Transcranial Near-Infrared Light in Healthy Subjects: a Cerebral Blood Flow Study With Diffuse Correlation Spectroscopy
Phase: NA
Conditions: Cognitive Change
Sponsor: Massachusetts General Hospital

 
 
 
Version: 8/15/2017 
 
 
 
Page 1 of 39                                                    C O N F I D E N T I A L                                                
 
Confidential Information 
The information contained in this protocol is confidential and is intended for the use of 
clinical Investigators.  It should not be copied by or distributed to persons not involved 
in the clinical investigation of the Litecure laser device , unless such persons are 
bound by a non-disclosure agreement with Litecure, Inc. 
 
 
 
 
 Transcranial Near-Infrared Light in Healthy Subjects: a 
Cerebral Blood Flow Study with Diffuse Correlation 
Spectroscopy (NIR-flow). 
 
 
Study Sponsor 
Paolo Cassano, MD PhD 
Principal Investigator 
Massachusetts General Hospital 
1 Bowdoin Sq. 6th Floor 
Boston, MA 02114 
 
 
 
 
 
 

 
  
 
                          Page 2 of 39                                                    C O N F I D E N T I A L                                           
.    
 
Table of Contents 
1 STUDY SUMMARY ............................................................................................................................... 4 
2 LIST OF ABBREVIATIONS ................................................................................................................. 6 
3 INTRODUCTION ................................................................................................................................... 7 
3.1 
SUMMARY OF PRIOR CLINICAL EXPERIENCE ........................................................... 7 
3.2 
OVERVIEW OF LITECURE  SYSTEM…………………………………………...…..8 
3.3 
OVERVIEW OF THE DCS DEVICE.......…………………………………………...….9 
3.4 
Overview of the EEG Device………………………………………………….....10 
3.5       Overview of Rationale for DCS, EEG and Cognitive Task....................................11 
4 INVESTIGATIONAL PLAN ............................................................................................................... 12 
4.1 
RATIONALE ............................................................................................................ 12 
4.2 
STUDY DESIGN ...................................................................................................... 12 
4.3 
STUDY POPULATION .............................................................................................. 13 
4.3.1 Population Description ........................................................................................................ 13 
4.3.2 Use in Female Subjects of Childbearing Potential .............................................................. 14 
4.3.3 Criteria for Enrollment ........................................................................................................ 14 
4.4 
SUBJECT NUMBERING ............................................................................................ 14 
4.5 
SAMPLE SIZE .......................................................................................................... 14 
4.6 
SUBJECT CONFIDENTIALITY ................................................................................... 14 
4.7 
STUDY VISITS ........................................................................................................ 14 
4.8 
DETAILED STUDY VISITS ....................................................................................... 15 
4.8.1 Screening Visit (Week 1) ...................................................................................................... 15 
4.8.2 TLT-DCS Visits – Visits 2-4(Weeks 2-4) .............................................................................. 16 
4.8.3 Follow-Up Visits – Visit 5  (Week 5)…………...................................................................... 17  
4.8.4 Termination .......................................................................................................................... 18 
4.9 
SCALES & FORMS .................................................................................................. 18 
4.10 
COGNITIVE TESTS ................................................................................................... 19 
4.11      EEG RECORDING…………………………………………………………………20 
4.12 
DATA MANAGEMENT .............................................................................................. 21 
4.13 
TRAINING ............................................................................................................... 21 
4.14 
BLINDING OF THE STUDY ........................................................................................ 21 
4.15 
COMPENSATION ..................................................................................................... 22 
4.16 
ACCOUNTABILITY/INVESTIGATIONAL PRODUCT CONTROL ..................................... 22 
4.17 
CLINICAL ADVERSE EVENTS ................................................................................... 22 
4.17.1 Overview and Definitions .................................................................................................... 22 
4.17.2 Safety Monitoring ................................................................................................................ 23 
 
4.17.3 Reporting Procedures for All Adverse Event. ......................................................................24 
 
4.17.4 Serious Adverse Events.........................................................................................................26 
 
4.17.5 Deaths...................................................................................................................................26 
 
4.17.6 Withdrawals for Adverse Events...........................................................................................26 
4.18     MEASURES TO ASSURE SUBJECT'S SAFETY............................................................26 
4.19 
SUBJECT'S DISPOSITION CRITERIA...........................................................................27 
 
4.19.1 WITHDRAWAL FROM STUDY....................................................................................................27 
 
4.19.2 LOST TO FOLLOW-UP..............................................................................................................28 
4.20 
RISK AND BENEFITS OVERVIEW..............................................................................28 
 
4.20.1 POTENTIAL ADVERSE EVENTS.................................................................................................28 
 
4.20.2 BENEFITS...............................................................................................................................29 

 
  
 
                          Page 3 of 39                                                    C O N F I D E N T I A L                                           
.    
 
 
5 DATA ANALYSIS METHODS ........................................................................................................... 30 
5.1 
GENERAL CONSIDERATIONS .................................................................................. 30 
5.2 
SPECIFIC PRIMARY AIM ......................................................................................... 30 
5.3 
SAMPLE SIZE .......................................................................................................... 30 
5.4 
SUBJECT DEMOGRAPHIC AND BASELINE CHARACTERISTICS .................................. 30 
5.5 
EFFICACY ANALYSES FOR PRIMARY AIMS ............................................................. 30 
5.6 
SAFETY ANALYSES ................................................................................................ 30 
5.7 
ADDITIONAL SAFETY ANALYSES ........................................................................... 31 
6 ETHICAL REVIEW & REGULATORY CONSIDERATIONS ...................................................... 32 
6.1 
ETHICAL REVIEW ................................................................................................... 32 
6.2 
REGULATORY CONSIDERATIONS ............................................................................ 32 
6.3 
MONITORING PROCEDURES .................................................................................... 32 
6.3.1 Training of Study Site Personnel ......................................................................................... 33 
6.4 
INFORMED CONSENT .............................................................................................. 33 
6.5 
PROTOCOL ADHERENCE ......................................................................................... 33 
6.6 
DATA COLLECTION ................................................................................................ 33 
6.7 
RECORD RETENTION .............................................................................................. 34 
6.8 
CLINICALTRIALS.GOV ............................................................................................ 34 
7 REFERENCES ................................................................................................................................... 35 
8 LIST OF PROTOCOL ATTACHMENTS.......................................................................................... 38 
8.1 
PROTOCOL ATTACHMENT 1 – TIME TABLE OF STUDY PROCEDURES...................... 38 
8.1 
PROTOCOL ATTACHMENT 2 – SCHEMA ............................................................... 39 
 
 

 
  
 
                          Page 4 of 39                                                    C O N F I D E N T I A L                                           
.    
 
Transcranial Near-Infrared Light in Healthy Subjects: a 
Cerebral Blood Flow Study with Diffuse Correlation 
Spectroscopy (NIR-flow). 
 
 
1. Study Summary 
Protocol #: 
 
 
 
Study Purpose: The purpose of this study is: 
 To assess the change in cerebral blood flow induced by the 
Transcranial Continuous and Pulse Near-Infrared Light 
Therapy in healthy subjects 
 To correlate with cognitive performance the change in cerebral 
blood flow induced by the Transcranial Continuous and Pulse 
Near-Infrared Light Therapy in healthy subjects  
 To correlate with skin pigmentation the change in cerebral 
blood flow induced by the Transcranial Continuous and Pulse 
Near-Infrared Light Therapy in healthy subjects 
 To correlate with brain electrical activity the change in 
cerebral blood flow induced by the Transcranial Continuous 
and Pulse Near-Infrared Light Therapy in healthy subjects 
 To assess the safety and tolerability of the Transcranial 
Continuous and Pulse Near-Infrared Light Therapy in healthy 
subjects 
 
 
 
Trial Design: 
In essence, this study consists in three sequential sessions of Diffuse 
Correlation Spectroscopy (DCS), each one week apart, to test three 
different modes of intervention with Transcranial Light Therapy 
(TLT). Healthy subjects will receive first a session with continuous 
light (c-TLT), then a session with sham (s-TLT) and one session with 
pulse light (p-TLT). Before and after each session, subjects will 
undergo DCS, which will allow measuring the impact of the 
intervention on cerebral blood flow. We expect that out of 30 subjects 
enrolled 10 subjects will be eligible to complete the study. The study 
will be conducted in single blind.  
 
 
Study 
Population: 
Subjects eligible for study participation will be healthy by Structured 
Clinical Interview for Diagnostic Statistical Manual-IV (SCID) 
criteria.  Subjects will be between 18-70 years of age (have not had 
71st birthday) on the date of screening. Subjects who will be permitted 
into the study include those who meet all the inclusion criteria and 
have none of the exclusion criteria.   

 
  
 
                          Page 5 of 39                                                    C O N F I D E N T I A L                                           
.    
 
 
 
Duration of 
Participation: 
Subjects will be followed in the study for 5 weeks. 
 
Endpoints: 
 
Engagement of cerebral blood flow – The effect of TLT on 
cognition will be assessed before and after each session (c-TLT, s-
TLT and p-TLT) by measuring the Diffuse Correlation Spectroscopy 
(DCS) signal.   
Engagement of cerebral electrical activity – The effect of TLT on 
brain electrical waves will be assessed before and after each session 
(c-TLT, 
s-TLT 
and 
p-TLT) 
by 
measuring 
the 
Electro-
Encephalographic (EEG) signal.   
Engagement of cognitive circuitry – The effect of TLT on working 
memory will be assessed immediately after each session by n-back 
task.   
Safety - The safety endpoints will evaluate any reported adverse 
events within one week from each sessions of TLT.     
 
 
 
Entry Criteria: Inclusion Criteria 
 
1. Subject’s age at screening will be between 18 and 70 years old 
(inclusive). 
2. Women of child-bearing potential must use a double-barrier 
method for birth control (e.g. condoms plus spermicide) if 
sexually active.   
3. Subject Informed Consent obtained in writing in compliance 
with local regulations prior to enrollment into this study.  
4. The subject is willing to participate in this study for at least 5 
weeks.  
 
Exclusion Criteria:  
 
1. The subject is pregnant or lactating.  
2. The subject is on any psychotropic medication. 
3. Psychotherapy is exclusionary unless the subject has had at least 
8 weeks of treatment prior to the screening visit. 
4. Any current psychiatric disorder (per SCID assessment) 
5. Substance or alcohol dependence or abuse in the past 6 months. 
6. History of a psychotic disorder or psychotic episode (current 
psychotic episode per SCID assessment). 
7. Bipolar affective disorder (per SCID assessment). 
8. Unstable medical or neurological illness, defined as any illness 
which is not well-controlled with standard-of-care medications 
(e.g., insulin for diabetes mellitus, HCTZ for hypertension). 
9. Suicidal or homicidal ideation as determined by SCID screening. 

 
  
 
                          Page 6 of 39                                                    C O N F I D E N T I A L                                           
.    
 
10. The subject has a significant skin condition (i.e., hemangioma, 
scleroderma, psoriasis, rash, open wound or tattoo) on the 
subject’s scalp that is found to be in proximity to any of the 
procedure sites.  
11. The subject has an implant of any kind in the head (e.g. stent, 
clipped aneurysm, embolised Arteriovenous Malformation 
(AVM), implantable shunt – Hakim valve). 
12. Any use of light-activated drugs (photodynamic therapy) within 
14 days prior to study enrollment (in US:  Visudine (verteporfin) 
– for age related macular degeneration; Aminolevulinic Acid- 
for actinic keratoses; Photofrin (porfimer sodium) – for 
esophageal cancer, non-small cell lung cancer; Levulan 
Kerastick (aminolevulinic acid HCl) – for actinic keratosis; 5-
aminolevulinic acid (ALA)- for non-melanoma skin cancer) 
13. Recent history of stroke (90 days). 
14. Personality traits that rend the subject unsuitable for the study, 
based on the investigators’ clinical judgment.  
 
 
2. List of Abbreviations 
ALA = 5-aminolevulinic acid 
ASQ = Anxiety Symptoms Questionnaire 
ATP = adenosine triphosphate 
AVM = Arteriovenous Malformation 
CADSS=  Clinician Administered Dissociative States Scale 
C-SSRS = Columbia-Suicide Severity Rating Scale 
CFR = Code of Federal Regulations 
CHRT = Concise Health Risk Tracking 
CATSD = Center for Anxiety and Traumatic Stress Disorders 
EEG = electroencephalogram 
FDA = Food and Drug Administration 
LED = Light Emitting Diode 
HAM-D-17 = Hamilton Depression rating scale 
MOCA  Montreal Cognitive Assessment 
MDD = Major Depressive Disorder 
MGH = Massachusetts General Hospital 
NEURO-QOL = Quality of Life in Neurological Disorders  
TSRQ = TLT Self-Report Questionnaire 
OSHA = Occupational Safety and Health Administration 
PANAS = Positive and Negative Symptoms Questionnaire 
PBQ = Perceptions of Blinding Questionnaire  
PI = Principal Investigator at the site 
SAE = Serious Adverse Event 
SAFTEE-SI = Systemic Assessment for Treatment Emergent Events 
SCID = Structured Clinical Interview for Diagnostic Statistical Manual-IV 

 
  
 
                          Page 7 of 39                                                    C O N F I D E N T I A L                                           
.    
 
SDQ = Symptoms of Depression Questionnaire 
SFI = Sexual Functioning Inventory 
TLT = Transcranial Light Therapy 
 
 
 

 
  
 
                          Page 8 of 39                                                    C O N F I D E N T I A L                                           
.    
 
3. Introduction   
Transcranial Light Therapy (TLT) with near-infrared light has recently emerged as a 
potential new antidepressant treatment in both animal models 1 and human studies 2–4. TLT 
consists of delivering invisible near-infrared radiation (NIR), or red light  to the scalp of the 
subject, which penetrates the skull and modulates function of the adjacent cortical areas of 
the brain. Red and/or near-infrared light appears to increase cellular brain metabolism and 
neuroplasticity, and to modulate endogenous opioids, while decreasing inflammation and 
oxidative stress 5–7. TLT has been shown to penetrate deeply into the cerebral cortex 8–10, to 
modulate cortical excitability 11 and improve cerebral perfusion 10 and oxygenation 12. 
Studies have suggested that it can significantly improve cognition in healthy subjects 13–15, 
and in subjects with traumatic brain injury 16–18. The safety of TLT has been studied in a 
sample of  1,410 acute stroke subjects, with no significant differences in rates of adverse 
events between TLT and sham exposure 19–21.  Converging evidence from uncontrolled 
studies suggests both an antidepressant and anxiolytic effect of TLT in subjects suffering 
from Major Depressive Disorder (MDD) 2–4. It is critically important to understand the 
biological targets of TLT and we hypothesize that the improvement in cerebral brain flow 
is one of the underlying mechanism for the boost in bioenergetic metabolism leading to 
therapeutic effects, such as enhancement of cognitive functioning.   
3.1 Summary of Prior Clinical Experience 
 
We conducted two studies on the safety and efficacy of TLT delivered to the forehead 
(prefrontal cortex) twice a week in subjects with MDD.  
 
ELATED-1 was  a proof of concept study of 3 weeks of TLT (Neurothera - class IV 
laser, wavelength 808 nm; irradiance 700 mW/cm2; fluence of 84 J/cm2; total energy 
2.4 kJ per session for 6 sessions) (6). All subjects (n=4) were treated with TLT. 
Baseline mean Hamilton Depression rating scale (HAM-D-17) scores for depression 
decreased from 19.8±4.4 (SD) to 13±5.35 (SD) after treatment (p=0.004).   
 
ELATED-2 was a double-blind, randomized, controlled study of 8 weeks of TLT 
(Omnilux New U - LED, 830 nm; 36.2 mW/cm2; up to 65.2 J/cm2; vs. sham for 16 
sessions). At endpoint, the mean HAMD-17 for depression decreased of 11.7±7.5 
points (TLT, n=9) vs. 5.3±7.0 points (sham, n=9) (p=0.04); the differences were even 
more pronounced in the completers sample as HAMD-17 decreased of 15.7±4.4 (TLT, 
n=6) points vs. 6.1±7.9 points (sham, n=7) (p=0.01).  
 
These findings suggest that TLT could be a novel intervention for subjects with MDD. 
Both the Laser study (ELATED-1) and the LED study (ELATED-2) were conducted by 
our group at Massachusetts General Hospital (MGH).  
 
NIR-flow (the present submission) will test for the first time the biological 
underpinning of TLT, by focusing on the effect of TLT on cerebral blood flow.   
 

 
  
 
                          Page 9 of 39                                                    C O N F I D E N T I A L                                           
.    
 
The LiteCure® The PhotoBioModulation-1000 (TPBM-1000) device to be used in this 
study does not require Food and Drug Administration (FDA) approval. In fact, the 
TPBM-1000device used in this study is considered a non-significant risk device 
because:  
 TPBM-1000 dose does not exceed the Maximum Permissible Exposure 
(MPE) for skin – Center for Devices and Radiological Health (CDRH),  
21CFR1040.10 and 21CFR1040.11;  
 TPBM-1000 is substantially equivalent to the OmniLux New U device (used 
in ELATED-2 at MGH), which the FDA has already categorized as a non-
significant risk device;    
 TPBM-1000 differs from the OmniLux New U  only in that it provides an 
additional site of irradiation – treatment is delivered to EEG sites FP1 and 
FP2 in addition to F3 and F4 on the forehead; 
 currently FDA-cleared LED devices are typically available over-the-counter 
and are considered safe to use without professional supervision.  
3.2 Overview of LiteCure Light Emitting Diode (LED )System 
LiteCure® TPBM-1000 is a new device with the flexibility to safely deliver either 
continuous light (c-TLT), pulsed light (p-TLT), or sham.  The TPBM-1000 device is 
configured for in-office use. 
 
TPBM-1000 delivers NIR light output at 830 +/- 30 nm.  The NIR output is invisible to 
the naked eye, thus, the device includes visual and/or audible indicators of light 
emission, and is designed to deliver light to EEG sites: Fp1, Fp2, F3 and F4, covering a 
total surface active treatment area, of 35.8 cm2[(11.52 cm2x2) + (6.38 cm2x2)]  with an 
average irradiance of 54.8 mW/cm2 and an average fluence of 65.8 J/cm2.  While the 
average fluence (energy density) is comparable to the Omnilux New U, the treatment 
area is significantly less for the TPBM-1000 (Omnilux New 28.7 cm2 x2, which results 
in less total energy delivered per session of 2.3 kJ (compared to up to 3.7 kJ for the 
Omnilux New U).   
 
TPBM-1000 devices will be fabricated to deliver either c-TLT, p-TLT, or Sham (s-
TLT). In simple terms, in either TLT mode the TPBM-1000 device delivers therapeutic 
NIR energy; in sham the device will not deliver any light energy. The apparent 
behavior (i.e. the performance/output of all visible and audible indicators) of the device 
in any of the three programmed treatment modalities will be identical, ensuring the 
study is blinded to the operator. In addition, because of the low average irradiances 
delivered in either c-TLT and p-TLT modes the subjects will not experience any 
heating sensation ensuring the study is single-blind. The device will shut off 
automatically if skin temperature above 41C were detected. 
 
The TPBM-1000 devices consists of a source of light, mounted on a cap, attached to a 
console that controls the light delivery modality: c-TLT, p-TLT, or Sham.  
 

 
  
 
                          Page 10 of 39                                                    C O N F I D E N T I A L                                           
.    
 
3.3 Overview of the DCS Device 
Separate hybrid FDNIRS-DCS instruments are described in detailed in the relative 
medical device brochure.   FDNIRS-DCS devices are approximately 23” (l) x 18” (w) 
x 9” (h). Technical specs are provided at: 
 iss.com/biomedical/instruments/metaox.html. The devices are limited by Federal (or 
United States) law to investigational use. The devices contain no conductive patient 
leads and no user serviceable parts. They have a highest rated current of 3 Amp and 
are designed for safety compliance with UL/IEC 60601-1 3rd Edition as a Class I, 
Type B device. Prior to investigational use, devices are tested for conformance with 
ground bond, earth leakage, and enclosure leakage requirements. The devices are not 
yet under Food and Drug Administration (FDA) investigational device exemption 
(IDE) regulations, and are considered non-significant risk. During the conduct of 
human studies the investigators should follow good clinical practice requirements and 
21 CFR 812 investigational device exemption regulations. 
 
The FDNIRS portion of the instrument consists of 8 radio- frequency (110 MHz) 
modulated laser diodes operating at 8 different wavelengths ranging from 670-830 
nm and 2 to 4 photomultiplier tube (PMT) detectors for heterodyne detection (similar 
to the Imagent from ISS, Inc).  
The DCS component of the instrument consists of a long-coherence length 852 nm 
laser, 4 to 8 low dark-count photon counting avalanche photodiodes, and a custom-
made correlator board. A short-pass filter in front of the FDNIRS PMTs, along with 
in-house designed software enables simultaneous FDNIRS-DCS measurements. 
Furthermore, to customize this instrument for use in humans, the MGH group has 
designed optical probes which host fiber optics. Each probe consists of 1 source and 
up to 8 detector fiber bundles terminated with 90 deg prisms providing multiple 
source-detector separations (ranging from 0.3 to up to 5 cm).   
The optical parameters for the FNIRS- DCS devices can be summarized as follows, 
irradiance 250-400 mW/cm2; area: 0.0962 cm2 and 10 min exposure (twice 10 min, 
before and after the session with TPBM-1000); fluence: 150-240 J/cm2. Total energy 
delivered in 10 min by the FNIRS-DCS devices: 14.43 – 23.08 J (this energy will be 
delivered twice before and after each TPBM-1000 sessions). The incremental 
exposure to NIR resulting from the FNIRS-DCS devices are minimal: in fact the NIR 
light is shed to a tiny area 1/350 of the area irradiated by TPBM-1000 and the energy 
shed in 10 min by the FNIRS-DCS are inferior to the total energy shed by the TPBM-
1000 by more than 2 orders of magnitude.   
 
3.1 Overview of the EEG Device 
 
The StatX24 (Advance Brain Monitoring) is an internally battery powered Type BF 
device intended for continuous use (16-17 hours). The StatX24 provides an integrated 
approach for wireless acquisition and recording of electroencephalographic (EEG), 
electrooculographic (EOG), and electrocardiographic (ECG) signals. The system 

 
  
 
                          Page 11 of 39                                                    C O N F I D E N T I A L                                           
.    
 
utilizes the patented Sensor Headset and patented EEG sensors, which record high 
quality EEG, obtained with less than five-minutes of set-up time and no scalp 
abrasion required. The wireless technology allows the user to be un-tethered and 
move around the home, healthcare facility, or clinical research environment while 
real time data is collected and displayed. 
The StatX24 acquires twenty channels of monopolar EEG recordings with a linked 
mastoid reference and optional channel for ECG, EOG, or EMG. The 1-10th 
channels, as well as the 21st channel, are differential channels. The 22nd-24th 
channels are referential channels. The StatX24 consists of: (1) StatX24 Headset with 
a Bluetooth (BT) Receiving Unit for bi-directional transmission of digitized 
physiological signals, (2) a Neoprene Strap, and (3) a Strip with EEG sensors sites in 
the standard 10-20 format + POz (Fz, Cz, Pz, F3, F4, C3, C4, P3, P4, O1, O2, T5, T3, 
F7, Fp1, Fp2, F8, T4, and T6 with Linked Mastoids). 
The Sensor Headset collects signals from the sensors placed on the patient and 
performs analog-to-digital conversion, encoding, formatting, and transmitting of all 
signals. The signals communicate using a 2.4 to 2.48 GHz radio transmitter. StatX24 
acquisition utilizes the bi-directional capabilities of the system to initiate scalp-
electrode impedance monitoring and monitors the battery capacity in the StatX24 
Headset. A BT Receiving Unit is used as the base unit affixed to the PC workstation. 
3.2 Overview of Rationale for Diffuse Correlation Spectroscopy Cognitive 
Tasks and Electroencephalogram 
Diffuse Correlation Spectroscopy 
From a cellular and molecular perspective, the beneficial effect of TLT (NIR) on 
brain metabolism is the primary putative mechanism for its therapeutic effects.  In 
experimental and animal models NIR (and red) light delivered noninvasively is 
absorbed by cytochrome c oxidase, and by stimulating the mitochondrial respiratory 
chain, leads to increased adenosine triphosphate (ATP) production 22. In a study on 
healthy subjects, TLT (NIR) improved cerebral oxygenation, supposedly through a 
mechanism of neurovascular coupling, which entails an increase in cerebral blood 
flow coupled with increased oxygen demands 12.  Similarly, a study in MDD subjects 
found that a single session of t-PBM led to a nearly significant increase in regional 
cerebral blood flow 2 ─ both studies relied on functional NIR spectroscopy (fNIRs) to 
assess, respectively, oxygenation and blood flow.  In a case report of moderate, 
chronic TBI, the improvement in brain perfusion subsequent to TLT was imaged with 
single positron emission computerized tomography (SPECT) 23. 
 
Cognitive Tasks 
The prefrontal cortex (PFC) and dorsal lateral prefrontal cortex (DLPFC) are 
uniquely situated to play a critical role in learning, executive functions, attention and 
inhibition. Studies have shown that DLPFC exhibits significant plasticity during the 
learning process, in which neuronal activity changes to dynamically encode and 
recode procedural memories once a sensory motor association has been made. The 
purpose of this study is to determine whether TLT can be used to change the speed at 
which subjects learn and perform in cognitive tasks.  TLT applied to prefrontal 

 
  
 
                          Page 12 of 39                                                    C O N F I D E N T I A L                                           
.    
 
regions of the brain implicated in visual-motor learning processes can enhance neural 
plasticity and improve behavioral performance. 
 
Electroencephalogram 
EEG recorded at the scalp offers the opportunity to access the synchronization of 
oscillatory frequencies (1Hz to 100Hz) from within and between cortical regions, 
revealing network changes that reflect abnormal connectivity. Synchronization within 
and between cortical regions is of interest in the field of TLT. In animal models of 
depression, TLT was found to regulate the EEG tracing in all wave frequency bands 
except for theta. Similar effects have been documented in dementia subjects who 
experienced normalization of delta and alpha waves power during the course of a 
pilot study with TLT treatment (Berman M & Hamblin M unpublished data).   
 
 
4. Investigational Plan 
4.1 Rationale 
The specific aims of this study are: 
1. To assess the change in cerebral blood flow induced by the Transcranial Continuous 
and Pulse Near-Infrared Light Therapy (c-TLT and p-TLT) in healthy subjects 
2. To correlate with cognitive performance the change in cerebral blood flow induced 
by the Transcranial Continuous and Pulse Near-Infrared Light Therapy in healthy 
subjects  
3. To correlate with brain electrical activity the change in cerebral blood flow induced 
by the Transcranial Continuous and Pulse Near-Infrared Light Therapy in healthy 
subjects 
4. To correlate with skin pigmentation the change in cerebral blood flow induced by 
the Transcranial Continuous and Pulse Near-Infrared Light Therapy in healthy 
subjects 
5. To assess the safety and tolerability of the Transcranial Continuous and Pulse Near-
Infrared Light Therapy in healthy subjects 
 
 
 
4.2 Study Design 
This study is a single-blind, sham-controlled, study on the use of Transcranial 
continuous light therapy (c-TLT) and pulse light therapy (p-TLT) as an enhancer of 
cerebral blood flow in healthy subjects.    
 
We expect that out of 30 subjects enrolled 10 subjects will be eligible to complete a 
sequence of three sessions of Transcranial Light Treatment (TLT) and sham.   
 

 
  
 
                          Page 13 of 39                                                    C O N F I D E N T I A L                                           
.    
 
All eligible subjects will receive two TLT sessions and one of sham (s-TLT) within 
the course of 5 weeks. While the sequence (c-TLT, s-TLT, p-TLT) will be the same 
for all subjects, the subjects will be blind to the specific mode of function used in 
each session.  
 
The study site (MGH) will have one Litecure device, which will recognize the 
subjects’ codes generated by the study statistician. Only subjects will be blind to the 
treatment group assignment. The Sham treatment consists of applying all the 
procedures for the delivery of the TLT, while actually delivering no light. The 
blinding of the sham will be effective because the radiation is not visible and because 
the active therapy (continuous or pulsed) will not produce any subject-discernable 
difference from sham, e.g., skin warming. The sham treatment will mimic the TLT 
procedure. The treatment will be bilateral and applied to the frontal areas with two 
application sites on the left side and two on the right side [left and right forehead 
centered on EEG sites on F3, F4, Fp1, Fp2]. Energy is administered with a radiation 
wavelength of 830 nm. The duration of irradiation is 20 minutes at each application 
site (the 4 sites are irradiated at the same time which is equivalent to 20 minutes of 
total time). The entire sessions is estimated to last about 1.5 hours. The additional 
time is needed to have the subject complete a urine toxology and self-report forms, to 
prepare the subject, to place the necessary protections (e.g. goggles), to inspect the 
subject’s skin, to complete DCS before and after TLT, to set the TLT devices, and to 
give the subject time to rest after the irradiation. TLT treatment will only be 
administered by licensed physicians (i.e. MDs) who are on study staff. All staff who 
deliver treatment must pass training that is approved by the MGH Laser Safety 
Committee. The treatment will follow these specifications: wavelength 830 nm; 
average irradiance 54.8 mW/cm2; average fluence of 65.8 J/cm2 (consistently with  
energy density used in the ELATED-2 study at MGH with the device Omnilux New 
U). 
4.3 Study Population 
 
Healthy subjects who meet all inclusion criteria, have none of the exclusion criteria, 
and are willing to undergo the Informed Consent process will be eligible to 
participate in this study.  
4.3.1 Population Description 
The subject population for this study will consist of healthy subjects between 
the ages of 18 and 70, of any ethnic background. We expect to enroll 30 
subjects in order to have 10 who are eligible and complete the study. The 
subject will meet all the inclusion criteria, have none of the exclusion criteria, 
and provide their written Informed Consent to participate in this clinical study.  
In the event that a subject’s ability to comprehend and communicate is 
compromised (per the assessment of the Investigator), local regulations 
pertaining to Informed Consent signatures should be followed. 
 

 
  
 
                          Page 14 of 39                                                    C O N F I D E N T I A L                                           
.    
 
Subjects will be recruited, via screenings, advertisements, and postings. Study 
participants will also be recruited via internet-based advertisements, e.g., 
Craigslist, Clinical Trials @ Partners, the study site’s websites, RSVP for 
Health (which will also generate listserv emails to Partners employees, 
regarding research studies in need of volunteers), etc.   
 
Participants who contact the respective study site staff in response to any of 
the aforementioned advertisements will complete phone screening questions 
to determine preliminary eligibility for the study.  
4.3.2 Use in Female Subjects of Childbearing Potential 
Female subjects of childbearing potential must consent (without any element 
of coercion) to use a double-barrier method for birth control (e.g. condoms 
with spermicide) if sexually active.  A pregnancy test will be performed at 
the Screening Visit.  
 
4.3.3 Criteria for Enrollment – listed on pages 5-6 
4.4 Subject Numbering 
Subjects enrolled in the NIR-flow study will be identified by subject numbers 101 
through 130 (to account for screen failures and drop-out before TLT sessions). 
4.5 Sample Size 
The NIR-flow study is expected to enroll 30 subjects, with 10 subjects who will be 
eligible and have analyzable data (at least 1 session post randomization).   
 
4.6 Subject Confidentiality 
Any information and data provided to Litecure Inc. or its designees in reference to 
any subject’s participation in this investigation will be considered confidential. The 
Investigator will need to ensure that all subjects’ anonymity will be maintained on all 
documentation submitted to Litecure Inc by completely redacting (eliminating or 
“blacking out”) each subject's name and/or other identifying information. The 
identifying information will be replaced with the subject's study number and initials. 
The investigational site is not to provide to Litecure Inc information such as subject’s 
telephone numbers, home address, personal identification numbers such as passport 
numbers, etc. Care must be taken by site research personnel when communicating 
with representatives from Litecure Inc in the form of telephone or electronic 
correspondence in not providing information that may disclose a subject’s identity. 
 
Documents associated with the study that are not intended to be submitted to Litecure 
Inc (e.g., signed Informed Consent Forms) must be kept in strict confidence by the 
Investigator.  Only study site personnel, authorized Litecure Inc personnel, and 
regulatory authority inspectors will have access to these confidential files.  

 
  
 
                          Page 15 of 39                                                    C O N F I D E N T I A L                                           
.    
 
4.7 Study Visits 
Study visits occur at MGH Center for Anxiety and Traumatic Stress Disorder 
(CATSD) in Boston.  
 
The study involves screening, 3 TLT session paired with Diffuse Correlation 
Spectroscopy (DCS), and one follow-up visit.  
 
Screening Visit (Week 1)  
The screening visit is expected to take about 2 hours. The Principal Investigator (PI) 
at the site or a Sub-investigator will assess subjects against the inclusion and 
exclusion criteria.  If a subject appears to qualify and agrees to participate in the 
study, then the subject will undergo the informed consent process prior to initiation of 
any study-specific activities. Screening Case Report Forms (CRFs) will be completed 
and subjects who qualify to participate in the study will proceed to the first Treatment 
Visit 1. If the subject does not meet entry criteria, the Investigator will document the 
subject’s ineligibility for study participation and complete the appropriate CRFs.   
 
TLT-DCS Visits – Visits 2-4 (Weeks 2-4)  
There is one TLT session per week for 3 weeks. TLT-DCS Visit 2-4 will consist of 
all subjects receiving TLT, with DCS before and after TLT. Two groups of 
assessments will be conducted: 1. Before the TLT session to reflect the symptoms 
endorsed in the past week: Anxiety Symptoms Questionnaire (ASQ), Symptoms of 
Depression Questionnaire SDQ, Quality of Life in Neurological Disorders (Neuro-
QoL), Sexual Functioning Inventory (SFI), Systemic Assessment for Treatment 
Emergent Events (SAFTEE), and 2. After the TLT session to reflect the immediate 
changes associated with the session: Positive and Negative Symptoms Questionnaire 
(PANAS), TLT Self-Report Questionnaire (TSRQ), Perceptions of Blinding 
Questionnaire (PBQ). Cognitive tasks will occur right after the TLT session.  A 
detailed description of the study visits can be found below. EEG recording will 
immediately precede and follow the DCS-TLT-DCS triad and will in part overlap 
with the cognitive task.  
 
Follow-Up Visit – Visit 5 
All subjects will undergo assessments including ASQ, SDQ, Neuro-QoL, SFI, 
SAFTEE, PANAS and cognitive tasks at the follow-up visit (Week 5).  
 
4.8 Detailed Study Visits 
4.8.1 Screening Visit – Visits 1 (Week 1) 
Purpose 
The purpose of the Screening Visit will be to determine the eligibility of subjects 
for participation in the NIR-flow study. 
 
 

 
  
 
                          Page 16 of 39                                                    C O N F I D E N T I A L                                           
.    
 
Procedures 
The screening visit will occur at the MGH Center for Anxiety and Traumatic Stress 
Disorders (CATSD) in Boston. At this visit subjects will be consented for the study, 
will receive a psychiatric evaluation, and will be screened for presence of 
psychiatric disorders using the Structured Clinical Interview for Diagnostic 
Statistical Manual-IV (SCID-IV).  The SDQ, ASQ and PANAS will be used to 
assess for subthreshold symptoms. The SFI and NEURO-QoL will be used to assess 
sexual and cognitive functioning.  
 
Routine laboratory tests (Urine Tox Screen and Urine Pregnancy Test) and 
complete physical exam including vitals and weight will be performed at screen; the 
tests will be processed by a laboratory at MGH.  
 
Once all tests and procedures become available for review, the Investigator will 
then proceed to closely re-evaluate the subject for all the inclusion and exclusion 
criteria. 
4.8.2 TLT-DCS Visits – Visits 2-4 (Weeks 2-4)                                                                              
The TLT sessions will occur once weekly for three weeks at the MGH Martinos 
Center for Biomedical Imaging in Charlestown. Subjects missing one session will 
be allowed to still receive their TLT session, by shifting the overall study schedule 
of one week. The study PI will be an active participant in the delivery of the TLT. 
The PI and other collaborators from MGH CATSD will assist with the delivery of 
TLT. The device TPBM-1000 for the administration of the TLT will be provided by 
Litecure Inc. Litecure will provide training of MGH CATSD staff in the use of the 
device and will provide training certificates at the completion of the training.   
 
The overall study visit is estimated to last about 1.5 hours. The TLT session is 
estimated to last about 50 min – 5 minutes of EEG before DCS, 10 minutes of DCS 
before TLT, 20 minutes of TLT, 10 minutes of DCS after TLT and 5 minutes of 
EEG after DCS. EEG recording will extend to cover at least part of the time 
devoted to the cognitive tasks. Study sessions will take place in an office dedicated 
to TLT treatment at MGH Martinos Center for Biomedical Imagining in 
Charlestown. Only the subject and the staff administering the TLT will be allowed 
in during the session. The subject will comfortably lie down on an exam bed or sit 
on a recliner. The sites of application of TLT (left and right forehead) will be 
inspected for any possible skin lesions (e.g. laceration or signs of inflammation) 
which would contraindicate the treatment. The study staff administering the TPBM 
device at the TLT-DCS sessions will appropriately remove any reflective objects to 
fall into or obstruct the path of the light energy produced by this device. Both the 
subject receiving the TLT and the study staff will wear protective eyewear in the 
form of goggles.  During treatment sessions the office will be kept completely dark 
and a warning sign will hang at the office door. The staff will be provided training 
on basic safety procedures relative to the use of the device.  
 

 
  
 
                          Page 17 of 39                                                    C O N F I D E N T I A L                                           
.    
 
The trained staff administering the TLT will use utmost care to never shine the light 
in or near the eyes of the subject safety goggles will be selected based on the TLT 
biophysical properties, in accordance to the Occupational Safety and Health 
Administration (OSHA) guidelines. The delivery of the TLT is expected to last 20 
min total (simultaneous application on the left and right forehead). The subject will 
be asked to rest for five minutes after the delivery of TLT. The skin at the sites of 
the application will be inspected again prior to dismissing the subject. A log with 
the dates of treatment delivery per each study subject will be kept at MGH CATSD 
(Boston).  
 
During TLT-DCS Visit 2-4, subjects will receive TLT-c (Visit 2), TLT-s (Visit 3) 
and TLT-p (Visit 4).   
 
The following tasks will be completed prior to every TLT session:  
 
 Utox 
 Concomitant medications form 
 ADVERSE EVENTS FORM 
 ASQ (self-rated) 
 SDQ (self-rated) 
 NEURO-QoL (self-rated) 
 SFI (self-rated) 
 SAFTEE (self-rated) 
 PANAS (self-rated) 
 EEG- Recording 
 DCS – Spectroscopy 
 
The following tasks will be completed after every TLT session:  
 
 DCS – Spectroscopy  
 EEG- Recording  
 Cognitive tests 
 PANAS (self-rated) 
 TSRQ (self-rated) 
 PBQ (self-rated) 
 
Subjects shall be instructed to contact the investigator or a member of his staff at 
any time between visits concerning adverse events or worsening of symptoms.   
4.8.3 Follow-up Visit – Visit 5, (Week 5) 
After treatment completion, subjects will continue with one more clinical study visit to 
measure the long-term pro-cognitive effect of TLT. The follow-up visit will typically 
occur at the MGH Center for Anxiety and Traumatic Stress Disorders (CATSD) in 
Boston but could be performed at the Martinos Center for Biomedical Imaging in 
Charlestown if preferable to the subject. 

 
  
 
                          Page 18 of 39                                                    C O N F I D E N T I A L                                           
.    
 
 
 
At Visit 5, the following will be performed: 
 
 
 Concomitant medications form 
 ADVERSE EVENTS FORM 
 ASQ (self-rated) 
 SDQ (self-rated) 
 NEURO-QoL (self-rated) 
 SFI (self-rated) 
 SAFTEE (self-rated) 
 PANAS (self-rated) 
 Cognitive tests 
4.8.4 Termination 
We will encourage subjects to complete the follow-up visit in the study regardless of 
discontinuation of the TLT sessions.   
4.9 Scales & Forms 
We will offer clinical interviews and study forms in English. All scales will explore 
the past 7 days except for SCID (lifetime), PANAS (past 5 min), TSRQ and PBQ 
(both past session). Below are brief descriptions of the forms that will be utilized in 
this study.  
 Structured Clinical Interview for DSM-IV (SCID-I/P) – The SCID-I/P, 
administered by the clinician, proceeds by modules to diagnose the different 
psychiatric disorders. Questions here are asked exactly as written, and each is 
based on the individual criteria from DSM-IV. Answers are generally rated on a 
scale of 1-3 (1 = doubtful, 2 = probable, 3 = definite), and based on the number of 
positive answers, a diagnosis is determined. The entire SCID-I/P is administered 
at screening as per Table 1.   
 Symptoms of Depression Questionnaire (SDQ) – This is a comprehensive 
measure of depression that includes the assessment of symptoms in the anxiety–
depression spectrum. It assesses irritability, anger attacks, and anxiety symptoms 
together with the commonly considered symptoms of depression. Analysis of the 
factor structure of the SDQ identified 5 subscales, including one in the anxiety–
depression spectrum, with adequate internal consistency and concurrent validity.  
 Anxiety Symptoms Questionnaire (ASQ) – This, is a 17-item self-report 
questionnaire measuring the frequency and intensity of 17 symptoms of anxiety, 
including nervousness, worrying, irritability, trouble relaxing, insomnia, lack of 
energy, difficulty concentrating, somatic symptoms, and impairment in 
functioning due to anxiety.  
 Systematic Assessment for Treatment Emergent Events (SAFTEE-SI) – The 
SAFTEE is a commonly used instrument originally developed by NIMH and 
adapted into a self-report instrument. The version of the scale that we plan to use 

 
  
 
                          Page 19 of 39                                                    C O N F I D E N T I A L                                           
.    
 
is the same used by the multi-center, NIMH-sponsored CO-MED trial, and it 
examines in a systematic fashion all possible treatment-emergent side effects and 
probes specific adverse symptoms, including suicidal thoughts and behaviors, and 
self-injurious behavior.  
 Positive and Negative Symptoms Questionnaire (PANAS) – The 20-item 
PANAS, comprises two mood scales, one measuring positive affect and the other 
measuring negative affect. Each item is rated on a 5-point scale ranging from 1 = 
very slightly or not at all to 5 = extremely to indicate the extent to which the 
respondent has felt this way in the indicated time frame. The authors have used 
the scale to measure affect at this moment, today, the past few days, the past 
week, the past few weeks, the past year, and generally (on average). 
 Adverse Events Form- the Adverse Events Form captures any adverse event 
(serious or otherwise) specifically related to the application of the TLT. This 
form will help to determine if the side effects of the TLT are too great for a 
participant to continue in the study. 
 Quality of Life in Neurological Disorders (Neuro-QoL Item Bank v2.0 – 
cognition function– short form) - The cognitive section of the Neuro-QoL is an 
8 item self-rated measure of both executive function and general concerns. It 
measures perceived difficulties in cognitive abilities (e.g., memory, attention, and 
decision making) or in the application of such abilities to everyday tasks (e.g., 
planning, organizing, calculating, remembering and learning). 
 Massachusetts General Hospital Sexual Functioning Inventory (MGH-SFI) -
This is a subject-rated self-report outcome measure that quantifies sexual 
dysfunction into 5 functional domains (“interest in sex,” “sexual arousal,” “ability 
to achieve orgasm,” “ability to maintain erection” [males only], and “sexual 
satisfaction”).  
  The TLT Self-Report Questionnaire (TSRQ) – An open-ended questionnaire 
focusing on potential inconveniences and discomforts from the TLT.  
 Concomitant medications form – This form will be completed at every TLT 
study visit, including the screening visit, as a safety-monitoring tool.  
 The Perceptions of Blinding Questionnaire (PBQ): the PBQ is a self-report 
questionnaire to determine the degree to which the participant believes s/he is 
receiving the treatment or the sham.  
4.10 Cognitive tests 
n-Back Task: The subject is presented with a sequence of stimuli, and the task consists 
of indicating when the current stimulus matches the one from n steps earlier in the 
sequence. The load factor n can be adjusted to make the task more or less difficult. 
The n-back task captures the active part of working memory. When n equals 2 or more, 
it is not enough to simply keep a representation of recently presented items in mind; the 
working memory buffer also needs to be updated continuously to keep track of what the 
current stimulus must be compared to. To accomplish this task, the subject needs to both 
maintain and manipulate information in working memory.  

 
  
 
                          Page 20 of 39                                                    C O N F I D E N T I A L                                           
.    
 
4.11 EEG recording 
The participant will be fitted with an EEG sensor headset and ECG leads to start their 
resting state data acquisition. For resting state acquisition with eyes open, the participant 
will be asked to relax, sit still and focus on a cross on the computer for 5 minutes while 
EEG data is acquired. The participant will be instructed to try to limit blinking and any face 
movement including clenching their jaw. Several acquisitions and/or longer acquisitions 
may be necessary if the participant is particularly restless. The participant will then be 
asked to relax and close their eyes for 5 minutes during the eyes closed resting state 
acquisition. Multiple and/or longer eyes closed acquisitions may be necessary if the 
participant has difficulty or is unable to sit still for 5 minutes. Any issues, including 
inability to remain still for the acquisition, should be noted in the participant's file. 
 
 
Data Collection Forms 
All study subjects will be blind to TLT assignment (single-blind). A number of 
procedures are in place to assure data integrity and protocol adherence. We will use 
Research Electronic Data Capture to support direct data entry by subjects and study staff. 
REDCap is a free, secure, HIPAA compliant web-based application hosted by the Partners 
HealthCare Research Computing, Enterprise Research Infrastructure & Services (ERIS) 
group. Web-based surveys rely on a study-specific data dictionary defined by members of 
the research team with planning assistance from Harvard Catalyst/The Harvard Clinical and 
Translational Science Center EDC Support Staff. These support staff will then oversee the 
automated export of study data from REDCap to a relational study database in Microsoft 
Access 2000, allowing for systematic data querying and checking.  
Self-report measures will be completed by participants on a computer, directly into 
REDCap, thus minimizing errors due to data entry.  For clinician-administered measures, 
all clinicians and independent evaluators (IEs) will enter responses directly in REDCap.  
 
To minimize missing data for self-report forms, we will program missed question 
warnings in REDCap that will alert participants in real-time if they inadvertently skip a 
question.  Participants may then go back and answer any missed questions, or, if they 
intentionally skipped questions, they may ignore the warning message and continue answer 
the remaining questions.  We will also program real-time range checks in REDCap that 
generate error messages if a value outside the acceptable range is entered for a given field.  
To ensure confidentiality, data will be identified in the database only by subject number, 
visit number, and date of visit. By recording the study data in this manner, the information 
can be considered ‘de-identified’ and therefore compliant with the Standards for Privacy of 
Individually Identifiable Health Information (“Privacy Rule”) of the Health Insurance 
Portability Act of 1996 (HIPAA). Any data that is transmitted electronically will be fully 
encrypted and password protected. Subjects’ names will not be entered into the database; 
each will be uniquely identified only by an ID number. Consent forms, any hard copy PHI, 
and any study measures that are completed on paper will be kept and filed in locked office 
cabinets.  

 
  
 
                          Page 21 of 39                                                    C O N F I D E N T I A L                                           
.    
 
4.12     Data Management 
The principal investigator, Dr. Cassano is ultimately responsible for the quality of 
the data collection and overall conduct of the study, and directly supervise the study 
coordinators and data management staff at MGH. Data management will include clinician 
and subject rated assessments (see assessments), screening data, fidelity data, visit 
adherence data and rater reliability data and safety reporting, including DCS and cognitive 
data. Weekly reports from REDCap will monitor subject enrollment, completion, attrition, 
and individual subject progress as well as the completion of critical assessments. 
Additional reports will be done as needed to monitor baseline characteristics, protocol 
adherence, and other issues of interest.  
 
All measures that are completed on paper will be entered by the RA into REDCap, and 
a two pass verification system will be used to minimize any data entry errors. All records in 
REDCap have a form completion status that may be Incomplete (appears as a red circle in 
Record Status Dashboard), Unverified (yellow circle), or Verified. The RA initially 
entering data will save each record entered as Unverified (at which point it will appear as a 
yellow circle in the Record Status Dashboard). A second RA at each site will then go into 
each unverified record, compare each entered value against the paper source document, 
make any corrections, and then re-save the record as Verified (at which point it will appear 
as a green circle in the Record Status Dashboard). The color coding system built into 
REDCap readily allows for identification of unverified records. 
 
Coded data may be sent to collaborators within and outside of Partners for research 
purposes. Coded sample/data may be shared with for profit companies that are working 
with Partners researchers, such as Litecure, LLC or Advanced Brain Monitoring, Inc (the 
manufacturer of the EEG device used this in study). Samples/data will not be sold to 
anyone for profit.  Data may also be shared with other Partners IRB approved protocols 
supervised by the principle investigator of this study. 
 
4.13     Training 
Trained study clinicians will administer diagnostic assessments and rating scales. 
All raters will be experienced clinicians who will have undergone specific training to 
criteria in the use of the study measures.  
4.14      Blinding of the Study 
Since this is a single-blind, sham-controlled study, measures have been taken to 
assure that the integrity of the blinding of the treatment modalities remains intact. The two 
means of assuring that the treatment modalities will not be disclosed to subjects include 
detectors sensitive to heating of the skin (which will switch off the device to prevent 
overheating) and the introduction of a rating scale for subjects to assess their best guess on 
treatment assignment. 
 

 
  
 
                          Page 22 of 39                                                    C O N F I D E N T I A L                                           
.    
 
The performance/output of all visible and audible indicators is identical for all three 
treatment modalities, (i.e. other than the emission of invisible light radiation, the behavior 
of the system is identical for all).  
4.15      Compensation 
 
Subjects will be compensated $50 per screen (only if deemed eligible) and $100 per 
TLT-DCS visit, and $50 per follow-up visit; for a total of $400, if all five visits are 
completed. Compensation will be provided after Weeks 2 ($150), 3 ($100), 4 ($100), and 5 
($50), with the correspondent amount subtracted for each visit missed. Any research 
participant parking in one of the three Mass General Hospital garages will receive a 
parking voucher.  
 
4.16      Accountability/Investigational Product Control 
 
U.S. federal law and ICH Guideline E6 § 5.14 requires that all investigational 
medical devices be strictly controlled. All study devices must be kept in a secured area at 
the clinical sites in compliance with all applicable FDA (U.S. sites) regulations.  
 
 
The Principal Investigators or designated study site personnel who verify the receipt 
of the devices/device accessories must complete the Device/Accessories Acknowledgment 
Form and fax a copy to Litecure Inc.  Device Accountability and Acknowledgment Logs 
will be maintained at each study site.  These logs will list all equipment received, the 
receiving date, the serial number of each device.  Study site personnel will initial the log 
each time the device is used. TPBM-1000 use will also be recorded on the appropriate 
CRF.   
 
 
Malfunctioning devices and device accessories, including all components, will be 
returned to Litecure Inc. for investigation, at Litecure expense.   
4.17      Clinical Adverse Events 
 
4.17.1 Overview and Definitions  
All adverse events will be recorded from the time of Informed Consent 
through study completion, or termination.  The Adverse Event CRF must be 
completed and submitted to the IRB, FDA, and Litecure, as required.  Regulations 
for adverse event handling and reporting contained in the FDA and ICH Guidelines 
will be adhered to. 
 
Consideration of Adverse Events will hereafter consist of Adverse Events, 
Serious Adverse Events, and Adverse Device Effects, including Anticipated 
Adverse Device Effects and Unanticipated Adverse Device Effects.  
 
 Adverse Event is defined as any untoward/undesirable clinical occurrence in 
a clinical investigation of a subject using a device and/or product and which 

 
  
 
                          Page 23 of 39                                                    C O N F I D E N T I A L                                           
.    
 
does not necessarily have a causal relationship with this treatment. An 
Adverse Event can therefore be any unfavorable and/or unintended sign, 
symptom, or disease temporally associated with the use of a device product, 
whether or not considered related to the device product. Only abnormal 
laboratory values that are deemed clinically significant by the investigator 
will be classified as adverse events. 
 Serious Adverse Event is defined as any untoward/undesirable adverse 
experience that results in any of the following outcomes:  1) death; 2) a life-
threatening adverse experience; 3) inpatient hospitalization or prolongation 
of existing hospitalization; 4) a permanent/persistent or significant 
disability/incapacity or a congenital anomaly/birth defect; 5) important 
medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered a serious adverse experience when, based 
upon appropriate medical judgment, they may jeopardize the subject and 
may require medical or surgical intervention to prevent one of the outcomes 
listed in this definition.  
 Anticipated Adverse Device Effect is defined as any adverse effect related to 
the device or procedure, which is identified in the protocol. 
 Unanticipated Adverse Device Effects is defined as any serious adverse 
effect on health or safety or any life-threatening problem or death caused by, 
or associated with, a device, if that effect, problem, or death was not 
previously identified in nature, severity, or degree of incidence in the 
investigational plan or application (including a supplementary plan or 
application), or any other unanticipated serious problem associated with a 
device that relates to the rights, safety, or welfare of subjects.  
 
4.17.2 Safety Monitoring 
The study subjects will undergo frequent clinical evaluations including 
mood and anxiety scores, concomitant medications, adverse events, and serious 
adverse events and unexpected device events will be recorded from study entry 
through completion. 
 
If skin erythema is present, treatment will be suspended. Subjects will be 
instructed to contact the study site principal investigator or a member of his staff at 
any time between visits concerning adverse events or worsening of subthreshold 
symptoms. If at any study visit the subjects’ clinical condition is significantly 
worsened from baseline, the subject will be referred to appropriate level of care. 
(see 4.8.4 Termination).    
 
Recording of physiological data with the EEG system poses no known 
health risk. Participants may experience mild discomfort due to the pressure exerted 
by the sensor headset; to minimize this risk, caps are adjustable and sensor strips are 
available in a variety of sizes. Discomfort may also arise from a mild allergic 
response to extended exposure to the conductive gel used in the EEG sensors; to 
mitigate this risk, alcohol wipes will be available to remove the gel immediately to 
reduce any allergic reactions. 

 
  
 
                          Page 24 of 39                                                    C O N F I D E N T I A L                                           
.    
 
 
 
The site Principal Investigator (Dr. Cassano) will have monthly conference 
calls or meetings (which will include study RAs); during these conference calls he 
will discuss all Adverse Event reports to identify any safety concern, based on such 
concerns he will be able to decide temporary discontinuation of study enrollment, 
modifications of the study protocol, or to terminate the study.   
 
Research assistants responsible for data collection and storage will be aware 
of and comply with all regulatory requirements related to adverse events.  In the 
event that a subject becomes ill or is injured as a direct result of study participation, 
medical care will be made available.  All adverse events (and device events) will be 
followed to resolution and reported to the MGH IRB as serious in the event that 1. 
they are unanticipated and possibly related to the study (same reporting as SAE) or 
2. if they meet any one of the following criteria: Any fatal event, immediately life-
threatening event, permanently or substantially disabling event, event requiring or 
prolonging inpatient hospitalization, or any congenital anomaly. This also includes 
any event that a study investigator judges to impose a significant hazard, 
contraindication, side effect, or precaution.   
 
Expedited review will occur for all events meeting the FDA definition of 
Serious Adverse Events (SAEs) – i.e., any fatal event, immediately life-threatening 
event, permanently or substantially disabling event, event requiring or prolonging 
inpatient hospitalization, or any congenital anomaly. This also includes any event 
that a study investigator judges to impose a significant hazard, contraindication, 
side effect, or precaution.  Notification by e-mail of all related study forms shall be 
made to the IRB within 2 days of the occurrence of any SAE.  Information will be 
reviewed and a determination made of whether there was any possible relevance to 
the study TLT exposure.  Additional reporting to local IRBs will be done within 5 
business days/7 working days of the date the investigator first becomes aware of the 
problem.  
            4.17.3 Reporting Procedures for All Adverse Events 
After review with the subject by the study site personnel, all Adverse Events 
occurring during the study, whether or not attributed to the TPBM-1000 device or 
TLT procedure, observed by the Investigator or reported by the subject, will be 
documented in the subject’s source document and on the appropriate CRF pages.  
The following attributes must be assigned:  
 
1. Description of event 
2. Date of onset 
3. Date of resolution (if applicable) 
4. Seriousness  
5. Relationship to the study device and/or procedure(s) 
6. Intensity 
7. Action(s) taken  

 
  
 
                          Page 25 of 39                                                    C O N F I D E N T I A L                                           
.    
 
8. Outcome(s) 
 
Intensity is defined as a measure of the severity of a reaction, effect or experience. 
The measurement(s) are described as mild, moderate or severe. The event itself, 
however, may be of relative minor medical significance. 
 
The intensity of Adverse Events is assessed as mild, moderate or severe according 
to the following index scale: 
 
Mild 
The Adverse Event is transient, requires no treatment, and does not interfere 
with the subject’s daily activity. 
Moderate 
The Adverse Event introduces a low level of inconvenience or concern to 
the subject and may interfere with daily activities, but is usually ameliorated 
by simple therapeutic measures. 
Severe 
The Adverse Event interrupts the subject’s usual daily activity and requires 
systematic therapy or other treatment. 
 
If the Adverse Event is of such intensity in the Investigator’s judgment that it 
warrants withdrawal from the study, the subject should be withdrawn from 
treatment. The subject should be given appropriate care under medical supervision 
until symptoms resolve.   
 
The relationship of an Adverse Event to the study device or procedure will be 
graded as follows: 
 
None 
The Adverse Event is not associated with the study device use. 
Remote 
The temporal association is such that the study device is not likely to have 
had an association with the observed Adverse Event. 
Possible 
This causal relationship is assigned when the Adverse Event:  
a) Follows a reasonable temporal sequence from device use, but  
b) Could have been produced by the subject’s clinical state or other modes 
of therapy administered to the subject. 
Probable 
This causal relationship is assigned when the Adverse Event:  
a) Follows a reasonable temporal sequence from device use;  
b) Abates upon discontinuation of the treatment;  
c) Cannot be reasonably explained by known characteristics of the 
subject’s clinical state.  
Definite 
This causal relationship is assigned when the Adverse Event:  

 
  
 
                          Page 26 of 39                                                    C O N F I D E N T I A L                                           
.    
 
a) Follows a reasonable temporal sequence from device use;  
b) Abates upon discontinuation of the treatment; and  
c) Is confirmed by the reappearance of the Adverse Event on repeat 
exposure. 
 
*For purposes of reporting Unanticipated Adverse Events, “None” and “Remote” 
will be considered as having no association with the device or treatment procedure. 
 
4.17.4 Serious Adverse Events 
All Serious Adverse Events must be reported to the overseeing Institutional Review 
Board (IRB), FDA, and Litecure Inc as required.  
 
If the Investigators are notified by Litecure Inc or its designee of any Serious 
Adverse Events that are considered to be Unanticipated Adverse Device Effects, the 
Investigators must notify his/her own IRB/EC as required. 
 
4.17.5 Deaths 
Deaths which must be reported to Litecure Inc include all deaths while participating 
in the study. 
 
4.17.6 Withdrawals for Adverse Events 
All Adverse Events which result in the subject’s withdrawal from the study must be 
reported immediately by telephone to Litecure Inc. 
  
The Investigator may be asked to provide detailed follow-up information.  The 
Investigator will determine the reportability of the event on a case-by-case basis, 
and will report to the appropriate regulatory authorities evaluating the study device 
as necessary.   
4.18 Measures to Assure Subject’s Safety 
The Investigator will be responsible for monitoring the safety of subjects who enter 
this study and for alerting Litecure Inc of any study-related event that seems unusual and/or 
unanticipated for his/her site.  
 
The Investigator will be responsible for the appropriate medical care of the subjects 
during the study in connection with protocol procedures for his/her site. 
 
The Investigator will remain responsible for providing any appropriate health care 
options after a subject’s completion or discontinuation from the study due to adverse events. 

 
  
 
                          Page 27 of 39                                                    C O N F I D E N T I A L                                           
.    
 
4.19 Subject Disposition Criteria 
4.19.1 Withdrawal from the Study 
Each subject and the Investigator reserve the right at any time to terminate a 
subject’s participation in the clinical investigation.   
 
Possible reasons for withdrawal or removal from the study may include: 
 
1. The subject voluntarily withdraws consent. 
2. The subject was not eligible based on the study inclusion and exclusion 
criteria. 
3. The subject develops an Adverse Event that would not allow continuation in 
the study. 
4. The subject has an Adverse Event which in the opinion of the Investigator 
warrants withdrawal from the study. Litecure Inc must be notified within 
two business days. 
5. A decision is made by the subject and/or Investigator that the subject should 
be withdrawn from the study. 
6. Subject death  
7. Positive pregnancy test during the eligibility procedures  
 
Treatment will end for participants who experience temporary pain or discomfort 
during a treatment session. A research assistant or the study doctor will follow-up 
within a 24-hour period, after the discontinued treatment, with the patient who 
experienced discomfort. The subject may continue with the study if the acute pain 
stopped. If the subject experiences discomfort after 24-hours the aforementioned 
treatment discontinuation process may begin.     
When a subject withdraws or is removed from the study, the following will be 
performed, if feasible, at the study termination (exit) visit: 
 
 Concomitant medications form 
 ADVERSE EVENTS FORM 
 ASQ (self-rated) 
 SDQ (self-rated) 
 NEURO-QoL (self-rated) 
 SFI (self-rated) 
 SAFTEE (self-rated) 
 PANAS (self-rated) 
 Cognitive tests 
 
For all subjects who withdraw from the study prematurely, the date, and reason for 
withdrawal will be documented. 

 
  
 
                          Page 28 of 39                                                    C O N F I D E N T I A L                                           
.    
 
4.19.2 Lost to Follow-up 
If the site is unable to contact a subject or if the subject fails to appear for a visit, 
three documented phone calls should be made. The site’s unsuccessful attempts to 
contact the subject and the subject’s failure to contact the site will result in the 
subject being withdrawn from the study. The subject will be considered Lost to 
Follow-up.  
 
4.20 Risk and Benefits Overview 
4.20.1 Potential Adverse Events 
Risks to the subject may include but are not limited to the following: 
The TPBM-1000 emits light with a longer wavelength than the human eye can see. 
The TPBM-1000 output is less than the maximum permissible exposure (MPE). 
The staff will be provided training on basic safety procedures relative to the use of 
the device. The staff administering the TLT will be careful not to operate the 
TPBM-1000 device unless it is in direct contact with the subject’s skin. Both the 
subject receiving the TLT and the study clinician present in the TLT delivery room 
will wear protective eyewear in the form of goggles or eye pads.  The goggles 
provided with the TPBM-1000 are in direct contact with the area surrounding the 
subject’s eye.  Since they are reusable they should not be shared between subjects.  
The eye pads are disposable and should be discarded after use.  The eye pads use an 
adhesive to adhere to the subject’s eyelids so there is potential for an allergic 
reaction. 
 
Failure of the TPBM-1000, resulting in the cessation of investigative therapy can 
cause: 
1. No adverse event to our knowledge 
2. Unforeseeable adverse events 
 
Delivery of the infrared energy to an inappropriate site, such as directly over the 
open eye, is not recommended and could pose a risk to the subject. 
 
Application of the TPBM-1000 may result in mild thermal sensation of 
warmth during the use.  The temperature of the skin is well below the level for 
thermal damage. 
 
Based on human clinical trial experience to date, each adverse event listed 
below has been reported with TLT: 
1. 
Application Site Erythema 
2. 
Application Site Pain 
3. 
Application Site Discomfort 
4. 
Application Site Warmth 
5. 
Application Site Reaction 
6. 
Headache 

 
  
 
                          Page 29 of 39                                                    C O N F I D E N T I A L                                           
.    
 
 
Additional potential side effects of TLT, documented at MGH in prior trials, 
include:  
 
- 
Seeing vivid colors, having abnormal taste 
- 
Feeling “out-of-body” experiences 
- 
Insomnia, restless sleep, erratic sleep, early morning awakenings 
- 
Vivid dreams 
- 
Irritability 
- 
Word finding difficulties 
- 
Abdominal bloating 
 
Other potential risks are described below: 
 
Risk of Worsening of lingering Depression of Anxiety and Manic Switch:   
We will minimize this risk by selecting only healthy subjects.  We will also 
discontinue any subject who develops active suicidal ideation during the course of 
the study; we will then arrange for appropriate levels of care and standard 
antidepressant treatment (see 4.8.4 Termination).  Manic switches are possible 
adverse events and will be closely monitored during treatment as well.  Subjects 
who develop mania or hypomania will be discontinued and provided appropriate 
level of care.  
 
Answering detailed questionnaires may create a mild degree of inconvenience 
for the subjects. 
 
Length of testing for cognitive circuitry: 
The total length of this task takes a total of 45-60 minutes at baseline and 45 
minutes from week 1 to week 5. There is no expected risk to the subjects beyond 
the ~45-60 minutes of sitting in front of computer for testing and possible 
experience of startle. Every effort will be made to ensure that the subjects are 
comfortable and safe during this period, and the study can be stopped at any time. 
Beyond these issues, there is minimal or no risk to the subjects. 
4.20.2 Benefits 
Study subjects will receive systematic MINI assessment of their DSM comorbidity 
and subthreshold symptoms. This information will be readily available to their 
treating clinicians of PCP if the subjects so desire and agree to disclosure. This 
information might guide long term treatment if subjects are indeed symptomatic. In 
the short-term, the subject will receive close and systematic monitoring for anxiety 
and depressive symptoms and formalized cognitive assessments, beyond current 
standards of care. Easy access to routine physical exams are also a potential benefit.  
 
 
 

 
  
 
                          Page 30 of 39                                                    C O N F I D E N T I A L                                           
.    
 
5   Data Analysis Methods  
5.1  
General Considerations 
The purpose of this study is to assess the change in cerebral blood flow induced by 
the Transcranial Continuous and Pulse Near-Infrared Light Therapy in healthy 
subjects 
 
This study is expected to enroll 30 subjects and treat 10 subjects with TLT for a total 
study duration of 5 weeks. 
5.2  
Specific Primary Aim 
To assess the change in cerebral blood flow induced by the Transcranial Continuous 
and Pulse Near-Infrared Light Therapy in healthy subjects 
 
Hypothesis 1: We anticipate that p-TLT will trigger a significantly greater increase in 
CBF as compared to c-TLT, and c-TLT will trigger a significantly greater effect than 
sham (s-TLT). We expect that we will be also able to estimate the effect size of each 
type of TLT in terms of increase of CBF.  
5.3  
Sample Size 
With respect to the TLT study, we expect 30 subjects to sign informed consent, 
including screen failures.  
 
5.4  
Subject Demographic and Baseline Characteristics 
Subject demographic and baseline characteristics will be summarized.  Descriptive 
summaries will include the number of subjects, mean, standard deviation, median, 
minimum and maximum for continuous parameters, frequencies and percentages for 
categorical parameters. 
5.5  
Efficacy Analyses for Primary Aims 
 
For the primary analysis, delta CBF (measured with DCS-spectroscopy pre- and post- 
TLT) will be compared in all ten subjects by paired t-test. The primary comparisons 
will include: a) delta CBF for p-TLT and delta CBF for c-TLT subjects (n=20 within 
subject comparison) and b) delta CBF for c-TLT and delta CBF for s-TLT (n=20 
within subject comparison).  
5.6 
Safety Analyses 
The number and percent of Adverse Events through study completion will be presented 
by adverse event type. 

 
  
 
                          Page 31 of 39                                                    C O N F I D E N T I A L                                           
.    
 
5.7 
Additional Safety Analyses 
Descriptive statistics for safety endpoints will include number of subjects, mean, 
standard deviation, median minimum and maximum for continuous variables, and 
frequencies and percentages for categorical variables. 
 

 
  
 
                          Page 32 of 39                                                    C O N F I D E N T I A L                                           
.    
 
6 
Ethical Review & Regulatory Considerations 
6.1 
 
Ethical Review 
Prior to the start of the study the Investigator will obtain IRB approval of the protocol 
and the Informed Consent Form. Additional documentation may be required pending 
applicable local requirements.  At least the following documentation must be 
obtained: 
 
1.The IRB/EC approval of the protocol. 
2.The IRB/EC approval of the Informed Consent Form. 
3.The IRB/EC annual (or any other frequency when applicable – i.e. quarterly, 
semiannually according to the local IRB/EC standard operating procedure) 
renewed approval of the protocol. 
4.The IRB/EC approval of any revisions to the Informed Consent Form or 
amendments to the protocol. 
6.2  
Regulatory Considerations  
This study will be conducted in accordance with the Good Clinical Practice (GCP) 
guidelines and other applicable regulatory requirements including but not limited to:  
 
The Food and Drug Administration (FDA) Regulations on Investigational Device 
Exemption (21 CFR 812), 
 
The FDA Regulations on research with human beings (21 CFR 50, 54 and 56),   
 
The Health and Human Services (DHHS) Regulations on research with human 
beings (45 CFR 46 Subparts A, B, C, and D) and 
 
The International Conference on Harmonization (ICH) “Guidance for Industry-E6 
Good Clinical Practice: Consolidated Guideline.”  
The Study will be conducted in the US in accordance with the Privacy Rule (45 CFR 
Parts 160 and 164) of the Health Insurance Portability and Accountability Act of 
1996 (HIPAA).     
 
6.3  
Monitoring Procedures 
Monitoring will be conducted by the PI who will periodically review lab results and 
clinical information. The PI will be coordinating the different phases of the protocol 
implementation such as TLT delivery and follow up. The PI will be frequently in 
contact with the other staff members responsible for TLT delivery and with Litecure 
Inc, which will lend the TLT device. We have minimized risks potentially associated 
with the use of Transcranial Light Therapy (TLT) by requiring two methods of 
contraception (although TLT has not been associated with teratogenic effect).  In 
addition, we are providing the subjects with safety goggles or eye pads during the 
TLT delivery.  We have also included treatment discontinuation rules based on the 

 
  
 
                          Page 33 of 39                                                    C O N F I D E N T I A L                                           
.    
 
development of skin erythema, pain or discomfort lasting more than 24 hours at the 
sites of TLT delivery (to prevent risk of subject’s having an unknown skin 
photosensitivity).    
 
6.3.1 
Training of study site personnel  
Training will be initiated prior to the protocol being implemented. 
Training will consist of both lecture and practicum.  Application of TLT 
procedure will be performed only by PI or his/her designee trained by the 
Sponsor (or its designee) to perform the procedure.  
6.4  
Informed Consent 
The Principal Investigator will be responsible for developing the Informed Consent 
Form The Informed Consent Form will be prepared in accordance with FDA 21 CFR 
Part 50 for all US sites.  The Informed Consent Form will be used to explain in 
simple terms, before the subject is entered into the study, the possible risks and 
benefits to the subject.  The Informed Consent Form will contain a statement that the 
consent is freely given, that the subject is aware of the risks and benefits of entering 
the study, and that the subject is free to withdraw from the study at any time.   
 
Prior to a subject’s participation in the study, the written Informed Consent Form will 
be signed and personally dated by the subject or by an individual authorized to sign 
on behalf of the subject per compliance with local regulations. 
 
If a subject is unable to read or if a legal representative is unable to read, an impartial 
witness will be present during the entire Informed Consent Form discussion.  After 
the written Informed Consent Form and any other written information to be provided 
to the subject is read and explained to the subject or legal representative, and oral 
consent to the subject’s participation in the study has been given by the subject, the 
witness will sign and date the Consent Form.  
 
A witness in the consent process is considered to be an individual that is impartial and 
independent of the study. 
 
Note: 
The Investigator must document acquisition of the written Informed 
Consent Form in the subject’s medical records, and the subject or legal 
representative must be given a copy of the Informed Consent Form document prior 
to enrollment into the study. 
6.5  
Protocol Adherence 
The protocol must be read and followed by all participating study personnel.   
6.6  
Data Collection 
Data will be captured on Case Report Forms - RedCAP. For each subject, a study 
binder will be kept locked in the MGH site. 

 
  
 
                          Page 34 of 39                                                    C O N F I D E N T I A L                                           
.    
 
6.7  
Record Retention 
The Investigator must maintain a file of all documents and records relating to the 
conduct of the study.  Subject files and other source data must be kept for the 
maximum period of time permitted by the hospital, institution or private practice, or 
as specified below.   
 
Study records are subject to inspection by the FDA and other regulatory agencies.   
6.8  
ClinicalTrials.gov 
NIR-FLOW study will be registered with and posted on ClinicalTrials.gov 
(www.clinicaltrials.gov). 
 
 

 
  
 
                          Page 35 of 39                                                    C O N F I D E N T I A L                                           
.    
 
References 
 
1. Mohammed, H. S. Transcranial low-level infrared laser irradiation ameliorates 
depression induced by reserpine in rats. Lasers Med. Sci. (2016). doi:10.1007/s10103-
016-2033-5 
2. Schiffer, F. et al. Psychological benefits 2 and 4 weeks after a single treatment with 
near infrared light to the forehead: a pilot study of 10 patients with major depression 
and anxiety. Behav. Brain Funct. BBF 5, 46 (2009). 
3. Cassano, P., Petrie, S. R., Hamblin, M. R., Henderson, T. A. & Iosifescu, D. V. Review 
of transcranial photobiomodulation for major depressive disorder: targeting brain 
metabolism, inflammation, oxidative stress, and neurogenesis. Neurophotonics 3, 
31404 (2016). 
4. Cassano, P. et al. Near-Infrared Transcranial Radiation for Major Depressive Disorder: 
Proof of Concept Study. Psychiatry J. 2015, 352979 (2015). 
5. Mirzaii-Dizgah, I. et al. Attenuation of morphine withdrawal signs by low level laser 
therapy in rats. Behav. Brain Res. 196, 268–270 (2009). 
6. Hagiwara, S., Iwasaka, H., Hasegawa, A. & Noguchi, T. Pre-Irradiation of blood by 
gallium aluminum arsenide (830 nm) low-level laser enhances peripheral endogenous 
opioid analgesia in rats. Anesth. Analg. 107, 1058–1063 (2008). 
7. Chung, H. et al. The nuts and bolts of low-level laser (light) therapy. Ann. Biomed. 
Eng. 40, 516–533 (2012). 
8. Jagdeo, J. R., Adams, L. E., Brody, N. I. & Siegel, D. M. Transcranial red and near 
infrared light transmission in a cadaveric model. PloS One 7, e47460 (2012). 

 
  
 
                          Page 36 of 39                                                    C O N F I D E N T I A L                                           
.    
 
9. Tedford, C. E., DeLapp, S., Jacques, S. & Anders, J. Quantitative analysis of 
transcranial and intraparenchymal light penetration in human cadaver brain tissue. 
Lasers Surg. Med. 47, 312–322 (2015). 
10. Henderson, T. A. & Morries, L. D. Near-infrared photonic energy penetration: can 
infrared phototherapy effectively reach the human brain? Neuropsychiatr. Dis. Treat. 
11, 2191–2208 (2015). 
11. Konstantinović, L. M. et al. Transcranial application of near-infrared low-level laser 
can modulate cortical excitability. Lasers Surg. Med. 45, 648–653 (2013). 
12. Tian, F., Hase, S. N., Gonzalez-Lima, F. & Liu, H. Transcranial laser stimulation 
improves human cerebral oxygenation. Lasers Surg. Med. 48, 343–349 (2016). 
13. Barrett, D. W. & Gonzalez-Lima, F. Transcranial infrared laser stimulation produces 
beneficial cognitive and emotional effects in humans. Neuroscience 230, 13–23 (2013). 
14. Blanco, N. J., Maddox, W. T. & Gonzalez-Lima, F. Improving executive function 
using transcranial infrared laser stimulation. J. Neuropsychol. (2015). 
doi:10.1111/jnp.12074 
15. Hwang, J., Castelli, D. M. & Gonzalez-Lima, F. Cognitive enhancement by transcranial 
laser stimulation and acute aerobic exercise. Lasers Med. Sci. 31, 1151–1160 (2016). 
16. Naeser, M. A. et al. Significant improvements in cognitive performance post-
transcranial, red/near-infrared light-emitting diode treatments in chronic, mild 
traumatic brain injury: open-protocol study. J. Neurotrauma 31, 1008–1017 (2014). 
17. Naeser, M. A., Saltmarche, A., Krengel, M. H., Hamblin, M. R. & Knight, J. A. 
Improved cognitive function after transcranial, light-emitting diode treatments in 

 
  
 
                          Page 37 of 39                                                    C O N F I D E N T I A L                                           
.    
 
chronic, traumatic brain injury: two case reports. Photomed. Laser Surg. 29, 351–358 
(2011). 
18. Morries, L. D., Cassano, P. & Henderson, T. A. Treatments for traumatic brain injury 
with emphasis on transcranial near-infrared laser phototherapy. Neuropsychiatr. Dis. 
Treat. 11, 2159–2175 (2015). 
19. Zivin, J. A. et al. NeuroThera® Efficacy and Safety Trial-3 (NEST-3): a double-blind, 
randomized, sham-controlled, parallel group, multicenter, pivotal study to assess the 
safety and efficacy of transcranial laser therapy with the NeuroThera® Laser System 
for the treatment of acute ischemic stroke within 24 h of stroke onset. Int. J. Stroke Off. 
J. Int. Stroke Soc. 9, 950–955 (2014). 
20. Hacke, W. et al. Transcranial laser therapy in acute stroke treatment: results of 
neurothera effectiveness and safety trial 3, a phase III clinical end point device trial. 
Stroke J. Cereb. Circ. 45, 3187–3193 (2014). 
21. Lampl, Y. et al. Infrared laser therapy for ischemic stroke: a new treatment strategy: 
results of the NeuroThera Effectiveness and Safety Trial-1 (NEST-1). Stroke J. Cereb. 
Circ. 38, 1843–1849 (2007). 
22. Oron, U., Ilic, S., De Taboada, L. & Streeter, J. Ga-As (808 nm) laser irradiation 
enhances ATP production in human neuronal cells in culture. Photomed. Laser Surg. 
25, 180–182 (2007). 
23. Henderson, T. A. & Morries, L. D. SPECT Perfusion Imaging Demonstrates 
Improvement of Traumatic Brain Injury With Transcranial Near-infrared Laser 
Phototherapy. Adv. Mind Body Med. 29, 27–33 (2015). 
 
 

 
  
 
                          Page 38 of 39                                                    C O N F I D E N T I A L                                           
.    
 
8 
List of Protocol Attachments 
8.1 
 
Protocol attachment 1 – Time Table of Study Procedures 
 
 
 
 
 
 
 
 
 
 
 
 

 
  
 
                          Page 39 of 39                                                    C O N F I D E N T I A L                                           
.    
 
 
 
8.2 
 
Protocol attachment 2 – Schema 
 
 
Week 
1 
2 
3 
4 
5 
 
 
Before 
TLT 
After 
Before 
TLT 
After 
Before 
TLT 
After 
 
Consent 
X 
 
 
 
 
 
 
TLT-
C 
 
 
 
 
 
 
 
TLT- 
SHAM 
 
 
 
 
 
 
 
TLT-
P 
 
 
SCID 
X 
 
 
 
 
 
 
 
Utox 
X 
X 
 
X 
 
X 
 
 
Concomitant 
Medications 
X 
X 
 
X 
 
X 
 
X 
Adverse Event 
X 
X 
 
X 
 
X 
 
X 
ASQ 
X 
X 
 
X 
 
X 
 
X 
SDQ 
X 
X 
 
X 
 
X 
 
X 
Neuro-QoL 
X 
X 
 
X 
 
X 
 
X 
SFI 
X 
X 
 
X 
 
X 
 
X 
SAFTEE 
X 
X 
 
X 
 
X 
 
X 
PANAS 
X 
X 
X 
X 
X 
X 
X 
X 
TSRQ 
 
 
X 
 
X 
 
X 
 
Blinding Scale 
(PBQ) 
 
 
X 
 
X 
 
X 
 
Cognitive Task 
X 
 
 
X 
 
 
X 
 
 
X 
X 
EEG 
 
X 
 
X 
X 
 
X 
X 
 
X