Introducing A New Clinical Tool To Assist With

Coronary Artery Disease Risk Assessment

www.prevu.com www.miraculins.com

by

SKIN CHOLESTEROL

ELEVATED

The World’s First and Only Non-Invasive Skin Cholesterol Test

WWW.PREVU.COM

PREVU® HANDHELDSPECTROPHOTOMETER AND

POC TEST REAGENT KIT

COMPLETELY NON-INVASIVE Utilizes safe reagents

CONDUCTED ON THE PALM Patient observes test on the hand

ABSOLUTELY PAINLESS No discomfort whatsoever

NO NEEDLE PRICK OR BLOOD DRAW The skin is not punctured

NO BIOHAZARDOUS MATERIALS Protection not required for test operators

REQUIRES NO OVERNIGHT FASTING Test accurately at any time

TAKES LESS THAN 5 MINUTES Convenient, rapid and streamlined

IMMEDIATE TEST RESULTS No waiting for lab processing

PreVu® Point of Care Test Benefits

www.miraculins.com

by

NO BIOHAZARDOUS MATERIALS Protection not required for test operators

REQUIRES NO OVERNIGHT FASTING Test accurately at any time

TAKES LESS THAN 5 MINUTES Convenient, rapid and streamlined

IMMEDIATE TEST RESULTS No waiting for lab processing

Dear Colleagues:

As you are aware, coronary artery disease (CAD) is one of the leading causes of death in Canada and a highly difficult condition to detect. Evaluation for CAD should ideally begin in a clinical environment where adults would undergo formal risk assessment, including blood cholesterol testing and global risk assessment, under the direction of a healthcare provider. However, many remain unassessed or avoid blood testing entirely. This can result in a potentially dangerous lack of awareness of the significant risk a person may be living with for a coronary event.

Consequently, the earlier identification of individuals at risk in the general population, who could benefit from more intensive primary prevention measures to help guard against a first event, is of primary concern.

The PreVu® Non-Invasive Skin Cholesterol Point of Care (POC) Test is a Canadian developed, non-invasive risk assessment technology designed to measure epidermal sterol, or skin cholesterol – an emerging biomarker in the risk assessment of CAD.

The skin contains 11%, by weight, of total body cholesterol1 and is derived from epithelial steroidogenesis and specific uptake from circulation. Elevated skin cholesterol levels have been shown in clinical studies to be strongly associated with an increased risk of significant CAD as measured by treadmill stress testing2 and coronary angiography3, as well as measured by testing for coronary calcium4, carotid artery thickening5 and carotid artery plaque6. I was involved in early research in this area published in the American Journal of Cardiology in 2002 where skin cholesterol showed significant and positive correlations with systolic blood pressure, LDL-C and the Framingham CAD risk calculation model. We also found that skin cholesterol correlated significantly with inflammatory markers, particularly ICAM-1*.

The following presentation briefly outlines the:

• science and technology behind the PreVu POC Test• test utility and applications• testing procedure step-by-step• mechanism of action• interpretation of results and correlated CAD risk ratios• summary of key clinical references

The PreVu® POC Test does not diagnose the presence or absence of CAD, but has been shown to be effective in helping to identify individuals from the general population who may be at higher, hidden risk for CAD by possessing elevated levels of skin cholesterol.

Sincerely,

Dr. G.B. John Mancini, MD, FRCPC, FACP, FACC (Cardiologist) Professor of MedicineUniversity of British Columbia,Department of Medicine, Division of Cardiology

Director, Cardiovascular Imaging Research Core Laboratory (CIRCL)University of British Columbia, Department of Medicine

Chair, PreVu Medical Advisory BoardNote: References 1-6 appear at end of the booklet

*Mancini J, Chan S, Frochlich J, et al. Association of skin cholesterol content, measured by a non-invasive method, with markers of inflammation and Framingham risk prediction. Am J Cardiol 2002;89:1313-16

FOReWORD

www.miraculins.com

An adhesive foam pad with three wells is placed on the palm of the hand by a test operator. Drops of safe reagent are then placed into the wells in sequence, which first bind with the cholesterol in the skin and then subsequently change color in proportion to the amount of bound cholesterol present. This color change is then read by the PreVu Handheld Spectrophotometer, which is a sophisticated optical reading instrument that also guides the test operator step-by-step through the procedure. The spectrophotometer assigns a numerical value to the color hue and that number places the patient into a range, and accordingly a PreVu risk category (Normal, Borderline, Elevated) based on the clinical data for the biomarker. A PreVu score of 110 or higher (Elevated) may mean an increased risk of coronary artery disease.

The PreVu® POC Test can quickly identify individuals with hidden, higher levels of risk of CAD by testing their skin cholesterol levels non-invasively and without blood draw, discomfort or fasting. It is ideal for delivery through neighborhood healthcare settings including pharmacies, medical clinics, as well as at cardiovascular screening clinics, where in 5 minutes an individual can receive a ‘preview’ of their possible risk of CAD. Then, if warranted, at risk individuals can start down the path towards formal global risk evaluation, primary prevention and ongoing monitoring of all their relevant risks under their healthcare provider’s direction and guidance. Skin Cholesterol testing is not the same as blood serum cholesterol testing, there is no correlation between the results, and a skin cholesterol test is not a substitute for a blood serum cholesterol test.

HOW THe PReVu® POC TeST WORkS

PReVu® POC TeST STeP-By-STePStep 1:Clean Palm; Apply Foam Pad

Step 3:Add Positive Control Reagent

Step 2:Add Detector Reagent

Step 6:Read Degree of Color Hue

Step 5:Add Indicator Reagent

Step 4:Blot Excess Reagents

PReVu® POC TeST uTIlITy AND APPlICATIONS

H3C CH3

CH3H3C

H2N NH2

PReVu® POC TeST MeCHANISM OF ACTION

This image shows the Mechanism of Action by which the Detector reagent - copolymer of Digitonin conjugated with Horseradish Peroxidaise (HRP) - binds with the cholesterol of the skin and then interacts with the Indicator reagent Tetramethylbenzidine (TMB) to produce a color change, which is then read and quantified by the PreVu Handheld Spectrophotometer.

Specificity based on the affinity of Digitonin for the 3b-hydroxy ring structure of cholesterol. The reporter molecule is HRP. The oxidized product of TMB has a deep blue color. The intensity of the blue color - the hue of the TMB solution - is measured directly off the hand by the PreVu Handheld Spectrophotometer.

3,3’,5,5’- TeTRAMeTHylBeNzIDINe (TMB)

INTeRPReTATION OF PReVu® POC TeST ReSulTS

PreVu® Score below 100

Skin Cholesterol risk marker is within normal range

Normal Skin Cholesterol Level

Elevated Skin Cholesterol LevelPreVu® Score 110 or higherElevation correlates with an increased

risk of Coronary Artery Disease

You do not have abnormal skin cholesterol. Further concerns about cardiovascular health related to risk factors such as a family history of cardiovascular disease or high blood cholesterol, being personally overweight, having diabetes, being a smoker, living a sedentary lifestyle or having high blood pressure, should be discussed by the patient with a healthcare provider.

Discuss this test result with your healthcare provider.Formal risk assessment is warranted, including blood cholesterol testing and global risk assessment, under the direction of a healthcare provider.

Borderline Skin Cholesterol Level PreVu® Score between 100 – 109

Discuss this test result with your healthcare provider. Due to test variability of approximately 10%, formal risk assessment, including blood cholesterol testing and global risk assessment under the direction of a healthcare provider, should also be considered for patients with a borderline skin cholesterol reading.

110

100

Individuals with elevated skin cholesterol levels as measured by the PreVu® POC Test, have been shown to have a much greater risk of scoring an abnormal result in the clinical cardiovascular tests listed below:

eleVATeD SkIN CHOleSTeROl AND RISk OF CORONARy ARTeRy DISeASe

RISK OF ABNORMAL CARDIOVASCULAR TESTS WITH ELEVATED SKIN CHOLESTEROL AS MEASURED BY THE PREVU® POC TEST

TEST MEASURES

Functional cardiovascular disease

Coronary artery atherosclerosis

Calci�ed coronary plaque

Carotid artery atherosclerosis

Carotid artery atherosclerosis

IF TEST RESULT ABNORMAL

Possible coronary artery disease

Blockage of coronary arteries

Higher risk of future cardiac events

3.0 times greater risk of abnormal Coronary Calcium4

Increased risk of heart attack and stroke

Increased risk of heart attack and stroke

RISK OF ABNORMAL CLINICAL CARDIOVASCULAR TEST WITH ELEVATED SKIN CHOLESTEROL

2.6 times greater risk of abnormal Treadmill Stress Test2

1.4 times greater risk of abnormal Coronary Angiography3

2.0 times greater risk of abnormal Carotid Ultrasound5 (IMT)

3.0 times greater risk of abnormal Carotid Ultrasound6 (Plaque)

Note: The PreVu® POC test score does not correlate with blood serum cholesterol testing and is not a replacement for blood serum cholesterol testing.

Note: Clinical study references appear at the end of the booklet.

INTeRPReTATION OF PReVu® POC TeST ReSulTS

eleVATeD SkIN CHOleSTeROl AND RISk OF CORONARy ARTeRy DISeASe

The specific clinical references made in this booklet appear below in their order of presentation:

(1) Sabine, JR (1977) Cholesterol, New York: Marcel Dekkar. (2) Zawydiwski R, Sprecher DL, Evelegh MJ, et al. A novel test for the measurement of Skin Cholesterol. Clinical Chemistry 2001;47:1302-4. (3) Sprecher DL, Goodman SG, Kannampuzha P, et al. Skin Tissue Cholesterol (Skin Tc) is related to angiographically-defined cardiovascular disease. Atherosclerosis 2003;171:255-258. (4) Vaidya D, Ding J, Hill JG, et al. Skin Tissue Cholesterol assay correlates with presence of coronary calcium. Atherosclerosis 2005;181:167-73. (5) Tzou WS, Mays ME, Korcarz CE, et al. Skin Cholesterol Content Identifies Increased Carotid Intima-Media Thickness in Asymptomatic Adults. Am Heart J 2005;150:1135-39. (6) Stein JH, Tzou WS, DeCara JM, et al. Usefulness of increased Skin Cholesterol to identify individuals at increased cardiovascular risk (from the Predictor of Advanced Subclinical Atherosclerosis Study). Am J Cardiol 2008;101:986-991.

Since 1999, there have been multiple studies, publications, scientific presentations, and abstracts and posters related to skin cholesterol and the PreVu® technology developed to measure it, that have been conducted, submitted, prepared and delivered by respected researchers from leading North American institutions in cardiovascular research including:

• University of Wisconsin, School of Medicine

• The Cleveland Clinic Foundation

• The Johns Hopkins School of Medicine

• University of Pennsylvania, Department of Experimental Therapeutics

Much of this clinical data has been published in prestigious peer reviewed journals that include:

• The American Journal of Cardiology

• The American Heart Journal

• Atherosclerosis

• Clinical Chemistry

Corresponding presentations have been made at some of the most important conferences and meetings in the clinical world including:

• Annual American Association of Clinical Chemistry Meeting

• American Heart Association Scientific Sessions

• Canadian Cardiovascular Congress

• Annual Scientific Session of the American College of Cardiology

• The Endocrine Society Annual Meeting

• American Heart Association Conference on Arteriosclerosis, Thrombosis and Vascular Biology

For further clinical evidence please visit www.prevu.com.

ClINICAl ReFeReNCeS

Printed in Canada.

For further information on the science of skin cholesterol, the PreVu technology

and the clinical applications of both in risk assessment of coronary artery disease,

please visit www.prevu.com

www.miraculins.com

by

Copyright © 2013 by Miraculins Inc. All Rights Reserved. Unauthorized distribution, transmission or republication is strictly prohibited. PreVu® is a Registered Trademark of Miraculins Inc. All Rights Reserved. 2013.

For further information on distributing the PreVu® POC Test technology in your community, please contact:

Paul J. MoreauVice President, Sales and MarketingMiraculins Inc.T. 204.478.5603 • F. 204.453.1546pmoreau@miraculins.com