will rate each lesion in 
comparison to an image of that same, pre-treated lesion.  At least 50% of the NPS 
treated lesions will be rated as “CLEAR” or “MOSTLY CLEAR”.  
 Per Subject:  At the Final Study Visit at least 50% of the Subjects will present with 
50% of their lesions rated as “CLEAR” or “MOSTLY CLEAR”.  
 No AE’s or complications will be reported specific to the NPS device or the study 
process. 
Success/Failure Criteria 
Study outcome measures will be applied at every study visit and success/failure criteria 
will be applied at 60 days after the first NPS treatment for lesions treated one time or 90 
days after the first NPS treatment for those lesions treated twice. Success will be 
determined as described in the preceding paragraph regarding Study Objectives.  In 
patients with both single treatment lesions and multiple treatment lesions, all treated 
lesions will be evaluated and images captured at all visits, regardless of number of 
treatments

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Investigator/Site Selection 
Investigators/Site selection and qualification will be initiated upon IRB approval of the 
protocol and consent provided in this submission.  Several of the potential PIs have 
participated in prior studies using the NPS device.   
Subject Selection 
Up to 75 Subjects with 2-5 qualifying SH lesions who are interested in removal of those 
SH lesions and meet the full study criteria will be consented to participate in a study to 
evaluate the outcome using the NPS device.  The specific study Inclusion and Exclusion 
criteria are outlined in the following paragraphs.   
Inclusion Criteria:   (Must be a “YES” Response)
• Willing to sign the Informed Study Consent 
• English speaking/reading and/or translation is available for all visits 
• Males or females 
• At least 18 years of age but less than 71 years of age 
• Presents with at least 2 and up to 5 clinically visible SH lesions 
• Understands that 1 lesion will remain untreated to act as a reference 
• Lesions must measure no greater than 2.5 x 2.5 at the outside margin 
• Wishes to have at least 1 and perhaps 2 NPS treatments to each study lesion  
• Selection of the non-treated reference lesion will be randomly identfied 
• Willing to return to the PI’s office for 4 or 5 total  study visits at specified 
intervals over 60 or 90 days 
• Agrees to photographic or other image capture methods of both the treated and 
untreated lesions.
• Agrees to avoid any other treatment to the NPS treated SH lesions and to the 
untreated SH lesion until the end of the NPS study
• Understands that their personal identity will be masked and all availability of 
images will be restricted to the study team.  
• Has no evidence of active infection in the designated tissue prior to treatment 
and reports no infection within 90 days 
• Is not allergic to Lidocaine or Lidocaine-like products 
• Investigator determines that the patient is in good general health and is unlikely 
to be subject to risks from the treatment 
Exclusion Criteria: (Must be “NO” Responses) 
• Presence of Implantable electronic devices that cannot be removed. e.g., 
pacemaker or automatic defibrillator 
• Taking medications prescribed for cardiac arrhythmia at any time within 6 
months prior to exposure to the NPS device   
• SH lesions are located within the eye orbit or on the nose  

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• Active infection or history of infection in designated test area within 90 days  
prior to study initiation  
• Use of oral steroid and/or retinoid use within the last 12 months  
• Prior physician ordered treatment to the identified SH lesions targeted for the 
study which occurred within 6 months prior to study start 
• Is known to be immune-compromised and/or received  immunosuppressant 
therapy within 6 months prior to study start 
• Taking blood thinning medications 
• Has Insulin dependent diabetes 
• Is known to be pregnant or lactating female 
STUDY PROCESS 
The following is an example of the Subject related activities to be completed 
for each Subject in the study. The exact order may differ depending on 
Subject/Pl schedules and preferences. 
Activities prior to or on same day as study enrollment and occur on same day 
as treatment visit: 
 
Evaluation for Inclusion/Exclusion criteria 
 
Sign the consent form prior to any study activities 
 
Receive a copy of the signed consent form 
Activities on 1st Study Visit Day-NPS Treatment Day 
• 
Up to 5 SH lesions that meet the study criteria are selected for inclusion in 
the study.  Specifically…. 
• 
if 2 SH lesions are qualified, 1 will be treated and 1 will be reference  
• 
If 3 SH lesions are qualified, 2 will be treated and 1 will be reference 
• 
if 4 SH lesions are qualified, 3 will be treated and 1 will be reference 
• 
if 5 SH lesions are qualified, 4 will be treated and 1 will be reference 
• 
Using a randomization method the selected SH lesions will be designated 
to be treated and one will be designated as an untreated control  
• 
Images of each of the selected lesions will be taken prior to anesthesia or 
treatment 
• 
Local anesthesia will b e  applied to all study  selected SH lesions   
• 
The lesions to be entered into the study will be numerically labeled. Those 
lesions to be treated vs the reference lesion (non-treated) will be randomly 
selected.  
• 
A light bandage and any PI recommended dressing will be applied
• 
Subject will be discharged 

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Activities on 2nd Study Visit Day – 5 Days post NPS treatment 
• Images of each of the treated lesions and one non-treated lesion will be 
taken  
• Each lesion will be clinically assessed and the appearance will be rated 
according to specified criteria.  
• Any adverse events will be identified and documented. 
Activities on 3rd Study Visit Day – 30 Days post NPS treatment 
• Images of each of the treated lesions and one non-treated lesion will be 
taken  
• Each lesion will be clinically assessed and the appearance will be rated 
according to specified criteria  
• If any lesions are rated other than Clear or Mostly Clear, a 2nd NPS treatment 
may be delivered to that previously treated lesion 
• Any adverse events will be identified and documented 
Activities on 4th Study Visit Day – 60 Days post NPS treatment #1 
• Images of each of the treated lesions and one non-treated lesion will be 
taken 
• This is Final Study visit for any patient for whom all  lesions were treated 
one time  
• Each lesion will be clinically assessed and the appearance will be rated 
according to specified criteria  
• Any adverse events will be identified and documented 
Activities on 5th Study Visit Day – 90 Days post NPS treatment #1  
• Images of each of the treated SH lesions and one non-treated SH lesion will 
be taken  
• This is Final Study visit for any treated lesion  
• Each lesion will be clinically assessed and the appearance will be rated 
according to specified criteria  
• Any adverse events will be identified and documented 

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SH Study Follow-up Visit Schedule 
Study Visit 
Interval 
#1 
NPS TX*
#2 
5 days post 
NPS TX
#3 
30 days post 
NPS TX
#4 
60 days post 
NPS TX #1
#5 
90 days post 
NPS TX
Study Visit 
Description 
TX  #1 
Wound 
Check 
Wound 
Check 
TX  #2  
(if applicable) 
Wound 
Check  
Final visit for 
all lesions 
treated only 
one time  
Wound 
Check  
Final visit for 
all lesions 
treated more 
than one 
time
Visit Range 
(# of Days 
Post First  
NPS TX) 
4-8 days 
28-35 days 
57-67 days 
87-97 days 
*TX = Treatment 
SUBJECT COMPENSATION
Each Subject will be compensated for any inconvenience that study participation 
may represent. Specifically, each Subject will make 4-5 total visits to the PI’s office 
(approximately 4-5 hrs. total time).  In return for this time commitment, the Subject will 
receive $200 per treatment visit, $100 per check-up visit and $200 for the Final Visit. An 
additional $400 bonus will be paid for completing all of the scheduled visits. The total 
amount of compensation will be up to $1000 to those Subjects who receive one NPS 
treatment and $1200 to those Subjects who receive more than one NPS treatment.  
STUDY BENEFITS
A volunteer for the proposed study is someone who has elected removal of SH lesions.  
Such scars are typically removed via surgical excision, intra-scar steroidal injections or 
systemic steroids. Participation in the study offers another technique to accomplish the 
same thing as the typical tools.  Experience with the NPS to date offers some confidence 
that the scars may be lessened although no assurances will be offered. The information 
learned may contribute to the ultimate use of a more safe and effective device 
available to treat benign and non-benign lesions in future patients. 
STUDY/NPS DEVICE RISKS
The Investigators for the study are experienced physicians. Use of the NPS poses 
very minimal risk, including scabbing, minor skin pigment changes or scarring and minor 
discomfort which can be further minimized by covering the spot with a small bandage. 

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Typical wound complications such as infection, bleeding and discomfort which requires 
prescription medication for relief are not anticipated. If they should occur they would 
be considered an unanticipated adverse event and managed as such.  If the Subject 
has any questions or concerns about their medical condition or if an unforeseen event 
should occur the Subject will be advised to contact the PIs or their designate.  
SITE TRAINING PLANS
Training to be conducted can be divided into two pertinent categories. First is training 
on use of the NPS from a technical and clinical perspective. The PIs and their 
designated assistants will be instructed in aspects of set up, application, management 
and maintenance of the NPS. Those persons conducting the training will be Pulse 
Bioscience technical experts in this area of expertise. A select group of these experts 
will be present during the testing sessions to provide support as needed. Training will 
include a dry run. The second category is training in the clinical study requirements 
and processes. The study team members at the Study Site will be trained via the 
pertinent documents and files as well as the planned logistics to perform the study
according to the protocol and applicable IRS, GCP and Pulse requirements. 
INSTITUTIONAL REVIEW BOARD OVERSIGHT
The following Institutional Review Board will be asked to review and approve the
proposed study: 
Biomedical Research Institute of America 
P.O. Box 600870 
San Diego, CA 92160
T - 619 282 9997 
F - 619 282 9998 
If the Subject has any questions or concerns about the study process, they will be 
advised to contact Biomed at 619-282-9997 
STUDY MANAGEMENT AND MONITORING PLAN
Pulse Biosciences, and/or their designated representatives are solely responsible for 
the proposed study.  Pulse will take the necessary steps to assure that the study is 
conducted in accordance with all regulating authorities as well as applicable Standard 
Operating Procedures.  The Study Monitor will oversee the conduct of the study on a 
continual basis. Minimal Monitoring visits will be conducted at the following intervals: 
Site Qualification Visit, Study Initiation Visit, Interim Study Visit and Study Close-Out Visit. 

16 
Attachment A  
A Summary Histology Report of the Nano-Pulse Preauricular Facial Skin Study 
(NP-PT-003)
 in a Pre-Excised Human Facial Tissue Model 
Darius R. Mehregan, MD 
Microscopic samples of preauricular facial tissue from three Subjects treated with a novel 
non-thermal energy device that delivers short low energy, high voltage pulses to a 
controlled tissue volume have been reviewed in a detailed histologic analysis.  The 
treatment intervals of the targeted normal preauricular facial tissue ranged from 15 to 60 
days prior to a previously scheduled facelift procedure in which the treated tissue was 
demarcated two months prior to excision.  Six treatment energy levels are reviewed per 
patient and are evaluated at one of the three timepoints prior to their surgery: 60 days, 30 
says and 15days. Various tissue staining techniques intended to identify specific cellular 
changes and tissue morphology were utilized in the histologic analysis to characterize the 
tissue responses and the subsequent recovery processes. The findings for each of the 
energy levels tested were compared to normal control punch biopsies in the same patient. 
Epidermal Changes:  Review of the changes of the entire thickness of the 
epidermal/dermal layer and subcutaneous fat from day 15 through day 60 was performed.  
The primary changes due to the non-thermal energy exposure were observed in the 
treated skin within the epidermal layer of skin.   
 In many patients, hair follicles and eccrine glands within the dermal layer of the skin were 
also visible for histologic review.  In specimens in which hair follicles and eccrine ducts 
were visible, there was no apparent effect to the adnexal structures in any of the samples 
observed.  This is consistent with previous histologic analysis of tissue exposed to this 
technology, which showed partial healing seen at 5 days and complete recovery by 15 
days.  Eccrine ducts often showed focal squamous metaplasia, a sign of re-
epithelialization.  The epidermal layer had returned to normal in all cases with a normal 
thickness and preservation of rete ridge pattern similar to controls.  
The samples at the day 15 timepoints showed full re-epithelialization of the epidermis. 
The rete ridge patterns which often show flattening with chronic photoaging, also showed 
patterns similar to control specimens. Evidence of epidermal inflammation such as 
spongiosis or exocytosis of lymphocytes was not observed.  
The samples at the day 30 timepoints showed similar changes to the 15-day samples. 
The samples at the 60 days timepoint showed the epidermis, hair follicles, and eccrine 
glands had all completely returned to normal.   

17 
The epidermal thickness and rete ridge pattern was similar to control specimens. 
Distribution of eccrine glands and ducts as well as hair follicles were comparable to control 
samples.  PT-002 showed evidence of follicular epithelial spongiosis and peri-follicular 
inflammation at day 15 which resolved by day 30. 
Alterations of dermal collagen: Alterations in the dermal collagen observed were 
minimal, with no evidence of thermal injury.  In some tissue samples exposed to treatment 
levels TL25.008, TL25.010 and TL25.012 there was a mild effect on the elastin fibers, as 
there was evidence of a loss of elastic fibers in the papillary dermis.  In one of these 
patients (PT-003) there is also some flattening of the dermo-epidermal junction which 
improved by day 60. 
Elastic tissue staining: Elastic tissue remained intact in the vast majority of patients.  In 
tissue samples exposed to treatment levels TL25.008, TL25.010 and TL25.011 there was 
occasional slight decrease in elastic fibers. Minimal effects were noted on the dermis. 
This predicts a very low risk of scarring. The risk is greatest in treatment level TL25 011 
and 012 (Evident in PT-003 30 days). 
Melanocytic density:  The number of melanocytes was observed using a MITF 
immunostain. The number of melanocytes is counted in 3 different areas of 1mm and 
averaged. Staining for melanocytes shows the number of melanocytes return to normal 
density, comparable to the control specimens for most energy levels tested. The two 
highest energy levels show a slightly lower count of melanocytes compared to controls, 
which may be indicative of a slower recovery of melanocytes. The rapid return of the 
melanocytic density to levels comparable to control should be consistent with a relative 
normalization of skin pigmentation over time.  
Inflammation: Overall, the degree of dermal inflammation was minimal compared to 
other thermal or physical methods of intentionally damaging surface epidermal tissue.  
There is a small amount of inflammation seen at day 30 for some tissue exposed to 
treatment levels TL25.008, TL25.010 and TL25.012, however the amount of inflammation 
appears to be sparse, primarily perivascular and focally perifollicular.  The samples 
examined showed no evidence of fibrin deposition in the blood vessels to suggest 
vascular injury. 
Presence of Macrophages: Macrophages containing melanin were sparse but present 
in the papillary dermis. These were present at all treatment levels but are few in numbers. 
Fat: There was no effect on the subcutaneous fat at any of the timepoints evaluated. 

18 
Summary:  The novel method of using low energy, high voltage nano-pulse stimulation 
on facial skin and subcutaneous tissue at nine different energy settings was observed to 
lead to a predictable recovery of the epidermal layer of facial skin at all energy levels at 
all timepoints observed.  The lack of observed effect on dermal collagen suggest that the 
effect is non-thermal, with a relatively low level of inflammation.    This lack of inflammatory 
effect is consistent with preservation of fibroblasts, elastic tissue, and melanocyte 
recovery.  The transient effect on deeper cellular structures in the dermis suggest an 
affinity of Nano-pulse energy for highly cellular tissue, and a sparing effect on the less 
cellular connective tissue of the collagen layer. 
Darius R. Mehregan, MD 
Dermatologist and Dermatopathologist 
Dearborn, Michigan