A Multi-Center Trial of the ProLung Test™ (Transthoracic Bioconductance Measurement) as an Adjunct to CT Chest Scans for the Risk Stratification of Patients With Pulmonary Lesions Suspicious for Lung Cancer
Conditions: Solitary Pulmonary Nodule, Multiple Pulmonary Nodules
Sponsor: Fresh Medical Laboratories

Protocol:  PL-208  
Confidential/Proprietary Property of Fresh Medical Laboratories, Inc.                                     Nov. 18, 2016 
 
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Confidential  11/18/2016 1:15 PM  
© 2016 Fresh Medical Laboratories Rev C 
 
 
CLINICAL PROTOCOL 
TITLE: PL-208: 
A 
Multi-Center 
Trial 
of 
the 
ProLung 
Test™ 
(Transthoracic 
Bioconductance Measurement) as an Adjunct to CT Chest Scans for the Risk 
Stratification of Patients with Pulmonary Lesions Suspicious for Lung Cancer 
 
 
   
PROTOCOL NUMBER: 
PL-208  
STUDY TEST: 
ProLung Test 
 
SPONSOR:  
 
 
   Fresh Medical Laboratories™  
 
757 East South Temple, Suite 150 
 
Salt Lake City, Utah 
 
84102  USA 
 
 
801-204.9631 telephone 
 
801-906.0333 facsimile 
 
PL208@prolunglab.com 
 
 
 
  
 
MEDICAL MONITOR: 
Wayne Adams, MD 
 
 
 
 
 
   801-718.3983 
DATE OF PROTOCOL: 
July 2012 
AMENDMENT 2: 
 
 
  July 2016  
 
 
 
 
CONFIDENTIAL INFORMATION 
The information contained herein is confidential and the proprietary property of 
Fresh Medical Laboratories”).  Any unauthorized use or disclosure of such 
information without the prior written authorization is expressly prohibited and 
violates the provisions of the Non-disclosure Agreement between the Sponsor, 
Fresh Medical Laboratories and you and your institution. 
 
 

Protocol:  PL-208 
 
Confidential/Proprietary Property of Fresh Medical Laboratories                                                       Nov. 18, 2016                                
 
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PROTOCOL SIGNATURE PAGE 
 
TITLE: PL-208: 
A 
Multi-Center 
Trial 
of 
the 
ProLung 
Test™ 
(Transthoracic 
Bioconductance Measurement) as an Adjunct to CT Chest Scans for the Risk 
Stratification of Patients with Pulmonary Lesions Suspicious for Lung Cancer 
 
By signing below, the Investigator agrees to adhere to the protocol as outlined and agrees that 
any changes to the protocol must be approved by Fresh Medical Laboratories prior to 
implementing the change or submission of the change for approval by the Institutional Review 
Board (IRB). 
 
This Study will be conducted in accordance with current appropriate local regulations, HIPAA 
(Health Insurance Portability and Accountability Act of 1996) requirements, FDA’s good clinical 
practice regulations (GCPs) ISO 14155: 2011, the Declaration of Helsinki, and local ethical and 
legal requirements. 
 
This study will be listed for disclosure to the extent required by law. 
 
 
Investigator’s Signature:  ________________________________________________ 
 
Printed Name:        _____________________________________________________ 
 
Name of Institution/Company:_____________________________________________ 
 
Date:  ________________________________________________________________ 
 
Medical Data Safety Monitor Signature:______________________________________ 
 
Date:_________________________________________________________________ 
 
Please send a signed copy of this page to Fresh Medical Laboratories via the facsimile to 
801.204-9633 or email mag@prolungdx.com listed on the title page of this protocol PRIOR to 
submission to your institution’s IRB. 
 
 

Protocol:  PL-208 
 
Confidential/Proprietary Property of Fresh Medical Laboratories                                                       Nov. 18, 2016                                
 
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Glossary 
 
AE 
      Adverse Event 
BIA 
      Bioelectrical Impedance Analysis 
 
BP 
      Blood Pressure 
 
PROLUNG     Transthoracic Bioconductance Test 
CRF 
   Case Report Form 
CT 
 
Computed Tomography 
DCF 
   Data Clarification Form 
EKG 
 
Electrocardiogram 
FDA 
   Food and Drug Administration (United States) 
PL 
 
ProLung or Fresh Medical Laboratories Inc.,  or FML 
GCP 
   Good Clinical Practice 
HIPAA 
   Health Insurance Portability and Accounting Act 
ICF 
   Informed Consent Form 
ICH 
   International Conference on Harmonization 
IEC 
   Independent Ethics Committee 
IPL 
Indeterminate Pulmonary Lesions 
IRB 
 
Institutional Review Board 
NCI 
 
National Cancer Institute 
NCN 
Non-calcified nodule 
NSCLC 
 
Non-Small Cell Lung Cancer 
ODA 
 
Optimal Data Analysis 
Operator    
Bioconductance measurement operator 
PENS 
 
Percutaneous Electrical Nerve Stimulation 
PET 
 
Positron Emission Tomography 
PI 
 
Principal Investigator 
RI 
 
Reliability Index 
SAE 
   Serious Adverse Event 
SCLC 
 
Small Cell Lung Cancer 
SDV 
    Source Document Verification 
TENS 
 
Transcutaneous Electrical Nerve Stimulation 
UADE 
 
Unanticipated Adverse Device Effect 
 
 
 

Protocol:  PL-208 
 
Confidential/Proprietary Property of Fresh Medical Laboratories                                                       Nov. 18, 2016                                
 
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TABLE OF CONTENTS 
 
Glossary ............................................................................... 3 
1.0 
Introduction .......................................................................... 6 
1.1         Needed: Immediate and Accurate Risk Stratification for Lung Cancer ....6 
1.2         Fresh Medical Laboratories ProLung Test™ ...........................................6 
2.0 
Study Objectives .................................................................. 7 
2.1 
Objectives and Aims ................................................................................7 
3.0     Study Design .......................................................................... 7 
3.1 
Stabilization Phase ..................................................................................7 
3.2         Validation Phase .....................................................................................8 
3.3 
Safety and Tolerability .............................................................................8 
3.4 
Study Sample Size ..................................................................................8 
3.5 
Assignment to Study Phase ....................................................................9 
3.6         Monitoring................................................................................................9 
3.7         Study Design Summary ...........................................................................9 
4.0 
  Subject Selection and Participation ................................. 9 
4.1  
Inclusion Criteria ......................................................................................9 
4.2  
Exclusion Criteria .................................................................................. 10 
4.3  
Subject Discontinuation ......................................................................... 10 
5.0 
Study Procedures .............................................................. 11 
5.1 
Study Procedures Summary ................................................................. 11 
5.2 
Informed Consent .................................................................................. 11 
5.3 
Inclusion/Exclusion Criteria ................................................................... 11 
5.4 
Demographics and Relevant Health History .......................................... 11 
5.5 
Physical Exam ....................................................................................... 12 
5.6 
Concomitant Medication Assessment ................................................... 12 
5.7 
ProLung Testing Session ...................................................................... 12 
5.8 
Additional Data Collection ..................................................................... 12 
5.9 
End of Study .......................................................................................... 13 
5.10 
Follow-up Outcome Assessment ........................................................... 13 
5.11 
Adverse Event Assessment .................................................................. 13 
6.0 
Adverse Events .................................................................. 14 

Protocol:  PL-208 
 
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6.1 
Adverse Event ....................................................................................... 14 
6.1.1 
Adverse Event Definition ....................................................................... 14 
6.1.2 
Procedures for Adverse Event Reporting .............................................. 14 
6.2 
Serious Adverse Event .......................................................................... 15 
6.2.1 
Serious Adverse Event Definition .......................................................... 15 
6.2.2 
Procedures for Serious Adverse Event Reporting ................................. 15 
7.0 
Data Management .............................................................. 16 
7.1 
Data Collection ...................................................................................... 16 
7.2 
Record Retention .................................................................................. 16 
8.0  
Study Analyses .................................................................. 17 
8.1 
Statistical Analysis Plan ........................................................................ 17 
8.2 
Safety/Tolerability Analyses .................................................................. 18 
8.3   
Baseline and Demographic Data ........................................................... 18 
9.0 
Administrative and Ethical Requirements ........................ 18 
9.1 
Declaration of Helsinki and Ethical Review ........................................... 18 
9.2 
Monitoring Procedures and Auditing of Data ......................................... 18 
9.3 
Subject Information and Consent .......................................................... 19 
9.4 
Subject Data Protection ......................................................................... 19 
9.5 
Financial Disclosure .............................................................................. 19 
9.6 
Changes to the Protocol ........................................................................ 20 
Appendix I: Background Information ........................................... 21 
1.0 
Scientific Literature ................................................................................ 21 
2.0         Clinical Studies: ProLung Test .............................................................. 22 
3.0 
ProLung Unit: Technical Description ..................................................... 25 
3.1         Classifier Algorithm Design and Function .............................................. 25 
3.2         Classification ......................................................................................... 26 
3.3 
Clinical Use ........................................................................................... 26 
3.4 
ProLung Unit Physical Description ........................................................ 26 
3.5 
Safety Information ................................................................................. 26 
Appendix II: References ................................................................ 28 
 

Protocol:  PL-208 
 
Confidential/Proprietary Property of Fresh Medical Laboratories                                                       Nov. 18, 2016                                
 
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1.0 
Introduction 
1.1   Needed: Immediate and Accurate Risk Stratification for Lung Cancer  
Lung cancer is the deadliest form of cancer, with a 5-year survival rate in the United States of 16%i. 
Currently, it is difficult to detect and diagnose lung cancer early enough for treatment to be effective.  
New, immediate, and non-invasive methods are needed to stratify the risk of this disease in its earliest 
stages.  Fresh Medical Laboratories’ bioconductance technology has demonstrated the potential to meet 
these requirements. 
The current standard of care for the detection of lung cancer involves the use of CT chest scans to 
evaluate patients with lesions suspicious for lung cancer.  While these scans provide important 
information about the location and size of lung lesions and raise suspicion of lung cancer, they have 
limited capacity to determine immediately the malignant or benign nature of the lesions they identify.  In 
the screening setting, nearly 25% of all CT screens are “positive” for an indeterminate nodule; among 
these 94% of non-calcified nodules are determined to be benign.ii  This presents a significant diagnostic 
challenge.iii,iv Given the frequency of benign disease, routine biopsy in an indeterminate lesion cannot 
usually be clinically justified.  Because of these constraints, current guidelinesv call for repeated CT scans 
over a period of months or years, in order to detect malignant growth in the detected lesion(s).   
The NCI recently concluded a seminal, multi-center clinical trial in which three repeated CT screenings 
over a two-year period resulted in a 20.0% decrease in mortality for the high risk study cohort.vi,vii  While 
these results are promising, concerns remain about repeated radiation exposure and the persistent high 
rate of false positives, potentially leading to extended evaluation and/or unnecessary biopsy procedures.  
Most importantly, by the time the lesion has grown enough to suggest malignancy, it is too late to achieve 
optimal treatment outcomes.  In addition, there are concerns about the shortage of clinical infrastructure 
to manage the numbers of patients that would screen positive for cancerviii.  The 2011 International 
Association for the Study of Lung Cancer Core Task Force and Screening Implementation Groupix 
recommended the development of new technologies to stratify the risk of cancer patients with 
indeterminate pulmonary nodules.  A fast, accurate and non-invasive technology that could immediately 
stratify the risk of lung cancer in patients with an indeterminate pulmonary lesion could provide invaluable 
life-saving clinical information as well as reduce prolonged anxiety and unnecessary procedures in 
patients with indeterminate lesions.  
1.2   Fresh Medical Laboratories ProLung Test™  
Fresh Medical Laboratories transthoracic bioconductance technology (ProLung Test™) has demonstrated 
the ability to differentiate patients with indeterminate lung lesions as seen by CTx.  The value of this 
innovation lies in its potential to efficiently stratify the risk of lung cancer in a non-invasive, non-radiating 
test that provides immediate results. The Fresh Medical Laboratories ProLung Test has been determined 
to be non-significant risk by previous IRBs.  To date, 160 patients have been evaluated for safety; all 
completed the entire measurement session and none discontinued measurement due to discomfort or an 
adverse eventxi.   
 

Protocol:  PL-208 
 
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2.0 
Study Objectives  
2.1 
Objectives and Aims 
The primary Study hypothesis is that the ProLung Test will demonstrate safety and efficacy in the risk 
stratification of patients with pulmonary lesions identified by CT that are suspicious for lung cancer.   A 
statistically significant result will indicate that patients with a high ProLung Test result have a greater risk 
of developing lung cancer than patients with a low test result.  
There are three Specific Aims of this study: 
1. Optimize and confirm the stability of the ProLung Test risk-stratification algorithm in patients with 
a diagnosis. 
2. Externally validate the efficacy of the ProLung Test risk-stratification algorithm by comparing the 
test result to the conclusive patient diagnosis. 
3. Assess the safety and tolerability of the ProLung Test procedures.  
3.0     Study Design 
This Study consists of two distinct phases, Stabilization and Validation.  The Study will collect data from 
multiple sites (3 to 20), and each site may enroll patients and collect data for the Stabilization and 
Validation Phases with a minimum of three sites for the Validation Phase.  In this multi-center Study, the 
patient’s known diagnosis (of either a malignant or benign outcome) will be compared to the ProLung 
Test results.  The patient’s diagnosis is determined by tissue biopsy, surgical resection or by CT follow-up 
detecting significant nodule growth during a 1 year time frame.  This Study follows the National 
Comprehensive Cancer Network definition for nodule growth.  Nodule growth is defined as: 1) for nodules 
15 mm or smaller: an increase in mean diameter of 2 mm or more in any nodule or in the solid portion of a 
part-solid nodule when compared with the baseline scan, or 2) for nodules 15 mm or more: an increase of 
15% in mean diameter when compared with the baseline scan. Mean diameter is the mean of the longest 
diameter of the nodule and its perpendicular diameterxii.  If no significant growth is detected and the 
nodule is not definitively diagnosed by biopsy or surgical excision, the outcome will be considered benign 
unless indicated otherwise in the patient’s medical record. 
3.1 
Stabilization Phase 
The objective of the Stabilization Phase is to optimize and confirm the stability of the ProLung Test risk-
stratification algorithm in patients with a diagnosis (Specific Aim 1).   In this open multi-center Study, the 
patient’s known diagnosis (of either a malignant or benign outcome) will be compared with the Algorithm.   
Certain components of the Algorithm may be adjusted or optimized to improve overall performance of the 
ProLung Test in the Study population.   
Initially, 100 patients will be enrolled into the initial cohort of the Stabilization Phase.  If required by the 
Sponsor, up to an additional 100 patients, in groups of 25 patients, may be enrolled in the Stabilization 
Phase until the Algorithm achieves stability as determined by the Sponsor.  Once Algorithm stability has 
been determined and confirmed by the Sponsor, the risk stratification Algorithm will be locked for the 

Protocol:  PL-208 
 
Confidential/Proprietary Property of Fresh Medical Laboratories                                                       Nov. 18, 2016                                
 
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Validation Phase.  Data from the Stabilization Phase will also be utilized to assess the safety and 
tolerability of the ProLung Test procedures.  
3.2   Validation Phase 
The objective of the Validation Phase is to externally validate the efficacy of the ProLung Test risk-
stratification Algorithm by comparing the test result to the conclusive patient diagnosis (Specific Aim 2).  
The risk stratification Algorithm result is recorded and locked for this Phase. 
The Validation Phase is blinded as follows:  
Sponsor’s Statistician is blinded from the patient’s diagnosis outcome (malignant, benign or 
inconclusive).  The patient’s ProLung Test results will be provided to the Sponsor and/or 
Statistician. 
Site clinical study team, including the PI, study coordinators, and ProLung Test operators, 
is blinded from the ProLung Test risk stratification algorithm results.  At each site, the patients’ 
diagnoses are accessible by the site clinical study team.   
Initial Study enrollment in the Validation Phase will be 100 patients.  To maintain the Study’s statistical 
power, enrollment will continue until there is both a minimum of 100 subjects and a minimum of 40 
ProLung high risk and a minimum of 40 ProLung low risk subjects are enrolled.   If required by the 
Sponsor and/or Statistician, up to an additional 50 patients, in groups of 25 patients, may be enrolled in 
the Validation Phase prior to breaking the statistical blind.  The statistical blind is broken at the end of 
Study recruitment and monitoring with approval by the Sponsor and/or Statistician. 
If required by the Sponsor and/or Statistician, up to an additional 35 subjects may be enrolled in either the 
Stabilization or Validation Phase to replace the enrolled subjects that were lost to follow-up or fail the 
Study Inclusion / Exclusion Criteria.   
3.3 
Safety and Tolerability 
To assess the safety and tolerability of the ProLung Test procedures, the Study will utilize the data from 
all patients enrolled in the Stabilization and Validation Phases.  Information collected about adverse 
events and tolerability will be analyzed as described in sections 5.8 and 6.0 (Additional Data Collection 
and Adverse Events).  
3.4 
Study Sample Size 
This Study may enroll a maximum of 200 in the Stabilization Phase and 150 in the Validation Phase.  The 
Study may enroll a minimum of 100 in the Stabilization Phase and 100 in the Validation Phase.  In the 
Validation Phase, the sample size has been determined to provide greater than 90% power to reject the 
primary study hypothesis. 

Protocol:  PL-208 
 
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3.5 
Assignment to Study Phase 
The site study teams will enroll patients sequentially into the Stabilization Phase.  When the Sponsor has 
confirmed the stability of the ProLung Test risk-stratification Algorithm, the Stability Phase will be 
completed and the Validation Phase launch will be confirmed by written notice to each site clinical study 
team.   Prior to enrollment in the Validation Phase, Study blinding will be established and monitored. 
3.6     Monitoring  
 
The study Sponsor will provide monitors to monitor Protocol compliance and provide guidance pertaining 
to the management of the Study.  
3.7    Study Design Summary  
 
Design Element 
Stabilization Phase 
Validation Phase 
Patient Enrollment  
100-200 patients   
100-150 patients  
Sites 
3-20 
3-20 
Blinding and Breaking the 
Statistical Blind 
Open phase - Optimize 
algorithm with known 
diagnosis 
Blinded phase – Sponsor’s 
Statistician is blinded to 
diagnosis and site study team 
is blinded to ProLung Test 
results. The statistical blind is 
broken at the end of Study 
recruitment and monitoring 
with approval by the Sponsor 
and/or Statistician. 
Primary Objectives: 
Stability of ProLung Test risk 
stratification algorithm as 
confirmed by Sponsor. 
Clinical efficacy in the risk 
stratification of patients with 
indeterminate lesions. 
Safety and tolerability 
Safety and tolerability 
 
4.0 
  Subject Selection and Participation 
4.1  
Inclusion Criteria  
Subjects who meet all of the following criteria may be enrolled in this Study: 
1. Subject is male or female, age 18 or older. 
2. Subject has undergone CT scan of the lung(s) that indicates one or more nodules or lesions 
suspicious for lung cancer. 
3. Subject’s pulmonary nodule or lesion is greater than 4mm. Size is determined by the largest 
nodule or lesion dimension identified from CT imaging. 
4. Subject meets one or more of the following conditions: 
o 
indicated for a tissue biopsy  

Protocol:  PL-208 
 
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o 
indicated for surgical resection of the lung 
5. Subject must be able to receive a ProLung Test 
o 
within 60 days of abnormal CT (Inclusion Criterion 2 & 3)  
o 
within 60 days prior to the tissue biopsy or surgical resection (Inclusion Criterion 4). 
6. Subject is capable of understanding and agreeing to fulfill the requirements of this Protocol. 
7. Subject has signed the IRB/IEC approved Informed Consent Form (“ICF”). 
4.2  
Exclusion Criteria  
The following criteria will disqualify a subject from enrollment into this Study: 
1. 
Subject has an implanted electronic device in the chest.  
2. 
Subject receiving therapy for suspected chest infection such as fungal infection or 
tuberculosis. 
3. 
Subject with diagnosed malignancy other than lung cancer, non-melanoma skin cancer or 
any cancer in which the Principal Investigator does not suspect metastatic disease to the 
lung, who has 2 or more suspicious pulmonary nodules.  
4. 
Subject has received an invasive medical or surgical procedure within the thoracic cavity 
within 30 days prior to the ProLung Test or within the previous 14 days for a bronchoscopic 
procedure. 
5. 
Subject presents with an anomalous physical or anatomical condition that precludes ProLung 
Test measurement. 
6. 
Subject will have undergone unusually strenuous exercise within 24 hours.  
7. 
Subject who has significant systemic diseases such as uncontrolled diabetes, advanced 
heart failure, or a recent myocardial infarction, or other medical condition such as severe 
morbid obesity that in the judgment of the Principal Investigator would make him/her 
unsuitable for the Study. 
4.3  
Subject Discontinuation  
In accordance with the Declaration of Helsinki, subjects are free to discontinue the Study at any time for 
any or for no reason.  Should a subject decide to withdraw, every effort should be made to obtain 
information about the reason(s) for discontinuation.  The Principal Investigator also has the right to 
withdraw subjects from the Study at any time at his/her discretion.   
4.3.1 Events Necessitating Premature Discontinuation 
The occurrence of any one of the following events may necessitate premature discontinuation of a subject 
from the Study: 
• 
Subject develops any of the exclusion criteria after enrollment. 
• 
Subject is unwilling to continue in the Study. 
• 
Subject does not comply with the Study Protocol. 
• 
Subject being followed by CT for nodule growth who does not receive a follow-up CT by their 1-year 
enrollment anniversary or within 60 days after their 1-year enrollment anniversary. 

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• 
Principal Investigator decision. 
4.3.2 Events Eliminating Subjects from Primary Analyses 
• 
Incomplete or lost medical records. 
• 
Inconclusive biopsy result. 
• 
Incomplete ProLung Test data. 
5.0 
Study Procedures 
5.1 
Study Procedures Summary 
 
Patient Visit:  
• 
Obtain demographics 
• 
Obtain smoking history, and details of industrial exposure  
• 
Obtain recent medical and surgical history 
• 
Record concomitant medications (most recent 2 weeks)  
• 
Conduct physical examination 
• 
Perform ProLung Test.  Adverse Event assessment 
• 
Administer Tolerability Questionnaire  
• 
Print out ProLung Test summary report for inclusion in patient’s source documentation 
Patient Data and Source Documentation Collection: 
• 
Transmit patient data to Sponsor weekly as patients complete ProLung Testing 
• 
Obtain radiological reports including CT, chest x-ray, follow-up CT, PET, and MRI.  
• 
Obtain surgical report 
• 
Obtain biopsy report 
• 
Obtain pathology report 
5.2 
Informed Consent 
The Informed Consent must be documented on the current IRB-approved version of the informed consent 
form.  The consent process must also be documented in the site’s Study Notebook.  The Informed 
Consent must be obtained prior to performing any Study-specific procedures.  The patient must be given 
a fully executed copy of the informed consent form. The site retains the original signed informed consent 
form. 
5.3 
Inclusion/Exclusion Criteria 
All inclusion and exclusion criteria must be evaluated prior to subject enrollment and at the start of the 
ProLung Test protocol.  Documentation that the subject meets all of the inclusion criteria and none of the 
exclusion criteria must be present in the subject’s source documents in Site Study Notebook.  If an 
exemption to the inclusion or exclusion criteria is granted by the Sponsor, such exemption will be 
recorded in the CRF.  
5.4 
Demographics and Relevant Health History 
 

Protocol:  PL-208 
 
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a. The subject’s gender, age, and ethnicity (patient’s optional response) will be recorded.   
b. The subjects will be asked about their smoking history including pack-years smoking and how 
recently they discontinued smoking if applicable. 
c. The subjects will be asked about their past and current exposure to cancer-causing agents. 
d. The subjects will be asked about their current pulmonary symptoms. 
e. A brief medical and surgical history will be obtained.  The medical history should include all 
significant current conditions, including recent illnesses and history, within two years of the time of 
the ProLung Test.  The surgical history will include any major surgeries performed within five 
years of the date the ProLung Test is performed. 
5.5 
Physical Exam 
A physical examination will be performed at the patient visit to assess the patient’s general physical 
status. Physical characteristics of height, weight will be measured and recorded.   
5.6 
Concomitant Medication Assessment 
The subject’s usage of concomitant medications within the most recent 2 weeks will be recorded including 
name of the drug, dosage, and reason for use.  Medications will include both prescription and over-the 
counter drugs, but will EXCLUDE herbal or nutritional supplements.    
5.7 
ProLung Testing Session 
The ProLung Test operator non-invasively measures certain anatomical locations on the surface of the 
skin.  ProLung Test locations are displayed immediately prior to measurement and sequenced for the 
Operator performing the test.  Real-time assessment of the quality of each measurement assists the 
operator to capture measurement data. 
All operators involved in measuring subjects will have previously been trained and certified in use of the 
ProLung unit as provided by the Sponsor and described in detail in the Operator Training Manual. They 
will demonstrate competence based on criteria established by the Sponsor prior to using the ProLung 
Test on enrolled patients.  The ProLung Test requires an average of 20 minutes.  In previous clinical 
studies, time required to complete the ProLung Test ranged from 16 to 23 minutes. 
Patient ProLung measurement data will be printed out at the end of each test session and placed into the 
subject’s source document file. This print-out only contains summary information of the collected data and 
does not represent a complete data-file that is analyzed with the Algorithm.  Weekly, ProLung Test data 
will also be stored to a Sponsor provided USB flash drive which is used to send ProLung Test data to the 
Sponsor through a secure Study file sharing website.    
5.8 
Additional Data Collection  
 
Post-Measurement (following ProLung Test) 
• 
Adverse Event assessment 

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• 
Tolerability Questionnaire: Following the ProLung Test, subjects will be asked to complete a brief 
questionnaire regarding their experience and perception of the procedure. The questions are:  
1. Did the test cause any discomfort? If so, please describe.  
2. Did the time required for measurement seem too long? If so, what amount of time 
seems reasonable?  
3. Would you agree to undergo measurement again? If not, what is the reason?  
4. Do you have any suggestions for improving the measurement procedure? If so, please 
describe.  
5.9 
End of Study 
The subject’s active participation in the Study is completed once the ProLung Test and the adverse event 
assessment have been completed.  The subject’s active participation concludes prior to the performance 
of the lung biopsy.  
5.10 Follow-up Outcome Assessment 
 
The outcomes of any ancillary diagnostic procedures conducted as part of normal clinical care will be 
collected for further analysis and the provided CRF page(s) will be completed as appropriate.   The Study 
participant will sign a “Release of Medical Information” at the time of the consent process. All laboratory 
and radiological tests performed to further evaluate the abnormal chest CT will be obtained by the study 
team and kept as part of the participant’s source document. This will include but are not limited to results 
of biopsies, sputum cytology, repeat chest CT scans, repeat chest radiography, MRI and PET scans.  
 
Prior to the close of the Study, each site will obtain the most recent CT Report and History and Physical (if 
available) for all enrolled subjects who received a benign biopsy result or a CT follow-up showing stability 
to re-confirm that the subjects have remained free of malignancy. 
 
5.11 Adverse Event Assessment 
During, and immediately following the ProLung Test, subjects will be observed for any potential adverse 
event.  Any unanticipated and unfavorable sign, e.g., a symptom described by the patient or noted by the 
site staff will be recorded as an adverse event in the source documents and CRF, and followed-up with 
more specific questions or actions as required.   

Protocol:  PL-208 
 
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6.0 
Adverse Events 
6.1 
Adverse Event  
6.1.1 Adverse Event Definition 
An adverse event is defined as any untoward medical occurrence in a patient or clinical investigation 
subject involved in the testing of a medical device and which does not necessarily have to have a causal 
relationship with the device. 
An adverse event can therefore be any unanticipated and unfavorable sign, symptom or disease 
temporally associated with the use of a device, whether or not considered related to the product. 
An AE includes: 
• 
An exacerbation of a pre-existing illness or symptom 
 
• 
Post-measurement (i.e. after the ProLung Test) events that occur as a result of protocol-
mandated procedures  
• 
A condition detected or diagnosed after device utilization 
An AE does not include: 
• 
The disease or disorder being studied or signs and symptoms associated with the 
disease/disorder unless there is an unexpected worsening of the subject’s condition 
• 
A pre-existing disease or condition present at the start of the Study that did not worsen 
• 
Elective medical or surgical procedures unless resulting from a SAE 
6.1.2 Procedures for Adverse Event Reporting 
Adverse events will be recorded from the time the subject has begun measurement with the device 
through Study completion.  Any change in medical status or untoward event that occurs from the time the 
subject signs informed consent and undergoes ProLung measurement will be recorded as a baseline 
condition in the medical history. 
Any unanticipated and unfavorable sign (e.g., a symptom or physical anomaly described by the subject or 
noted by the site staff) will be recorded as an adverse event.  Changes in the subject’s physical or mental 
condition or other responses must be reported as AEs if they are considered clinically significant by the 
Principal Investigator, if they fulfill SAE criteria or if they cause discontinuation from the Study.  
AEs not resolved at the end of the ProLung Test will be followed after clinical Study completion or as long 
as medically relevant as judged by the Principal Investigator.  
The Sponsor will conduct an evaluation of any unanticipated adverse device effect (UADE) in accordance 
with local regulations and report the results of the evaluation to the Investigator, IRB and any necessary 
authorities within 10 working days after the Sponsor first receives notice of such an event.  

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6.2 
Serious Adverse Event  
6.2.1 Serious Adverse Event Definition 
An adverse event is considered serious if it: 
• 
Occurs during Study measurement (ProLung Test) or immediately thereafter and, 
• 
Meets any of the criteria outlined in the following section 
An SAE is any adverse event that results in any of the following outcomes: 
• 
Death or is life-threatening: A life-threatening SAE is any SAE that places the subject in the 
opinion of the Principal Investigator, at immediate risk of death from the reaction as it occurred.  It 
does not include a reaction that, had it occurred in a more serious form, might have caused death 
• 
Persistent or significant disability/incapacity 
• 
Inpatient hospitalization or prolongation of existing hospitalization 
Important medical events that may not result in death, be life-threatening, or require hospitalization may 
be considered a serious adverse event 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. Scheduled and/or elective hospitalizations will not be defined as serious adverse 
events for this clinical Study. 
6.2.2 Procedures for Serious Adverse Event Reporting 
The Principal Investigator must inform the Sponsor and the Medical Monitor within 24 hours of becoming 
aware of any SAE that occurs during the course of the clinical Study. The Principal Investigator may 
communicate the event to the Sponsor or Medical Monitor; however, a completed SAE form must be 
faxed or scanned and emailed to the Medical Monitor and to the Sponsor within 24 hours of the initial 
reporting. All serious events must be reported, whether or not they are considered causally related to the 
device utilization. Appropriate clinical, diagnostic, and laboratory measures must be performed to 
delineate the cause of the SAE in question and the results reported. All tests that reveal an abnormality 
that is considered device related should be repeated at appropriate intervals until:  
• 
The cause is determined 
• 
A return to baseline value occurs 
• 
Stable results are obtained over 2 to 3 consecutive readings that are clinically acceptable and 
safe for the subject 
The Principal Investigator is required to assess the causal relationship to the Study device for each SAE 
to determine if the event is associated with the use of the device.  Events should be classified as 
associated with the use of the device if there is a reasonable probability that the experience may have 
been caused by the device.  
The Sponsor will conduct an evaluation of any UADEs in accordance with 21 CFR 812.46(b) and report 
the results of the evaluation to the Investigator, IRB and FDA within 10 working days after the Sponsor 
first receives notice of the effect in accordance with 21 CFR 812.150(b(1)). The Sponsor will terminate the 
investigation when a determination has been made that an unanticipated adverse device effect presents 

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an unreasonable risk to subjects. Termination will occur no later than 5 working days after the Sponsor 
makes the determination and not later than 15 working days after the sponsor first received notice of the 
effect in accordance with 21 CFR 812.46(b)(2). 
7.0 
Data Management 
7.1 
Data Collection 
The site study team will make data entry on the case report forms (CRF)s and keep records of the 
subject’s visit in the source documents for that site.  The Principal Investigator or designee will be 
responsible for the timely recording of patient data. 
To ensure that correct data have been entered on the CRF, CRFs will be 100% source-data verified by a 
Monitor from the Sponsor, who will notify the Principal Investigator or designee regarding questions or 
missing data.  When data have been entered in the CRF, signed by the Principal Investigator, and all 
questions and/or corrections have been resolved and verified, the original of each CRF page(s) will be 
collected by the Sponsor.  The PI will retain a copy of completed and monitored CRF pages.  Any 
remaining questions or missing data subsequent to retrieval of a CRF page from a site will be noted on 
Data Clarification Forms (DCFs), which will be sent to the Principal Investigator.  DCFs should be 
completed by the Study site and signed by the Principal Investigator in a timely manner. 
ProLung Test subject data will be sent to the Sponsor on a weekly basis as subjects are enrolled.   
7.2 
Record Retention  
All records relating to the conduct of the Study are to be kept in a locked and secure location and held by 
the Principal Investigator until notified by the Sponsor that the records may be destroyed. Essential 
documents should be retained until at least 2 years after the last approval of a marketing application in an 
ICH region and until there are no pending or contemplated marketing applications in an ICH region or at 
least 2 years have elapsed since the formal discontinuation of clinical development of the investigational 
product.  These documents should be retained for a longer period, however, if required by the applicable 
regulatory requirements or by agreement with the Sponsor.  The Sponsor will notify the Principal 
Investigator of the retention period required to satisfy appropriate regulations. If the Sponsor has not 
notified the Principle Investigator and 10 years has passed since the last subject was enrolled, all records 
may be destroyed. 
The Principal Investigator will allow representatives of the Sponsor’s monitoring team and of the 
appropriate regulatory authority to inspect all Study records, CRFs, and corresponding portions of the 
Study patients’ office and/or hospital medical records at regular intervals throughout the Study and after 
the Study has completed if required.  These inspections are for the purpose of verifying adherence to the 
protocol, the completeness and exactness of the data being entered in the report forms, and compliance 
with regulations. 

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8.0  Study Analyses  
8.1 
Statistical Analysis Plan 
 
Stabilization Phase 
Stabilization Phase is not blinded.  The malignancy or benignity status of the patients will be available as 
described in Section 4.0 Study Design.  Algorithm stabilization will be tested by the Sponsor against prior 
clinical results and across study patients.  In addition, in this phase, the Sponsor and Statistician may 
evaluate the impact of demographic, geographic and anatomical factors on the stability of the risk 
stratification Algorithm.  Certain inter and intra patient bioconductance factors may also be evaluated for 
impact on Algorithm stability.  
The statistical approach to the risk stratification Algorithm stabilization will be to modify the Algorithm 
using Optimal Data Analysis (ODA) programmed in Stata™ version 11 statistical software that allows a 
large number of variables at a time while treating each measurement variable independently.  This will be 
in contrast to traditional multivariable regression that treats variables as possibly correlated, but only 
allows for a small number of predictor variables at a time which will also be used to optimize the 
Algorithm.  Variable selection, cut-point determination, decision nodes, and so on, will be based on 
variables that provide improved discrimination and validate successfully. 
After stability of the risk stratification has been confirmed by the Sponsor, the Algorithm will be locked, 
and further changes will not be permitted.  
 
Validation Phase 
 
The locked risk stratification Algorithm will be applied to the patients participating in the Validation Phase 
in which the Sponsor’s Statistician is blinded to the actual patient diagnosis.   
With respect to determining the optimal sample size of the Validation Phase, it is assumed that the 
sensitivity of the ProLung test (probability of having had a high risk test among subjects who 
subsequently had a malignant biopsy) is 70% and that the specificity (probability of having had a low risk 
test among subjects who subsequently had a benign biopsy) is also 70%.  It is further assumed that 
among the patient population studied in this Study, approximately 70% will have a malignant biopsy.  
Based on these assumptions, 88 subjects will provide 90% power to reject the primary null hypothesis.  
The primary analysis will only include subjects who have a biopsy within 60 days of their ProLung test 
and have a definitive biopsy result.  Subjects who fail to have a biopsy or do not have a definitive biopsy 
result will be excluded.  To account for as many as 12% of such subjects, 100 subjects will be enrolled 
into the Validation Phase.  Furthermore, to maintain the Study’s statistical power, enrollment will continue 
until there is both a minimum of 100 subjects and a minimum of 40 ProLung high risk and a minimum of 
40 ProLung low risk subjects are enrolled. 
The primary null hypothesis is that the relative risk of a malignant biopsy among subjects with a high risk 
ProLung test result to that of subjects with a low risk ProLung test result is 1.0, vs. the alternative 
hypothesis that the relative risk is different from 1.0.  This hypothesis will be tested by a two-sided 
Fisher’s Exact Test at the 5% level of significance.  If the relative risk is found to be greater than 1.0 

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(p<0.05), it will be concluded that subjects with a high risk ProLung test result are prognostically at 
greater risk to have a malignant biopsy than subjects with a low risk ProLung Test result.  The relative risk 
will also be presented with its approximate 95% confidence interval. 
8.2 
Safety/Tolerability Analyses 
Subject data on AEs and SAEs will be collected during and immediately following the ProLung Test 
period.  Treatment emergent AEs will be summarized using descriptive statistics (e.g., number and 
percentage of patients).  Treatment emergent AEs are defined as AEs whose onset occurs, severity 
worsened or intensity increases after undergoing the ProLung Test.  We will survey the subjects for their 
comfort and tolerance of the overall test procedure. 
8.3   Baseline and Demographic Data 
Patient information including age, gender, smoking history, occupational exposure, and ethnic 
background (when available) will be collected and used to summarize the baseline and demographic 
characteristics for each Cohort and overall.  
9.0 
Administrative and Ethical Requirements 
9.1 
Declaration of Helsinki and Ethical Review 
The Study will be performed in accordance with the principles stated in the Declaration of Helsinki, FDA 
GCP regulations and ISO 14155: 2011 requirements.  The final Study protocol, including the final version 
of the Informed Consent Form (ICF) to be used, must be approved by an IRB before enrollment of any 
patients in the Study.  The opinion of the IRB will be dated and given in writing.  A list of those present at 
the committee meeting (names and positions) should be attached. The regulatory authority and/or IRB 
will be informed of any SAEs and amendments to the protocol according to local and FDA requirements.  
All correspondence with the IRB must be filed by the Principal Investigator and a copy forwarded to the 
Sponsor.  
9.2 
Monitoring Procedures and Auditing of Data 
A Study Monitor will review the Study data during, and at the end, of the investigation.  All personnel 
involved in the Study should be prepared to make time to discuss the data with the Monitor. 
The following will be reviewed by the Study Monitor: 
• 
Subject enrollment 
• 
Compliance with the protocol 
• 
Informed consent verification 
• 
Completion of the CRFs and the correction of any missing or incorrect data 
• 
100% Source document verification (SDV) 
• 
Adverse event identification and reporting 

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• 
Contents of the Investigator File and Study Documents 
In accordance with Good Clinical Practices (GCP), the Monitor will undertake auditing of data recorded on 
the CRF against the source documents.  The purpose of source document verification is to verify, so far 
as is possible, that the information on the CRF reflects the data recorded in the patient’s hospital/clinic 
chart.  SDV will be performed with due regard for subject confidentiality and in accordance with local 
privacy regulations, and HIPAA requirements if any.  
9.3 
Subject Information and Consent 
The Principal Investigator or designee will ensure that the subject is given full and adequate verbal and 
written information about the nature, purpose, possible risks and benefits of the Study.  Subjects must 
also be notified that they are free to stop participating in the Study at any time without prejudice to the 
quality or level of care provided.  Subjects will be informed that the effects of the device will be followed 
throughout the Study.  The subject should be given the opportunity to ask questions and time to consider 
their participation. The Principal Investigator or designee is responsible for obtaining and documenting 
written informed consent from all subjects before enrollment.  The subject must be given a copy of the 
fully executed ICF.  The Principal Investigator must retain an original fully executed ICF.    
9.4 
Subject Data Protection 
All data provided to the Sponsor will be identified by a unique subject number, thereby ensuring that the 
subject’s identity remains unknown.  The subjects should be informed in writing, that their data may be 
stored and analyzed in a computer, with confidentiality maintained in accordance with local regulation. 
The subjects should also be informed in writing that authorized representatives of the Sponsor and/or 
regulatory authorities may require access to those parts of the chart records relevant to the Study, 
including medical history, for data verification. 
The Principal Investigator is responsible for keeping a subject identification list of all subjects screened 
and enrolled which includes the following information: subject number, full name, and a secondary unique 
identifier (i.e., hospital/clinic number).  
9.5 
Financial Disclosure  
All investigators participating in clinical studies must comply with local and national financial disclosure 
regulations if applicable.  If required, these are to be submitted in support of an application for market 
approval.   
The financial information typically disclosed includes four types of data.  They are: 
i) significant equity interest in the Sponsor held by the Investigator equal to or exceeding US 
$50,000  
ii) any financial arrangement entered into between the Sponsor and the investigator whereby the 
value of the compensation to the investigator for conducting the Study could be influenced by the 
outcome of the Study 

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iii) compensation, other than that which is associated with conducting the referenced Study, paid 
by the sponsoring company to an investigator, exclusive of the costs of conducting the clinical 
Study or other sponsored clinical studies (e.g., a grant to fund ongoing research, compensation in 
the form of equipment or retainers for consultation or honoraria, etc.) 
iv) proprietary interest in the product tested in the covered Study held by an investigator.  
To comply with this regulation, investigators are required to complete the financial disclosure form which 
will be supplied at site initiation.  These disclosure requirements cover Principal Investigators, co-
investigators and sub-investigators and their spouses and dependent children.  The originals of these 
documents must be included in the Principal Investigator’s regulatory documents. 
9.6 
Changes to the Protocol 
No change in the Study procedures shall be effected without the mutual agreement of the Sponsor and 
the Principal Investigator. All changes must be documented as signed protocol amendments, or as a 
revised protocol.  Changes to the protocol require notification to or approval by the IRB and the regulatory 
authorities before implementation.  Local regulatory requirements must be followed. 
The Sponsor is responsible for the distribution of protocol amendment(s) to the Principal Investigator(s) 
and those concerned within the conduct of the Study.  The Principal Investigator is responsible for the 
distribution of all amendments to the IRB and all staff concerned at his/her center. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Appendix I: Background Information 
1.0 
Scientific Literature 
Bioelectrical Properties of Living Tissue  
By definition, electrical impedance is the ratio of the voltage difference to the current across a circuit or a 
body (Ohm’s law), and conductance is the inverse of impedance (1/impedance).  The dielectric properties 
of human cells and tissue are widely recognized and are essential for several diagnostic 
procedures currently in use, such as the electrocardiogram.xiii Bioelectrical conductance is the basis for 
the monitoring of certain physiological processes such as “impedance plethysmography” for measuring 
blood volume or Electrical Impedance Tomography” (EIT) for assessing lung ventilationxiv xv.  Other 
examples 
include 
multi-frequency 
Bioelectrical 
Impedance 
Analysis 
(BIA) 
to 
measure 
the 
electrophysiological properties of cell membranes and related body tissues as a predictor of body weight, 
fat content, and extracellular and intracellular water in a wide range of medical conditions.xvi  
Bioconductance and Malignant Tissue 
Several experimental studies have demonstrated that the electrical properties of cancerous tumors vary 
significantly from normal, nonmalignant tissue
xviii
xxiii
xvii.  The mechanism has been attributed to the high water 
and sodium content within cancerous tissues with movement of potassium, magnesium and calcium out 
of the cell
,xix. Other possible contributors include not only altered membrane permeability but also 
changes in membrane composition, the nucleus to cytoplasm ratio as well as alterations in cellular 
composition and densityxx xxi.  Many of the published studies are in breast cancer.  For example, one 
study examined a set of 120 impedivity spectra collected in breast tissue immediately after excision from 
64 patients undergoing breast surgeryxxii.  This investigation reported that no significant differences were 
found between groups of normal breast tissue and those consisting of benign pathology such as 
fibroadenomas.  However, the bioelectrical properties of the carcinoma group differed from the normal 
and benign tissue groups
.  In vitro, hepatic studies have found differential electrical conductivity 
changes between tumor and control tissue in a rat modelxxiv.  Another in vitro study examined brain, 
breast, gastric and colon cancers that had been implanted in a nude mouse model, grown, and then 
excised for investigation of electrical conductance propertiesxxv.  This investigation found that all four 
cancerous tissues had conductance values that were clearly different from non-cancerous tissuexxvi.   
In summary, studies conducted at the cellular and tissue level (as well as on recently excised human 
tissue) have demonstrated differences in electrical conductivity associated with various disease states 
including cancer. 
Bioconductance and Cancer Diagnosis 
Many clinical investigations have examined the potential of using electrical properties to aid in cancer 
diagnosis.  Electrical bio-conductance has been shown to successfully discriminate between cancerous 
and non-cancerous tissue for skin cancer,xxvii
xxviii
 cancer of the lymph nodes
 and breast cancer xxix and has 
shown promise as a screening tool for the detection of cervicalxxx and colorectal cancersxxxi. 

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There is also growing evidence regarding differences in lung tissue bioconductivity associated with lung 
cancer.xxxii
xxxiii.  The conclusion drawn by Toso and colleagues is that a distinct 
difference in transcutaneous bioimpedence (a combination of resistance and capacitance) can be seen 
between patients with advanced lung cancer and normal controls, and that in the advanced cancer 
cohort, regardless of their measured Body Mass Index
  In one non-invasive study of bioconductance measurement in lung cancer, bioelectric 
impedance vector analysis in patients with known advanced stage (IIIB and IV) lung cancer (66) was 
compared with healthy controls (56)
 (or BMI, one commonly used index of nutrition), 
bioconductance measurement differences provided a better measurement of survival, and hence 
prognosis.  
A second study utilized electrical impedance tomography to provide in vivo imaging prior to surgery in 22 
patients with single sided lung cancer and 7 healthy subjectsxxxiv.  These investigators found that “images” 
in 19 of the 22 affected lung cancer patients showed differences in conductivity that were statistically 
different from the average conductivity of a healthy lung.   
In addition to these studies, the ProLung Test developed by Fresh Medical Laboratories has been 
examined in 3 clinical studies, which evaluated the feasibility of using a non-invasive bioconductance test 
to discriminate between malignant and non-malignant lung tissue. 
  2.0    Clinical Studies: ProLung Test  
 
The initial feasibility study, FML-201, tested the ProLung Test to discriminate between the bioconductance 
measurements recorded for age-and gender-matched subjects with and without biopsy-confirmed lung 
cancer. The second study, FML-203, examined measurement reliability and reproducibility in a normal 
subject sample.  The third, FML -204, tested the ProLung Test’s discriminatory ability in undiagnosed 
patients who exhibited lesions that were suspicious for lung cancer. 
 
A. Feasibility Study (FML-201). McHenry IL 2007. The study’s first Specific Aim was to investigate 
ProLung Test effectiveness in discriminating between clinically symptomatic patients (n=18) with and 
without (n=18) lung cancer.  The subjects were also evaluated for any adverse events connected with use 
of the test. 
 
The study was a single-site, single-blind (i.e., ProLung Test operator and statistician were unaware of 
subject diagnosis), two-arm design with screening and bioelectrical conductance measurement visits. 
Biopsy was performed prior to conductivity measurement.  Figure 1 below illustrates the performance of 
the optimal algorithm in this initial set of patients. 
 

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Figure 1. Discrimination of cancer from non-cancer patients.  
 
Results.  In the FML-201, the derived Algorithm was able to perfectly discriminate (vertical line) biopsy-
confirmed cancer subjects from age- and gender-matched cancer-free subjects.  X-axis: composite score 
units. Y-axis:  study arm 1, non-cancer; study arm 2, cancer. 
 
Conclusion. These data supported the hypothesis that trans-thoracic conductance measurements 
obtained using the ProLung Test distinguish statistically significant conductivity differences between 
cancerous and non-cancerous lung tissue in a small patient sample.  This suggested feasibility of the 
technology. Further algorithm development with validation was planned for a subsequent larger study. 
 
B. Reliability and Reproduceability Study (FML-203).  The variability of repeated ProLung conductance 
measurements was investigated using a single ProLung unit and operator on 22 normal subjects over two 
days of testing at the Salt Lake Community College in Salt Lake City Utah. The Specific Aim was to 
investigate ProLung unit variability as estimated from repeated measurements on the same subject in 
succession.   
Results.  The maximum and minimum conductance results were comparable, with slightly lower standard 
deviations for maximum conductance readings and extremely high reliability indices for both measures.  
For both data sets, the same measurement points were found to have minimal variability (and maximal 
reliability) indices. 
Conclusion.  The results suggest a high degree of consistency in the ProLung measurements and support 
the reliability of this procedure as an adjunct to other methods used to risk-stratify patients who have 
suspicious lung lesions. 
 
C. Transcutaneous Computed Bioconductance Measurement in Lung Cancer: A Treatment 
Enabling Technology Useful for Adjunctive Risk Stratification in the Evaulation of Suspicous 
Pulmonary Lesions 
 
Methods..  The research was conducted at Johns Hopkins School of Medicine, by members of the faculty 
at that institution and the University of Utah School of Medicine, as well as members of the Fresh Medical 
Laboratories clinical staff.  The Study was published in the April 2012 edition of the Journal of Thoracic 
Oncology. The purpose of this Study was to further develop a mathematical algorithm that discriminates 
between benign and malignant pulmonary lesions found on spiral chest CT scans, using the noninvasive 
transcutaneous computed bioconductance (ProLung) measurement Test. The Study’s hypotheses were 
(1) designed to assess the stability of the ProLung Test classification Algorithm when used as an adjunct 
1
2
Study Arm
-1
-.5
0
.5
1
composite

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to CT scan, and (2) to assess whether there are any potential safety concerns of the ProLung Test when 
used to evaluate patients with a positive CT scan. 
 
Subjects age 18 and above were recruited sequentially and randomly from those who were eligible and 
agreed to participate in the study, and were enrolled under an IRB-approved protocol (IRB 
# NA_00007789). All subjects presented with one or more suspicious radiological and/or clinical 
finding(s) for lung cancer, and all had undergone a CT scan of the lungs that indicated one or more non-
calcified nodules (NCN) or lung masses suspicious for lung cancer, or the subjects were scheduled to 
receive a CT scan of the lung within 14 days of enrollment.   
 
Forty-one patients were qualified to participate in the study.  The statistical approach used an optimal cut-
point methodology for classifying the two patient groups (malignant or non-malignant) based on the 
variables associated with the conductance measurements. The cut-point that provides the best test 
characteristics was considered the optimal cut-point.72 
xxxvi xxxvii
Results:  (Yung, 2012)xxxv 
 
.  Using this methodology for analyzing the ProLung’s bioconductance 
test data for the 41 patients resulted in the scatter plot in Figure 2.  As shown here, 11 of the 12 patients 
with benign lesions had composite scores that fell below the cut-point.  For the malignant cases, 26 of the 
29 had composite scores above the predetermined cut-off point with 3 falling below it. Computed bio-
conductance measurements discriminated between malignant lesions (29 primary lung cancers) and 
benign pathology (12) across a range of IPL sizes (0.8 cm and greater) with a sensitivity of 89.7% 
(positive predictive value 96.3%) and specificity of 91.7% (negative predictive value 78.5%). 
 
Conclusion.  This study found different bioconductance properties between cancerous and non-
cancerous lung lesions and provides encouraging results for continued development of the ProLung Test.   
 
 
 
Cut-off Point 

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Figure 2: Patient Bioconductance (ProLung) Test Results illustrate the composite scores for the 
benign patients (represented by the closed triangles) and the composite scores for the malignant cases 
(represented by the open squares) with reference to the Cutoff Point. 
 
3.0 
ProLung Unit: Technical Description 
The ProLung unit measures bioelectrical conductivity at multiple points on the skin’s surface. The probe 
introduces a low electrical current (too low to be felt) into the body and has adhesive electrodes that 
measure the current at other parts of the body. The ProLung unit is shown below in Figure 3 and has 
three functional components as illustrated in the schematic in Figure 4. The Measurement component 
includes both the hardware and software that obtain and record accurate and reproducible bioelectrical 
conductance measurements (data sets) of the thoracic cavity, referred to as the computerized 
bioconductance (ProLung) test output.  All of the software, hardware and unit configurations provided to 
the investigatory team, as illustrated below, are proprietary to the Sponsor.   
 
 
 
 
 
 
 
Figure 3. Fresh Medical Laboratories ProLung unit with probe (on left) and associated computer 
screen (on right) illustrating the Operator display. 
 
 
 
Figure 4.  ProLung Functional Components shown as a flow chart illustrating the sequence of function 
of the ProLung unit. 
 
3.1  Classifier Algorithm Design and Function 
With reference to Figure 4, above, following measurement, the characteristics of each subject’s data set 
are recorded and validated.  A ProLung data set or file consists of the measurements obtained at the 

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acquisition points for each subject.  The classifier Algorithm then analyzes the data set.  The algorithm 
processes specific properties of the data set to generate a report to aid the clinician in assessing the risk 
of malignancy associated with known lesions.     
3.2  Classification   
The ProLung report offers the test’s conclusions regarding patients with lesions; 1) high risk for 
malignancy or 2) low risk for malignancy.  With regulatory clearance, test results are to be used 
adjunctively with CT information and other risk factors to aid in the risk stratification of patients with 
indeterminate lung lesions that are suspicious for lung cancer. 
3.3 
Clinical Use  
The ProLung unit is designed for ease of operation, service and reliable performance. The ProLung unit 
conforms with all applicable safety standards and has been designated as a non significant risk device in 
its previous IRB reviews.  In preparation for investigational use, Fresh Medical Laboratories provides 
appropriate training. The ProLung provides screen prompts for essential subject information entry, and 
provides visual anatomic views to guide the operator through each measurement in the protocol 
sequence. The ProLung unit also provides visual and audible indications of measurement quality to the 
operator and the option of subject data measurement review by the operator. 
3.4 
ProLung Unit Physical Description  
The ProLung unit measures 13.5” wide by 13.15” deep by 3.15” high and integrates proprietary hardware 
and a Microsoft Windows™-based computer system within the enclosure. Typical computer operator 
interface and peripheral equipment is attached to the device including mouse, display and keyboard. The 
proprietary diaphoretic electrodes and computer driven data acquisition probe are connected by cords to 
easily connect to matching color coded ports on the left side of the device body. The data acquisition 
probe uses a unique, single-use tip that makes contact with the patient’s skin during the scan session.  
Marketed single-use kits will contain proprietary single-use tip and 6 (six) diaphoretic single-use adhesive 
reference electrodes. The patented probe design is critical to insure accurate and consistent 
measurements as described below.   A printer may be attached to the system. The ProLung unit is a 
portable self-contained device.  
3.5 
Safety Information 
The International Electrotechnical Commission (IEC) publishes the base standard 60601 that establishes 
limits for leakage current from a medical device at 150 microamps (0.000150 amps). The ProLung unit 
generates less than 10% of the allowable leakage current for medical devices. The instrument design 
includes industry approved isolation circuitry for use in clinical instrumentation.   
The ProLung unit delivers a safe current of <25 micro amps to measure bioconductance between 
reference electrodes placed on the patient’s back or hands and a computerized bioconductance probe 
placed sequentially at points on the skin surface. There is also safety information available from other 
currently marketed devices that introduce current into the body during usage.  To cite two modalities, 
transcutaneous electrical nerve stimulation (TENS) and percutaneous electrical nerve stimulation (PENS) 

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are used for acute post-operative or chronic intractable painxxxviii xxxix.  TENS is characterized by biphasic 
current and most TENS units produce a current of 1 to 80 microampere (mA), 9 V (average), 2 to 1000 
Hz, with a pulse width of 250 to 400 microseconds.  PENS uses needles as electrodes that are placed in 
soft tissues or muscles at dermatomal levels corresponding to the local pathology.  A 5
 
-Hz frequency with 
a pulse width of 0.5 mS is usually used.  Another FDA-approved device that delivers electrical stimulation 
is the ReliefBandR that is used to prevent or treat nausea and vomiting associated with motion sickness, 
pregnancy, or chemotherapyxl. Thus, the amount of electrical current that is associated with use of the 
ProLungis negligible and has not been nor is it expected to be associated with health risks or discomfort. 
The reviewing IRBs for all previous studies conducted for the Sponsor have determined that the ProLung 
Testposed a non-significant risk to study subjects.  In one ProLung study where 38 subjects were 
evaluated for safety, all completed the entire measurement session and none discontinued measurement 
due to discomfort or an adverse event (AE).  There were two AEs as follows: one AE consisted of 
transient minor pain at a specific measurement point which was determined by the Principal Investigator 
to be unrelated to measurement because the skin was irritated prior to measurement, and the second AE 
was moderate pain and a tingling paresthetics sensation at a few measurement points in an elderly 
subject (94 years of age) who had a documented history of severe, symptomatic osteoarthritis as well as 
pain with touch on subject’s skin.  The symptoms rapidly resolved after measurement completion.  The PI 
considered this AE to have a possible remote relationship to device use.  The subjects overwhelmingly 
reported satisfaction with the measurement protocol and have indicated without exception that they would 
agree to be tested again.  In the study with 22 normal subjects, none reported discomfort associated with 
use of the test.  In the latest study at Johns Hopkins with 55 patients, there were also no reported adverse 
events. 
The ProLung unit is manufactured under the Fresh Medical Laboratories ISO 13485 certified Quality 
Management System.  The ProLung unit has also completed IEC 60601-1 and IEC 60601-1-2 safety 
testing.   
 
 
 
 
 
 
 
 
 
 
 
 

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Appendix II: References 
 
 
                                                 
i Jemal Ahmedin et al., Cancer Statistics 2010, 60 Cancer Journal for Clinicians 5 (2010). 
 
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xxv Yoo DS.  The dielectric properties of cancerous tissues in a nude mouse xenograft model.  Biolectromagnetics 
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xxvii Aberg P, Nicander I, Hansson J et al. Skin cancer identification using multifrequency electrical impedance – a 
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spectroscopy: a prospective study.  Br J Obst. Gyn 2005; 112: 802-806 
 
xxxi Kimura S, Morimoto T, Uyama T, Monden Y, et al.  Application of electrical impedance analysis for diagnosis of 
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xxxii Kimura S, Morimoto T, Uyama T, Monden Y, et al.  Application of electrical impedance analysis for diagnosis of 
pulmonary mass Chest 1994;105;1679-1682.  Toso S, Piccoli A, Gusella M et al.  Altered tissue electric properties in 
lung cancer patients as detected by bioelectric impedance vector analysis. Nutrition 2000; 16(2): 120-124.  
Cherepenin V, Karpov A, Korjenevsky A et al. Preliminary static EIT images of the thorax in health and disease.  
Physiol Meas 2002; 23: 33-41 
 

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xxxiii Toso S, Piccoli A, Gusella M et al.  Altered tissue electric properties in lung cancer patients as detected by 
bioelectric impedance vector analysis. Nutrition 2000; 16(2): 120-124 
 
xxxiv Cherepenin V, Karpov A, Korjenevsky A et al. Preliminary static EIT images of the thorax in health and disease.  
Physiol Meas 2002; 23: 33-41 
 
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