A Prospective, Non-Randomized, Multi-Center Observational Study to Establish a Physical Baseline Profile for Individual Study Subjects Using Various Modalities and Identify Deviations Via Longitudinal Monitoring That May Develop Over Time
Conditions: Aging
Sponsor: Puer Research, LLC

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
1 
 
  
A Prospective, Non-Randomized, Multi–Center Observational Study to Establish a Physical Baseline 
Profile for Individual Study Subjects Using Various Modalities and Identify Deviations via 
Longitudinal Monitoring that May Develop Over Time and May be Relevant to Human Health and 
Longevity 
 
 
Protocol Short Title:   
PUER (“Previously Unrecognized Emerging Risks”) Life Clinical 
Study 
 
Identifying Number:    
 
PLI001  
 
Protocol Version and Date:    
Version v.2.0 06 October 2022 
 
Sponsor/Manufacturer:  
 
PUER Research, LLC 
 
 
8601 Dunwoody Place, Suite 304 
 
 
Atlanta, GA 30350 
 
 
United States 
 
Sponsor/Manufacturer Contact: 
Karl Blass 
 
 
 
 
 
PUER Research, LLC 
 
 
 
 
 
karl@puerlife.health 
 
 
 
 
 
404-277-7597 
 
Principal Investigator:  
 
Adam Friedlander, MD 
 
 
8601 Dunwoody Place, Suite 304 
 
 
Atlanta, GA 30350 
 
 
adam@puerlife.health 
 
 
 
 
 
678-792-8442 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
This document is CONFIDENTIAL and PROPRIETARY and shall not be disclosed without written 
consent from PUER Research, LLC. It is for use by the principal investigator and his/her designated 
representatives participating in this clinical investigation.  It should be held confidential and 
maintained in a secure location.  It should not be copied or made available for review by any 
unauthorized person or firm. 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
2 
 
PROTOCOL APPROVAL 
 
Protocol Number: PLI001 
 
A Prospective, Non-Randomized, Multi–Center Observational Study to Establish a Physical Baseline 
Profile for Individual Study Subjects Using Various Modalities and Identify Deviations via Longitudinal 
Monitoring that May Develop Over Time and May be Relevant to Human Health and Longevity.  
 
Protocol Version: Version 2.0 
 
Protocol Date: 06 October 2022 
 
This protocol has been read and approved by: 
Sponsor: 
 
________________________________________________________________________________ 
Name: Karl Blass  
 
 
 
Date: dd-mm-yyyy 
 
 
REVISION HISTORY  
Version 
Date 
Significant Revisions  
1.0 
08 August 2022 
Initial Version  
2.0 
06 October 2022 
Updated miscellaneous language and included language to 
exclude women with positive pregnancy test at any visit. 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
3 
 
STATEMENT OF INVESTIGATOR COMPLIANCE 
 
This protocol is a prospectively designed clinical study of the PUER Research, LLC  
I agree to: 
 
• 
Implement and conduct this study diligently and in strict compliance with the protocol, Good Clinical 
Practices (GCP) and all applicable laws and regulations. 
• 
Maintain all information supplied by PUER Research, LLC in confidence and, when this information 
is submitted to an Institutional Review Board (IRB), it will be submitted with a designation that the 
material is confidential. 
• 
Ensure that all persons assisting with the trial are adequately informed about the protocol and their trial-
related duties and functions. 
 
This document contains confidential information belonging to the Sponsor (PUER Research, LLC) and 
therefore, may not be disclosed to any other person or entity without the prior written permission of the 
Sponsor unless such disclosure is required by law or regulation. 
 
 
Investigator Signature  
 
I have read and understand the contents of the clinical protocol including this Statement of Investigator 
Compliance.  I agree to follow and abide by the guidelines set forth in this document.  
 
Adam Friedlander, MD 
Principal Investigator Name  
 
 
 
 
 
 
 
 
 
 
 
 
  
Principal Investigator Signature   
 
 
 
Date (dd-mmm-yyyy) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
4 
 
 
Protocol Synopsis 
 
Study Title: A Prospective, Non-Randomized, Multi–Center Observational Study to Establish a Physical 
Baseline Profile for Individual Study Subjects Using Various Modalities and Identify Deviations via 
Longitudinal Monitoring that May Develop Over Time and May be Relevant to Human Health and 
Longevity.  
 
Short Title 
PUER Research Clinical Study 
Study Type 
This is a minimal-risk, prospective, non-randomized, multi-center population based 
observational study to establish a physical baseline profile for individual study 
subjects using various modalities and identify deviations through longitudinal 
monitoring that may develop over time and may be relevant to human health and 
healthy longevity.  
Study 
Description  
The three key elements of the PUER Research protocol include (1) Medical history 
and physical exam (PE), (2) molecular and laboratory profiling and (3) non-invasive 
imaging and wearables/ “quantified self” measurements. The medical history and 
PE will consist of a comprehensive medical history and physical examination, 
administered by a trained Research Associate. Laboratory assessments will be 
conducted using peripheral blood samples, and potentially urine, stool or saliva 
samples. Non-invasive imaging assessment will consist of magnetic resonance 
imaging (MRI) and computerized tomography (CT) scans. Other functional 
assessments, including respiratory, cardiac and other functions, as well as self-
quantifiable assessments via wearables will also be conducted.  
  
The study will result in longitudinal real world data collection to inform if any 
modalities employed here can inform of baseline deviations for individuals. 
Researchers at study sites will not be blinded to the data being collected during this 
study. 
Study Sites 
This study will be conducted in up to 20 study sites in the United States.  
Study 
Population 
The study will be conducted in up to 10,000 participants. 
Study Duration  
72 months  
Enrollment 
Duration 
36 months   
Objectives 
• 
The primary objective is to establish a baseline profile for individual healthy 
study subjects using various modalities and monitor them longitudinally to 
identify deviations that develop over time and may be relevant to human health 
and longevity The secondary objective is to identify the modality that has the 
highest impact on the identification of deviations from baseline profile. 
Adverse 
Event 
(AE) Monitoring 
Adverse events associated with the study procedures, of venipuncture, non-invasive 
imaging and functional tests will be reported, although they are expected to be 
limited due to the nature of these common procedures. 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
5 
 
Inclusion 
Criteria 
To participate in the study, patients must meet the following criteria;  
1. Male or non-pregnant female; age 18 to 90. 
2. Absence or presence of any medical history or any signs or symptoms of 
any disease. 
3. Women of childbearing potential (WOCBP) must have a negative urine 
pregnancy test (UPT) at all visits.  
 
Exclusion 
Criteria 
Patients who meet any of the following criteria will be excluded from study 
participation;  
1. Unwillingness or inability to participate in the study 
2. Unwillingness or inability to provide written Informed Consent Form 
3. WOCBP with positive pregnancy test at enrollment or at any visit 
Study Schedule 
 
Event 
Baseline 
Visit 
6-
Month 
Visit ± 
1month 
12-
Month 
Visit ± 
1month 
18-
Month 
Visit ± 
1month 
24-
Month 
Visit ± 
1month 
36-
Month 
Visit 
± 
1month 
Informed Consent 
X 
 
 
 
 
 
Review of 
Inclusion/Exclusion 
Criteria 
X 
 
 
 
 
 
Medical history 
and physical exam 
X 
X 
X 
X 
X 
X 
UPT1 for WOCBP2 
X 
X 
X 
X 
X 
X 
Whole Genome 
Sequencing 
Analysis 
X 
 
 
 
 
 
Biochemical/Omics 
Analysis 
X 
X 
X 
X 
X 
X 
Non-Invasive 
Imaging3 
X 
X 
X 
X 
X 
X 
Functional 
Assessments (as 
needed) 
 
X 
X 
X 
X 
X 
Wearables and 
Quantified Self 
Assessments (as 
needed) 
 
X 
X 
X 
X 
X 
Adverse Event 
Monitoring 
X 
X 
X 
X 
X 
X 
1. UPTs must have a minimum sensitivity of 25 mIU ß-hCG/mL. 
2. WOCBP include any female who has experienced menarche or is 10 years of age or older and who has not 
undergone successful surgical sterilization (hysterectomy, bilateral tubal ligation, or bilateral 
oophorectomy) or is not postmenopausal (defined as amenorrhea >12 consecutive months in women 50 
years of age and older). 
3. See Section 3.3.2 for detailed schedule of imaging 
 
 
 
 
 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
6 
 
 
 
ABBREVIATIONS 
 
AE 
Adverse Event 
CFR 
Code of Federal Regulations 
CRF 
Case Report Form 
CT 
Computed Tomography 
DNA 
Deoxyribonucleic Acid 
ECG 
Electrocardiogram 
EDC 
Electronic Data Capture 
EEG 
Electroencephalogram 
EMG 
Electromyography 
FDA 
Food and Drug Administration 
GBCM 
Gadolinium-Based Contrast Media 
GCP 
Good Clinical Practice 
GINA 
Genetic Information Nondiscrimination Act 
HR 
Heart Rate 
HRV 
Heart Rate Variability 
IRB 
Institutional Review Board 
miR 
Micro-RNA 
mL 
Milliliter 
mRNA 
Messenger RNA 
MRI 
Magnetic Resonance Imaging 
NNT 
PE 
Number Needed to Treat 
Physical Examination 
PHI 
Protected Health Information 
PMI 
Precision Medicine Initiative 
RNA 
Ribonucleic Acid 
SAE 
UPT 
Serious Adverse Event 
Urine Pregnancy Test 
US 
United States 
WGS 
WOCBP 
Whole Genome Sequencing 
Women of Childbearing Potential 
 
 
 
 
 
 
 
 
 
 
 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
7 
 
 
Table of Contents 
 
PROTOCOL SYNOPSIS ........................................................................................................................... 4 
ABBREVIATIONS ..................................................................................................................................... 6 
TABLE OF CONTENTS ........................................................................................................................... 7 
1. 
INTRODUCTION ............................................................................................................................... 9 
1.1. 
BACKGROUND ................................................................................................................................ 9 
1.2. 
RATIONALE ..................................................................................................................................... 9 
2. 
OBJECTIVES AND ENDPOINTS .................................................................................................. 12 
2.1. 
MAIN OBJECTIVES: ....................................................................................................................... 12 
2.2. 
MAIN ENDPOINTS: ........................................................................................................................ 12 
2.3. 
EXPLORATORY OBJECTIVES: ........................................................................................................ 12 
3. 
STUDY DESIGN ............................................................................................................................... 14 
3.1. 
STUDY SCHEMA ............................................................................................................................ 14 
3.2. 
GENERAL SELECTION CRITERIA ................................................................................................... 14 
3.2.1. 
Inclusion Criteria .................................................................................................................. 14 
3.2.2. 
Exclusion Criteria ................................................................................................................. 14 
3.3. 
STUDY PROCEDURES .................................................................................................................... 15 
3.3.1. 
Study Design .......................................................................................................................... 15 
3.3.2. 
Methodology .......................................................................................................................... 15 
4. 
BENEFIT/RISK ASSESSMENT ..................................................................................................... 18 
4.1. 
POTENTIAL RISKS ......................................................................................................................... 18 
4.2. 
POTENTIAL BENEFITS ................................................................................................................... 19 
4.3. 
OVERALL BENEFIT/RISK RATIO ................................................................................................... 19 
5. 
ADVERSE EVENTS ......................................................................................................................... 20 
5.1. 
DEFINITIONS ................................................................................................................................. 20 
5.2. 
PREGNANCY .................................................................................................................................. 20 
5.3. 
REPORTING ................................................................................................................................... 20 
5.4. 
SAFETY ......................................................................................................................................... 21 
6. 
DATA MANAGEMENT ................................................................................................................... 22 
6.1. 
CASE REPORT FORMS ................................................................................................................... 22 
6.2. 
DATABASE .................................................................................................................................... 22 
6.3. 
SAFEGUARDING OF GENOMIC DATA ............................................................................................ 22 
6.4. 
QUALITY CONTROL ...................................................................................................................... 23 
7. 
STATISTICAL CONSIDERATIONS ............................................................................................. 24 
8. 
ETHICAL, REGULATORY, AND ADMINISTRATIVE CONSIDERATIONS ....................... 25 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
8 
 
8.1. 
INFORMED CONSENT .................................................................................................................... 25 
8.2. 
CONFIDENTIALITY ........................................................................................................................ 25 
8.3. 
INSTITUTIONAL REVIEW ............................................................................................................... 25 
8.4. 
PROTOCOL INTERPRETATION AND COMPLIANCE ......................................................................... 25 
8.5. 
COMPLETION OF CASE REPORT FORMS (CRFS) ........................................................................... 25 
8.6. 
MAINTENANCE OF STUDY DOCUMENTATION .............................................................................. 26 
8.7. 
PUBLICATION POLICY ................................................................................................................... 26 
8.8. 
FINANCING AND INSURANCE ........................................................................................................ 26 
8.9. 
FINAL STUDY REPORT .................................................................................................................. 26 
8.10. 
RECORD RETENTION ................................................................................................................. 26 
9. 
REFERENCES .................................................................................................................................. 27 
 
 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
9 
 
 
1. Introduction 
 
1.1. 
 Background 
 
Scientific and medical progress has been exponential over the past decades, yet the implementation of 
new technologies into every-day clinical practice can lag by 5-10 years, thus preventing a large portion 
of the population from benefiting from those advances during their meaningful lifetime (Glasziou & 
Haynes, 2005). Earlier detection leads to earlier intervention where needed and accordingly, deploying 
novel technologies is the cornerstone of avoiding undesirable outcomes. However, the path and timeline 
of discovery, development, regulatory approval, and commercialization can delay implementation of 
potentially life-saving technologies by 5-10 years. 
 
There have been tremendous advances in the understanding of health, and longevity over the past 
decades. The early 2000s marked the ushering in of the genomic era, with the sequencing of the human 
genome (Guttmacher & Collins, 2003). Since that time, the required time and cost of genome 
sequencing has dropped dramatically, and the actionability of sequencing the entire human genome has 
increased exponentially (Hayden, 2014) (Scaria, 2014) (Dorschner, et al., 2013) (Amendola, et al., 
2015) (Goldstein, 2013) (Bromberg, 2013). Beyond DNA sequencing, other “omics” analyses such as 
DNA methylation, RNA sequencing, proteomics, metabolomics and lipidomics technologies have 
matured to the point of significant reproducibility, providing important biological insights. 
Conventional biochemical laboratory technologies have improved and matured and there are a large 
number of diagnostics laboratory tests available in the commercial setting.  
 
Similarly, non-invasive imaging techniques, such as magnetic resonance imaging (MRI) and computed 
tomography (CT) modalities have continued to evolve, using artificial intelligence- and computer-aided 
design-based analytical methods to improve the quality of diagnostic images and enhance clinical care 
(Uppot, et al.,, 2018) (Onal, et al., 2014) (Zhu, et al., 2018) (Anthimopoulos, et al., 2016)  (Li, et al., 
2008) (Arnoldi, et al., 2010) (Lemaitre, et al., 2015) (Yang, et al., 2014) 
 
The above techniques, as well as others, can now be applied both in healthy individuals, as well as in 
individuals exhibiting certain deviations from a healthy baseline profile. However, the impact of 
deploying a large suite of genomic, biochemical, and non-invasive imaging modalities on human health 
and longevity is unknown. Understanding such impact of the combination of such modalities, as well 
as the relative contribution of individual modalities, represents a leap forward in the understanding of 
human health and healthy longevity. 
 
1.2. 
Rationale 
 
The fundamental basis of the PUER (“Previously Unrecognized Evidence of Risk”) study is based in 
the central dogma of biology (Crick, 1970). The central dogma of biology stipulates that human health 
and longevity are determined in the context of the interactions of the human genome (DNA) and the 
environment, and these interactions will determine the transcription of DNA to RNA, the translation of 
RNA to proteins, and such structural and functional proteins will determine the complex molecular 
milieu in an individual, leading to quantifiable phenotypes of health and disease. The human genome 
(DNA) contains approximately 6.4 billion nucleotides organized into ~25,000 genes, transcribed into 
approximately 100,000 different RNA transcripts and ~25,000-28,000 transcribed gene constructs that 
are translated into about 20,000 human proteins. These proteins make up human cells and organs and 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
10 
 
regulate the intracellular and extracellular milieu of the human body, containing over 10,000 small 
molecules and lipid/phospholipid species.  
 
Technologies exist today to measure the majority of these biochemical and “omics” components. Whole 
genome sequencing (WGS) technologies are now able to decipher all 6.4 billion nucleotides in each 
individual (“wet lab component”), and cutting-edge bioinformatics pipelines can tie individual genetic 
variants to disease susceptibility, pharmacogenomics, and disease carrier status (“dry lab component”) 
(Netto & Kaul, 2019). Both messenger RNA (mRNA) and micro-RNA (miR) can be sequenced from 
blood-based, peripheral circulating cells and such gene expression signatures have already been 
translated to clinically approved diagnostic blood tests (Rosenberg, et al., 2010). A number of different 
techniques are available for proteomics profiling based on mass spectrometry (Han, et al.,  2008) and 
other techniques, such as aptamers, somamers (Gold, 2010) and dual-antibody approaches (Assarsson, 
et al., 2014). Such proteomics signatures have also been turned into diagnostic signatures (Ganz, et al., 
2016). Metabolomics profiling is now also a reality, using mass spectrometry (Yet, et al., 2016), NMR-
technologies (Fest, et al., 2019) and other techniques (Bennuru, et al., 2017). Similar to metabolomics, 
lipidomics profiling is also available today (David, et al., 2022). Many commercial laboratories have 
developed Food and Drug Administration (FDA)-cleared diagnostic and screening tests that can be 
applied in everyday clinical practice.  
 
While non-invasive imaging modalities, such as CT and MRI have been around for several decades, 
artificial intelligence-based post-processing and interpretation approaches are transforming the imaging 
field with the potential to radically impact clinical care (Uppot, et al., 2018).  
 
Yet another important concept is the concept of the “N-of-1” clinical study (Schork, 2015). The 
traditional path in science and medicine has been a population-based approach, where large populations 
were compared across different timepoints. Accordingly, most nutraceutical, pharmaceutical and 
medical interventions have been developed for populations, with varying degree of efficacy in 
individuals. The most important and most impactful drugs in the history of medicine have been limited 
on an individual basis. The “number needed to treat” (NNT) is the number of people who need to 
receive the drug (or other intervention) in order for just one person to receive a benefit, can be an 
important number to consider. For reference, an NNT of 5 or less (≤5) is probably associated with a 
meaningful health benefit for an individual, while an NNT of 15 or more (≥15) is quite certain to be 
associated with, at most, a small net health benefit (Chong, et al., 2006). The NNT for the most 
successful drugs in history is quite high. For simvastatin, the NNT is approximately 81, which is a 
staggeringly high number (“Design and baseline” 1993) (“Randomized trial” 1994). Precision medicine 
and “N-of-1 studies” may provide an opportunity to significantly increase the efficacy of certain 
nutraceuticals, pharmaceuticals, and other medical interventions.  
 
In the context of laboratory evaluation and the evaluation of certain biochemicals and laboratory 
measurements, the concept of the “N-of-1 study” stipulates that rather than using population-based cut-
points for laboratory values and applying them to individuals, we should use an individual as their own 
control over time. If we are able to monitor a large suite of biochemicals and laboratory measurements 
in an individual over time, this will establish their own baseline, and any significant deviation from 
their own baseline may signal the need for further evaluation and testing by the study subject’s health 
care provider. This concept leads to the approach of longitudinal monitoring and surveillance over time.  
 
Fortunately, the concept of individualized, or “precision medicine” has been gaining traction even at 
the level of the United States (U.S.) federal government. The Precision Medicine Initiative (PMI) was 
unveiled at the State of the Union Address on January 30, 2015 and had an initial investment of $215M 
from the 2016 federal budget. The distilled goal of the program is “to accelerate biomedical discoveries 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
11 
 
and provide clinicians with new tools, knowledge, and therapies to select which treatments will work 
best for which patients.”  Simply, the PMI seeks to shift the healthcare paradigm from a one-size-fits-
all approach to one which individualizes treatment plans based on individual differences in genetics, 
lifestyle, and environment. The original language of the initiative specifically mentions metabolomics 
and the microbiome, signaling the acceptance and promotion of cutting-edge health technology and 
research, beyond what is currently considered standard of care. Success in this area exists, most 
recognizably in the field of oncology, but the next steps into this frontier require the translation and 
broadening of current concepts to include more patients and every field of medicine. To do this, public 
and private collaboration will be required to leverage advances in genomics, big data, and health 
information technology, specifically regarding health data portability, for collaboration between 
patients and multidisciplinary healthcare teams. The PMI calls on millions of Americans to contribute 
health data to “catalyze a new era of data-based and more precise medical treatment.”  Participants will 
have granular control of the privacy of the data they share and may withdraw consent for any component 
at any time, however the goal will be the creation of secure, trustworthy platforms which will engender 
confidence in sharing deidentified data on a longitudinal, ongoing basis, allowing physician scientists 
to collect and analyze that data over the course of millions of lifetimes. The PMI’s All of US research 
program is already the largest longitudinal study in the history of the United States.  
 
The initial objectives of the PMI focused on strengthening existing areas of success, while broadening 
federal support and soliciting private support for both existing and new initiatives, with definitive 
commitment to patient engagement, privacy, and data security.  The language of the PMI discusses the 
creation of a “cancer knowledge network” via funding to the National Cancer Institute for the design 
and testing of tailored treatments for cancer based on the genetics of patients and specific cancers. Next, 
the initiative calls for multi-departmental collaboration on the creation of a voluntary national research 
project via the National Institutes of Health, The Department of Health and Human Services, The 
Department of Veterans Affairs, The Department of Defense, and other relevant stakeholders, with the 
goal of enrolling over a million diverse individuals. Through this project, participants will contribute a 
wide variety of data sources “including medical records; profiles of the patient’s genes, metabolites 
(chemical makeup), and microorganisms in and on the body; environmental and lifestyle data; patient-
generated information; and personal device and sensor data.” Radiology studies are included in this 
complete digital health profile, as well, as is all other available historical data, and all new data collected 
via the participating programs. To date, radiology studies have played a key role in the phenotyping 
that goes hand-in-hand with advanced genomics exploration. The amount of information in a single 
individual’s digital health profile can be massive, and for that reason, the big data component of the 
PMI is critical to the initiative’s success. As to privacy and data security, the PMI definitively commits 
to soliciting input from federal, public, and private entities to identify and address legal and technical 
issues surrounding privacy and data security, while the FDA will focus on “regulatory modernization” 
with specific attention to genetic testing. Lastly, the PMI emphasizes the importance of forging strong 
public-private partnerships for infrastructure development and patient enrollment, with attention to 
patient engagement and empowerment throughout each individual’s experience over their lifetimes. 
 
Accordingly, the objective of the PUER study is to assess the impact of longitudinal, comprehensive 
evaluation based on certain currently available modalities in a large, unselected population of 
individuals.  
 
If our study demonstrates that comprehensive baseline and longitudinal assessment employing these 
various modalities  identifies previously unknown deviations from a study subject’s baseline, this could 
inform lifestyle changes and other potential interventions to maintain a healthier self. Longitudinal 
testing at frequent intervals, such as every 6 months, is critical, as metabolomic and proteomic changes 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
12 
 
occur on the order of hours and days, and diseases such as pancreatic cancer can progress rapidly within 
a 4 to 6 month interval. 
 
 
2. Objectives and Endpoints 
 
2.1. 
Main Objectives: 
 
 
To establish a baseline profile for individual study subjects and monitor them 
longitudinally to identify deviations that develop over time and may be relevant to 
human health and healthy longevity.  
 
To identify the modality that has the highest impact on the identification of 
deviations from baseline profile. 
 
 
2.2. 
Main Endpoints: 
 
 
 
The number of new deviations from a study subject’s baseline profile 12 months 
after enrollment that were previously not known at time of enrollment 
 
 
The number of new deviations not known at the time of enrollment, at the 
following timepoints after enrollment: 
• 
6 months after enrollment 
• 
18 months after enrollment 
• 
24 months after enrollment 
 
 
The number of new deviations identified by each employed modality 
 
 
2.3.  
Exploratory Objectives: 
 
 
To assess the impact of introducing leading-edge scientific methods on the number 
of medications and/or dietary supplements taken by individuals 
 
 
To assess the impact of leading-edge scientific methods on the number of 
unscheduled health care visits 
 
 
2.4.  
Exploratory Endpoints 
 
 
The number and change in the number of medications taken by an individual at 6 
months, 12 months, 18 months, and 24 months after enrollment 
 
 
The number and change in the number of dietary supplements taken by an 
individual at 6 months, 12 months, 18 months, and 24 months after enrollment 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
13 
 
 
 
Number of unscheduled, urgent care/emergency department visits within 24 
months and 36 months after enrollment 
 
 
Number of unscheduled hospital admissions within 24 months and 36 months after 
enrollment 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
14 
 
 
3. Study Design 
 
This is a prospective, population-based observational study. Participants may be recruited from clinical 
sites that are actively participating in the PUER Research comprehensive evaluation model. Participants 
and all participating health care providers are unblinded.  
 
 
3.1. 
Study Schema  
 
 
 
 
Figure 1. Overall Study Schema. The Baseline Visit includes a Medical history and physical examination (PE) Genomic 
Analysis, Biochemical/”Omics” Analysis and Non-Invasive Imaging. Subsequent visits at 6 months, 12 months, 18 months, 
24 months and 36 months include slightly modified and targeted Medical history and PE, Biochemical/”Omics” Analysis 
and Non-Invasive Imaging, based on prior findings. Functional Assessments and Wearables and Quantified Self Assessments 
are included at the 6 months, 12 months, 18 months, 24 months and 36 months visits as determined by the Medical history 
and PE at the baseline visit. 
3.2. 
General Selection Criteria 
 
To be included in the study, participants must meet the following inclusion criteria and none of the 
exclusion criteria. 
 
3.2.1. Inclusion Criteria 
• 
Male or non-pregnant female; age 18 to 90 
• 
Absence or presence of any medical history or any signs or symptoms of any 
disease 
• 
Women of childbearing potential (WOCBP) must have a negative urine pregnancy 
test (UPT) at all visits.  
 
3.2.2. Exclusion Criteria 
• 
Unwillingness or inability to participate in the study 
• 
Unwillingness or inability to provide written Informed Consent Form 
• 
WOCBP with positive pregnancy test at enrollment or at any visit 
 
3.2.3 Participant withdrawal criteria 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
15 
 
A participant may be withdrawn from the study prior to completion for any of the following 
reasons: 
 
• Adverse Event (AE) 
• Death 
• Physician decision 
• Pregnancy at any visit 
• Protocol deviation 
• Other (e.g., any reason that may affect the outcome of the study or safety of 
participants) 
• Participant no longer wants to participate (without penalty) 
 
When a participant is withdrawn from the study for an AE (as defined in Section 5), when possible, the 
participant should be followed until resolution or stabilization of the AE. 
 
 
3.3. 
Study Procedures 
 
3.3.1. Study Design  
 
• 
This is a prospective, population-based observational study. Participants may be 
recruited from clinical sites that are actively participating in the PUER ResearchTM 
Comprehensive Evaluation protocol. Participants and all participating health care 
providers are unblinded. 
• 
The three key elements of the PUER Research protocol include (1) Medical history 
and PE, (2) molecular and laboratory profiling and (3) non-invasive imaging, with 
the option of adding functional assessments and wearables/quantified self-
assessments, as determined by the Medical history and PE.  
3.3.2. Methodology 
 
Each of the visits include a combination of Medical history and PE, biochemical and 
“omics” assessments, non-invasive imaging assessments and optional functional 
assessments and wearables and quantified self-assessments. 
 
 
Medical history and PEThe Medical history and PE consists of a comprehensive 
“History and Physical Examination”, administered by a trained Research 
Associate. The elements of the history and physical examination are recorded in 
the Case Report Form (CRF). 
• 
Pregnancy Testing 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
16 
 
Pregnancy testing consists of a UPT for all WOCBP1. The results must be 
negative for the subject to be able to participate in each visit. 
 
• 
Genomic Analysis 
 Whole genome sequencing (WGS) analysis (“DNA”) 
  
• 
Biochemical and “Omics” Assessments 
Biochemical and “omics” assessments consist of the following elements, using 
peripheral blood samples, and potentially urine, stool or saliva samples.  
 Whole genome methylation analysis (“DNA methylation”) 
 Whole transcriptome sequencing from circulating mononuclear cells (mRNA 
and miR) 
 Proteomics profiling 
 Metabolomics profiling 
 Conventional biomarker analysis 
 Urine analysis and profiling 
 
• 
Non-Invasive Imaging Assessments 
 Non-invasive imaging assessment consists of the following elements, as directed, 
and determined by the Medical history and PE and biochemical and “omics” 
assessment, as described above. Imaging will follow the schedule in the table 
below, with an “x” indicating all participants and an “[x]” indicating imaging 
needed for risk-specific participants. Any deviation from the average population 
may trigger a dedicated organ specific MRI. Imaging may also be done at 18-, 24- 
and/or 36-month visits, depending on Medical history and PE at each visit, 
following the schedule at 6 or 12 months. 
 
Imaging Procedure 
Baseline 
6 mos 
12 mos 
CT of head 
x 
 
 
CT Angiogram of head/neck/face/sinus/orbits 
x 
 
 
MRI of brain with and without contrast 
x 
 
x 
MR Angiogram of head/neck time of flight 
 
 
x 
MRI of brain without contrast 
 
[x] 
 
CT of chest, abdomen, and pelvis - mixed A/V 
x 
[x] 
[x] 
CT of chest, abdomen, and pelvis - venous only 
 
[x] 
x 
MRI of spine without contrast 
(cervical/thoracic/lumbar) 
x 
 
x 
 
1 WOCBP include any female who has experienced menarche or is 10 years of age or older and who has not undergone 
successful surgical sterilization (hysterectomy, bilateral tubal ligation, or bilateral oophorectomy) or is not 
postmenopausal (defined as amenorrhea >12 consecutive months in women 50 years of age and older). 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
17 
 
X-rays of extremities 
x 
[x] 
x 
Ultrasound (reproductive organs/bladder) 
x 
[x] 
x 
 
• 
Functional Assessments (determined by the Medical history and PE, described 
above) 
 VO2-max functional testing (maximum oxygen consumption) 
 Non-invasive endothelial function testing 
 Electroencephalogram (EEG) 
 Electromyography (EMG) 
 Ophthalmologic examination for vision preservation and other pathology 
 Optional: sleep study 
 
• 
Wearables and Quantified Self Assessments (determined by the Medical history 
and PE, described above) 
 Ambulatory Electrocardiogram (ECG) monitoring (Apple Watch, ePatch or 
similar) 
 Continuous glucose monitoring 
 Oura ring (may obviate the need for formal sleep study), or similar product(s) 
measuring heart rate (HR), heart rate variation (HRV), sleep duration, sleep 
stage 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
18 
 
 
4. Benefit/Risk Assessment 
 
4.1. 
Potential Risks 
 
The potential risks of the PUER study are minimal. The PUER study does not prescribe any 
pharmaceutical or medical device-based interventions. Information collected from the genomic 
analysis, comprehensive “omics” analysis, non-invasive imaging and functional assessment will be 
shared with the individual and, if desired, with the health care provider of choice of the individual. Any 
potential treatment decision of further diagnostic evaluation-related decision will be made by the 
individual and their chosen health care provider.  
 
 
Accordingly, any potential risks of the PUER study are only related to the comprehensive evaluation. 
For comprehensive biochemical and “omics” profiling, individuals will undergo blood draw through 
venipuncture. The risks of blood draw and venipuncture are minimal and can be mitigated. AEs that 
may be associated with venipuncture and that must be included in the informed consent include: 
• 
Pain 
• 
Bruising 
• 
Bleeding at the puncture site 
• 
Fainting 
• 
Inflammation of the vein 
 
Individuals will also undergo MRI. MRI is a safe, non-radiation-based modality and when 
contraindication to MRI are adequately assessed, MRI examination does not pose any risk to the 
participants. Standard screening for contraindications to MRI examination will be performed at the 
imaging center, using the centers’ policies and procedures for screening to identify contraindications to 
MRI examinations. Gadolinium-based contrast agents may be administered in connection with the MRI 
examination. Gadolinium-based contrast agents have been associated with potential risks, including 
rare anaphylactic reactions and necrotizing systemic fibrosis (American College of Radiology, 2021). 
 
 
The use of gadolinium contrast agents may be associated with anaphylactic reaction in 0.001% to 0.01% 
of cases, and gadolinium contrast agents have also been associated with nephrogenic systemic fibrosis 
(NSF) in patients with compromised renal function. Based on the 2021 ACR Manual on Contrast 
Media, any adverse event rate with gadolinium-based contrast media (“GBCM”) ranges from 0.07% to 
2.4%. Most reactions are mild and physiologic. Allergic-like reactions are uncommon and vary in 
frequency from 0.004% to 0.7%. Severe life-threatening anaphylactic reactions are exceedingly rare 
(0.001% to 0.01%). 
 
Individuals will also undergo CT. CT is a safe imaging method that includes exposure to ionizing 
radiation with or without contrast agents. The contrast is either given orally, rectally or intravenously. 
This contrast material can rarely cause medical problems or allergic reactions. Most of these reactions 
are mild and result in a rash or itchiness. In rare instances, an allergic reaction can be serious, even life-
threatening. 
 
During an abdominal ultrasound exam, a trained technician will glide a small device over the 
participant’s abdomen to take pictures of the tissue under the skin. There are no known side effects 
from taking part in an abdominal ultrasound exam. Ultrasound tests do not involve radiation and they 
do not penetrate the skin. 
 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
19 
 
Due to the risk of imaging radiation exposure to a fetus, WOCBP must not be pregnant at any visit. 
 
Individuals will also undergo x-ray imaging. X-ray is an imaging method that includes exposure to a 
low amount of electromagnetic radiation to take pictures of structures in the body, such as bones. 
 
4.2. 
Potential Benefits 
 
Participants may not derive any benefit at all from participating in the study. At the same time, the 
extensive genetic, biochemical, and non-invasive imaging evaluation may uncover unknown genetic 
susceptibilities and/or early identification of variance and deviations from a study subject’s baseline 
profile that may be worthy of further evaluation and testing by the study subject’s primary care 
physician, providing some potential benefits to participants.  
 
 
4.3. 
Overall Benefit/Risk Ratio 
 
Given that potential risks are minimal or negligible as described above, and given potential benefits as 
described above, the overall benefit/risk ratio favors participation in the study.  
 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
20 
 
 
5. Adverse Events 
 
5.1. 
Definitions 
 
An Adverse Event (AE) is a noxious and unintended event observed in, or reported by, a participant 
who is participating in (or has participated in) a clinical study, and/or received study medication, an 
experimental device, or study-related procedure.  The event may be related temporally either to 
immediate or long-term use of a drug, device, or procedure.  It may not necessarily be caused by a drug, 
device, or procedure.  Any event meeting these criteria shall be considered an adverse event, regardless 
of whether or not it is considered study related.  A pre-existing condition that worsens during the trial 
is considered an AE. 
 
The following events will be considered as adverse events: 
• 
Event directly related to study procedure 
• 
Unintended event as a result of non-standard of care study tests, medications or 
procedures 
 
A Serious Adverse Event (SAE) is defined as one that is fatal or immediately life-threatening, severely 
or permanently disabling, requires or prolongs in patient hospitalization, a congenital anomaly, cancer 
or overdose (intentional or accidental); or suggests a significant hazard, contraindication, side effect, 
or precaution.  All SAEs will be followed by the investigator through trial end until resolution or until 
reaching a clinically stable status per the investigator’s discretion. 
 
The following adverse events will be handled as serious even if they do not meet the above criteria for 
designation as serious: 
• 
Death 
• 
Congenital abnormality 
• 
Hospitalization 
• 
Life threatening illness 
• 
Intervention to prevent permanent damage 
 
5.2. 
Pregnancy 
 
WOCBP (see Protocol Synopsis for definition of WOCBP) must have a negative UPT prior to study 
enrollment and at each study visit. Prior to study enrollment, WOCBP must be advised of the 
importance of avoiding pregnancy at each visit. The subject must sign an informed consent form 
documenting this discussion. 
 
WOCBP should be instructed that if they are pregnant, they will not be permitted to be enrolled and are 
subject to withdrawal from the study with a positive UPT at any visit, as per Section 3.2.3. 
 
 
5.3. 
Reporting 
 
The PUER study does not prescribe any pharmaceutical or medical device-based intervention. Any AEs 
are associated with blood draw/venipuncture and MRI examinations that may or may not include 
Gadolinium-based contrast agents will be reported. 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
21 
 
 
Each patient will be observed and queried in a nonspecific fashion at each contact during the study for 
any new or continuing symptoms since the last contact. All adverse events will be reported on the 
appropriate CRF.  Details will include the type of event, date of onset, duration, intensity, causality 
relationship to the study procedure (if applicable), and outcome.  Wherever possible, a diagnosis rather 
than symptom(s) will be reported. 
 
If an AE should occur, every attempt will be made to obtain as much information as possible about 
event evaluation and outcome.  Documents of this follow-up will be maintained with the patient’s study 
records.   
 
If a SAE occurs, the participation in the study will be interrupted or discontinued at the investigator's 
discretion.   
 
All protocol deviations will be reported to the WCG IRB.   
 
All SAEs will be reported immediately to the WCG IRB and the FDA (if applicable). 
 
Endpoints will be adjudicated by the Principal Investigator.  A written report detailing the endpoint 
adjudication will be provided by the Principal Investigator. 
 
5.4. 
Safety 
 
This study will be conducted in accordance with the principles of Good Clinical Practice (GCP) as 
outlined in the Declaration of Helsinki and the International Conference on Harmonization. 
 
The principal investigator of the study will review study related events for adjudication.  
 
All efforts will take place to ensure patient safety. Each patient will be monitored for safety throughout 
the trial utilizing clinical evaluations and laboratory markers.  Laboratory markers and/or clinical 
evaluations that are out of normal range will be recorded as adverse events and reviewed with the 
investigator.  

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
22 
 
 
6. Data Management  
 
Following completion of the consenting process (including informed consent and Authorization for 
Use/Disclosure of protected health information (PHI)) and it has been determined that the patient meets 
all of the inclusion and none of the exclusion criteria, enrollment will occur, and data collection will 
commence. 
 
A secure Electronic Data Capture (EDC) and data management system will be used for entry, storage, 
review, and management of study data. The system will be compliant with applicable GCP and 
regulatory requirements. Investigators are responsible for accurate completion and timely submission 
of the data collected during the study. Sites will be trained in the use of the EDC system for entering 
study data and uploading supporting documents and will be given access for this purpose.  
 
Quality assurance procedures will be established to ensure that complete, accurate and timely data are 
submitted, protocol requirements are followed, and that AEs are correctly reported. The management 
and retention of these data will be compliant with applicable regulatory requirements. 
 
6.1. 
Case Report Forms 
 
All required data for this study will be collected on standardized paper CRFs. The research staff will 
review completed CRFs to attest that all data entered on them are complete and accurate. 
 
Medical history, demographics, hospital, office, and research records for any visit (including any notes, 
reports, test results, and lab reports from the clinical evaluation, biochemical and “omics” assessment, 
non-invasive imaging, functional assessments, and wearable/quantitative self-assessments) are 
considered source documentation and will be collected and reviewed to confirm clinical events and 
may be utilized for data analysis. Data will be collected on all patients and entered into a CRF.  
 
The Investigator or designee will be responsible for completing, in a timely manner, the CRF for each 
participant related to previously reported or newly detected AEs. And end of study withdrawal form 
will also be included as part of the CRF.  
 
6.2. 
Database 
 
Data from the CRF will be transcribed to the EDC system. Data collected for omics and imaging will 
be retained with 3rd parties conducting these analyses.   
 
6.3. 
 Safeguarding of Genomic Data  
 
Some blood samples will be used for genetic testing. There are laws that protect participants from 
discrimination based on their genetic information. A Federal law called the Genetic Information 
Nondiscrimination Act (GINA) makes it illegal for health insurance companies, group health plans, 
and most employers to discriminate against individuals based on their genetic information. GINA does 
not protect against genetic discrimination by companies that sell life insurance, disability insurance, or 
long-term care insurance. GINA also does not protect against discrimination based on an already-
diagnosed genetic condition or disease. In some cases, employers could use genetic information to 
decide whether to hire or fire an individual. There is a risk that someone could get access to participant 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
23 
 
genetic information and identify them by name. PUER Research will protect these data in the study and 
will not share or sell these data to outside parties without participants’ written consent.  
 
6.4. 
Quality Control 
 
PUER Research will fulfill the responsibilities identified in their standard operating procedures.  These 
responsibilities include collecting and tracking data forms and instituting quality control measures for 
data entry verification and study compliance.  PUER Research or its designee will be responsible for 
auditing the database and confirming the overall integrity of the data.  PUER Research or its designee 
will ensure that all information pertaining to significant new developments and unanticipated AEs are 
provided to the appropriate regulatory authorities, the Investigators, and to the WCG IRB. 
 
Monitoring of the study will be conducted at regular intervals in order to monitor study conduct and 
maintain GCP by periodic review of the data in the EDC. These inspections are conducted in order to 
verify adherence to the protocol and the completeness and accuracy of the data being entered into the 
CRF. 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
24 
 
 
7. Statistical Considerations 
 
By design, the study will not employ formal hypothesis testing. Study endpoints will be described using 
descriptive statistics, using means and standard deviations, medians with inter-quartile ranges, and 
other descriptive statistical methods, as appropriate. 
 
Data from participants who discontinue the study prior to completion will only be considered through 
the date of withdrawal and will not be followed in the remaining longitudinal study endpoints. 
 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
25 
 
 
8. Ethical, Regulatory, and Administrative Considerations 
 
8.1. 
Informed Consent 
 
The principles of informed consent are described in the Code of Federal Regulations 21 CFR, part 50 
and 45 CFR, part 46.  Once the Investigator has determined the patient’s eligibility for the study, the 
background of the proposed study and the benefits and risks of the procedures and study must be 
explained to the patient.  The patient must be able to comprehend the informed consent form and must 
sign it prior to performing any study specific procedures or prior to receiving medication.  The patient 
and the Investigator will receive a copy of the informed consent.  The original signed informed consent 
and Authorization for Use/Disclosure of PHI will be maintained in the patient’s research chart.  Only 
those patients who sign the IRB approved informed consent prior to participation are eligible to be in 
the study.  Failure to provide written informed consent renders the patient ineligible for the study. 
 
8.2. 
Confidentiality 
 
All information and data collected and/or sent to PUER Research concerning patients or their 
participation in this study will be considered confidential.  Only authorized PUER Research personnel 
will have access to these confidential files.  Authorized personnel have the right to inspect and copy all 
records pertinent to this trial.  All data used in the analysis and reporting of this evaluation will be 
without identifiable reference to any patient. 
 
8.3. 
Institutional Review 
 
The Principal Investigator will obtain approval for the study from the WCG IRB. All changes to the 
protocol must be reviewed and approved prior to implementation.  The Principal Investigator will be 
responsible for obtaining annual renewal through the duration of the study, or more frequently if 
required by the WCG IRB.  PUER Research will maintain all regulatory documents. 
 
8.4. 
Protocol Interpretation and Compliance 
 
The procedures defined in the protocol will be carefully reviewed by the Investigator and research staff 
prior to the time of study initiation to ensure appropriate interpretation and implementation.  No 
deviations from the protocol will be acceptable.  Any changes to the protocol in the form of an 
amendment must be submitted to PUER Research. 
 
8.5. 
Completion of Case Report Forms (CRFs) 
 
The Principal Investigator or his designee will be responsible for completing, in a timely manner, a 
CRF for each patient who is registered to participate in this study.  The Principal Investigator will sign 
and date the indicated places on the CRF.  This signature will indicate that a thorough inspection of the 
data therein has been made and will thereby certify the contents of the form. 
 
 
 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
26 
 
8.6. 
Maintenance of Study Documentation 
 
It is the responsibility of the Principal Investigator and PUER Research staff to maintain a 
comprehensive and centralized filing system of all study-related documentation, which is suitable for 
inspection at any time by the FDA.  Elements should include: 
• 
Participant files – containing the completed CRFs, supporting source documentation, 
and the Informed Consent. 
• 
Regulatory Files – containing the protocol with all amendments and accountability 
records. 
 
8.7. 
Publication Policy 
 
The results of this study will help our understanding of human health, disease, and healthy longevity 
via individualized and longitudinal use of clinical evaluation, biochemical and “omics” assessments, 
non-invasive imaging assessments. PUER Research may publish the results of this research. However, 
names and other identifying information will be kept confidential. This study and its results will not be 
posted to www.clinicaltrials.gov or submitted to the FDA.   
 
8.8. 
Financing and Insurance 
 
Clinical trial insurance will be provided for by PUER Research although it is not expected a 
participant will suffer any study-related injury. Insurance will be obtained and maintained for this 
type of observational study. 
 
8.9. 
Final Study Report 
 
Upon completion of the study, the Principal Investigator is required to submit a final study summary 
report for the patients enrolled in the study. 
 
8.10. 
Record Retention 
 
All records, which are part of this study, will be retained for a period of two years following 
discontinuation/termination of the study. 
 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
27 
 
 
 
9. References 
 
Amendola, L. M., Dorschner, M. O., Robertson, P. D., Salama, J., Hart, R., Shirts, B. H., . . . Jarvik, G. P. 
(2015). Actionable exomic incidental findings in 6503 participants: challenges of variant 
classification. Genome Research, 25(3), 305-315. 
American College of Radiology. (2021). ACR Manual on contrast media. ACR. 
Anthimopoulos, M., Christodoulidis, S., Ebner, L., Christe, A., & Mougiakakou, S. (2016). Lung Pattern 
Classification for Interstitial Lung Diseases Using a Deep Convolutional Neural Network. IEEE 
Transactions on Medical Imaging, 1207-1216. 
Arnoldi, E., Arnoldi, E., Gebregziabher, M., Schoepf, U. J., Goldenberg, R., Ramos-Duran, L., . . . Thilo, 
C. (2010). Automated computer-aided stenosis detection at coronary CT angiography: initial 
experience. European Radiology, 20(5), 1160-1167. 
Assarsson, E., Lundberg, M., Holmquist, G., Björkesten, J., Bucht Thorsen, S., & al., e. (2014). 
Homogenous 96-Plex PEA Immunoassay Exhibiting High Sensitivity, Specificity, and Excellent 
Scalability. PLoS One, 9(4), e95192. 
Bennuru, S., Lustigman, S., Abraham, D., & Nutman, T. B. (2017). Metabolite profiling of infection-
associated metabolic markers of onchocerciasis. Molecular and Biochemical Parasitology, 215, 
58-69. 
Bromberg, Y. (2013). Building a genome analysis pipeline to predict disease risk and prevent disease. 
Journal of Molecular Biology, 425(21), 3993-4005. 
Chong, C.A.K.Y., Tomlinson, G., Chodirker, L., Figdor, N., Uster, M., Naglie, G., Krahn, M.D. (2006). An 
unadjusted NNT was a moderately good predictor of health benefit. Journal of Clinical 
Epidemiology, 59(3):224-33. 
Crick, F. (1970). Central dogma of molecular biology. Nature, 227(5258). 
David C.E.B., Lucas A.M.B., Cunha P.L.O., Viana Y.I.P., Yoshinaga M.Y., Miyamoto S., …  Facundo H.T. (2022) 
Calorie restriction changes lipidomic profiles and maintains mitochondrial function and redox balance during 
isoproterenol-induced cardiac hypertrophy. Journal of Physiology and Biochemistry. Epub ahead of print. 
 
"Design and baseline results of the Scandinavian Simvastatin Survival Study of patients with stable angina 
and/or previous myocardial infarction". (1993). The American Journal of Cardiology. 71(5):393–
400. 
Dorschner, M. O., Amendola, L. M., Turner, E. H., Robertson, P. D., Shirts, B. H., Gallego, C. J., . . . Jarvik, 
G. P. (2013). Actionable, Pathogenic Incidental Findings in 1,000 Participants’ Exomes. American 
Journal of Human Genetics, 93(4), 631-640. 
Fest, J., Vijfhuizen, L. S., Goeman, J. J., Veth, O., Joensuu, A., Perola, M., . . . Hoen, P. A. (2019). Search 
for early pancreatic cancer blood biomarkers in five European prospective population biobanks 
using metabolomics. bioRxiv, 543686. 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
28 
 
Ganz, P., Heidecker, B., Hveem, K., Jonasson, C., Kato, S., Segal, M. R., . . . Williams, S. A. (2016). 
Development and Validation of a Protein-Based Risk Score for Cardiovascular Outcomes Among 
Patients With Stable Coronary Heart Disease. JAMA, 315(23), 2532–41. 
Glasziou, P., & Haynes, B. (2005). The paths from research to improved health outcomes. BMJ Evidence-
Based Medicine(10), 4-7. 
Gold, L. e. (2010). Aptamer-based multiplexed proteomic technology for biomarker discovery. PPLoS One, 
5(12), e15004. 
Goldstein, T. C. (2013). Tools for Extracting Actionable Medical Knowledge from Genomic Big Data. 
Retrieved 9 27, 2021, from https://escholarship.org/uc/item/53k1x0f5 
Guttmacher, A. E., & Collins, F. S. (2003). Welcome to the Genomic Era. The New England Journal of 
Medicine, 349(10), 996-998. 
Han, X., Aslanian, A., Aslanian, A., & Yates, J. R. (2008). Mass Spectrometry for Proteomics. Current 
Opinion in Chemical Biology, 12(5), 483-490. 
Hayden, E. C. (2014). Technology: The $1,000 genome. Nature, 507(7492), 294-295. 
Lemaitre, G., Martí, R., Freixenet, J., Vilanova, J. C., Walker, P., & Meriaudeau, F. (2015). Computer-
Aided Detection and diagnosis for prostate cancer based on mono and multi-parametric MRI. 
Computers in Biology and Medicine, 60, 8-31. 
Li, J., Huang, A., Yao, J., Liu, J., Uitert, R. L., Petrick, N., & Summers, R. M. (2008). Optimizing computer-
aided colonic polyp detection for CT colonography by evolving the Pareto fronta. Medical Physics, 
36(1), 201-212. 
Netto, G., & Kaul, K. e. (2019). Genomic Applications in Pathology. Springer International Publishing. 
Onal, S., Lai-Yuen, S. K., Bao, P., Weitzenfeld, A., & Hart, S. (2014). Fully automated localization of 
multiple 
pelvic 
bone 
structures 
on 
MRI. 
Retrieved 
9 
27, 
2021, 
from 
https://ncbi.nlm.nih.gov/pubmed/25570709 
“Randomised trial of cholesterol lowering in 4444 patients with coronary heart disease: the Scandinavian 
Simvastatin Survival Study (4S”). Lancet. 1994 Nov 19;344(8934):1383-9. PMID: 7968073. 
Rosenberg, S., Elashoff, M., Beineke, P., Daniels, S., Wingrove, J., Tingley, W., . . . Topol, E. (2010). 
Multicenter validation of the diagnostic accuracy of a blood-based gene expression test for 
assessing obstructive coronary artery disease in nondiabetic patients. Annals of internal medicine, 
153(7), 425-434. 
Scaria, V. (2014). Personal genomes to precision medicine. Molecular Cytogenetics, 7(1), 1-1. 
Schork, N. (2015). Personalized medicine: Time for one-person trials. Nature(520), 609-611. 
Uppot, R. N., Boas, G., & Flores, E. (2018). Current Developments in Artificial Intelligence and Diagnostic 
Imaging. 
Retrieved 
9 
27, 
2021, 
from 
https://www.massgeneral.org/assets/MGH/pdf/imaging/radiology-rounds/radiology-rounds-may-
2018.pdf 

PUER Research, LLC 
 
Version 2.0 
PLI001 
 
06Oct2022 
 
CONFIDENTIAL 
 
29 
 
Yang, Q., Li, L., Zhang, J., Shao, G., Zhang, C., Zheng, B., & Zheng, B. (2014). Computer-Aided Diagnosis 
of Breast DCE-MRI Images Using Bilateral Asymmetry of Contrast Enhancement Between Two 
Breasts. Journal of Digital Imaging, 27(1), 152-160. 
Yet, I., Menni, C., Shin, S. Y., Mangino, M., Soranzo, N., Adamski, J., . . . Bell, J. T. (2016). Genetic 
Influences on Metabolite Levels: A Comparison across Metabolomic Platforms. PLoS One, 11(4), 
e0153672. 
Zhu, B., Liu, J. Z., Cauley, S. F., Rosen, B. R., & Rosen, M. S. (2018). Image reconstruction by domain-
transform manifold learning. Nature, 555(7697), 487-492.