PIVUS

The PIVUS study was initiated in 2001 as a research collaboration between the Department of Medical Sciences, Uppsala University Hospital and AstraZeneca R&D Mölndal with the primary aim to evaluate the usefulness of differernt measurements of endothelial function and other techniques to evaluate vascular function.

Several secondary aims have been added to this prospective cohort study and several academic groups have been engaged in the evaluation of this cohort from different aspects.

List divided according to fields of interest.

• Agneta Siegbahn, coagulation, Uppsala University Hospital
• Anders Larsson, clinical chemistry, Uppsala University Hospital
• Tord Neassen, vascular function and imaging, Uppsala University Hospital
• Kenny Rodriguez, vascular function and imaging, Uppsala University Hospital
• Johan Sundström, cardiovascular epidemiology, Uppsala University Hospital
• Bengt Vessby, nutrition and metabolism, Uppsala University Hospital
• Karl Mikaelsson, osteroporosis, Uppsala University Hospital
• Håkan Melhus, osteoporosis and genetics, Uppsala University Hospital
• Andreas Kindmark, osteoporosis, Uppsala University Hospital
• Johan Ärnlöv, myocardial function, Uppsala University Hospital
• Ann-Christine Syvänen, genetic analysis, Uppsala University Hospital
• Samar Basu, inflammation and oxidative stress, Uppsala University Hospital
• Lisa Kurland, genetics and hypertension, Uppsala University Hospital
• Anders Karlsson, metabolism and obesity, Uppsala University Hospital
• Anders Holmlund, Peridontitis, Gävle Hospital
• Ann-Kari Lefvert, immunology, CMM, Karoliska Hospital
• Johan Frostegård, immunology, Karoliska Hospital
• Stefan Söderberg, obesity related hormones, Umeå university Hospital
• Bo Sandhagen, rheology, Uppsala University Hospital
• Jessika Andersson, plaque characterization, Uppsala University Hospital
• Anna Stenborg, endothelial function and stroke, Uppsala University Hospital
• Johannes Hulthe, inflammation and atherosclerosis, Sahlgrenska Hospital
• Håkan Ahlström, head of MRI unit, Uppsala University Hospital
• Thomas Bjerner, myocardial MRI, Uppsala University Hospital
• Lars Johansson, MRI, Uppsala University Hospital
• Thomas Hansen, PhD student MRI of the vasculature, Uppsala University Hospital
• Thomas Kahan, stress hormones and myocardium, Danderyds Hospital
• Mikel Zilmer, oxidation/antioxidation, Tartuu Hospital, Estonia
• Bertil Lindahl, risk markers for myocardial ischemia, Uppsala University Hospital
• Lars Wallentin, risk markers for myocardial ischemia, Uppsala University Hospital
• Charlotte Ebeling Barbier, Myocardial MRI, Uppsala University
• Erik Ingelsson, Adidokines, Uppsala University

Age 70 measurements were the following:

1. Endothelial function. Three different tests of endothelium-dependent vasodilation were performed:

• The invasive forearm technique using intra-brachial artery infusion of acetylcholine and SNP to evaluate the increase in forearm blood flow by venous occlusion plethysmography.
• Flow-mediated vasodilation (FMD) of the brachial artery using ultrasound.
• Change in the reflected pulse waves obtained by aplanation tonometry (Sphygmocor) following sc given terbutaline.

2. Arterial compliance. Three different tests of arterial compliance were performed:

• Distensibility of the carotid artery evaluated by ultrasound.
• The stroke volume/pulse pressure ratio (SV/PP) evaluated by echocardiography.
• The Augmentation index (AIx) evaluated by pulse wave analysis.

3. Echocardiography. A standard echocardiography with Doppler examination was performed to evaluate LV geometry, systolic and diastolic function. Variables collected: LVEDD, IVS, PW, LV mass, SI, CI, EF, IVRT, E/A ratio, ejection time, myocardial performance index (MPI).

4. Blood pressure was measured by sphygmomanometer and intra-arterially. Blood pressure was also measured in the ankles for calculation of the ABI in the last 500 of the cohort.

5. Anthropometry. Height, weight, waist, hip and sagital diameter.

6. ECGwas taken as a conventional 12 lead ECG and subjected to Minnesota coding. A 6-lead precordial ECG was also recorded continuously in digital form for 5 min and subjects to analysis in the Eclysis software. By this analysis we have data on variability of the various time intervals and amplitudes detected at the ECG both spatially and temporarily. So far, temporal QT-variability and spatial QT dispersion has been used.

7. Baroreceptor sensitivity and heart rate and blood pressure variability. By combining the information from continuously recorded ECG and invasive blood pressure, baroreceptor sensitivity was calculated by both the sequence method and the alpha-index method. By applying spectral analysis variability in the frequency domain has been calculated for heart rate and blood pressure.

8. Ultrasound of the carotid arteries was performed for determination of IMT and plaque occurrence. An analysis is undergoing to characterize the plaque according to grey scale analyis.

In 100 subjects intima and media were investigated separately by a 25 MHz probe.

9. DXA was performed in 900 subjects 1-2 years following the main examination. From this bone mineral density, lean body mass and body fat could be calculated from the whole body as well as from different parts of the body. Data are under preparation.

10. Lung function tests were performed in 900 subjects 1-2 years following the main examination. Vital capacity, FEV1 and PEF were recorded.

11. Cardiac MRI was performed in 300 subjects 1-2 years following the main examination. From this, information is available regarding late enhancement (myocardial scars), LV mass, EF, AVPD.

12. Whole body MR angiography was performed in 300 subjects 1-2 years following the main examination. In this examination there is information on atherosclerosis in the carotid arteries, the aorta, the renal arteries, the femoral part of the leg as well as the lower leg. A composite score for these vascular segments have been calculated.

13. Abdominal fat MR. A MRI scan was performed at the L4 level for calculations of subcutaneous (SAT) and visceral fat area (VAT) in 300 subjects 1-2 years following the main examination.

14. DNA has been prepared from leukocytes and genotyping for SNPs in candidate genes are soon to begin.

15. Serum /plasma sampling. A number of serum/plasma samples were collected and placed in freezers for later analysis. These laboratory tests have been divided in different categories.

a.Routine haematology: Hemoglobin, hematocrite, leukocyte count and platelet count.

b.Routine blood chemistry: liver function tests, kidney function, electrolytes, iron and transferring, urate.

c.Rheology: Whole blood and plasma viscosity, erythrocyte aggregability and fluidity.

d.Lipids: Serum cholesterol and triglycerides, HDL and LDL-cholesterol, apoB/A1 ratio (apoE and oxLDL under preparation)

e.Inflammation: hsCRP (MCP-1 and a number of cytokines are under preparation)

f.Adipokines: Leptin and adiponectin

g.Oxidation/Antiox: Conjugated diens, lag phase LDL, antibodies LDL, glutathione , oxidized and redox,. total antiox capacity, homocysteine.

h.Coagulation: vWillebrant factor and PAI-1 under preparation

i.Immunology: Cardiolipin antibodies, circulating immune complexes, and T-cell markers.

j.Collagen turnover: MMP-9 and TIMP-1

k.Calcium metabolism: PTH and serum calcium, phosphate and magnesium

l.Glucos heomeostasis: fasting blood glucose and fasting insulin

m.NO synthesis: ADMA, L-arginine under preparation

16. Food intake records for 7 days are available for 850 subjects. There is data on different nutrients, including different fatty acids, as well as different dietary sources.

17. Self-reported history of diseases and medication, as well as a number of other items regarding life-style, such as exercise, smoking, alcohol intake, education, social network, number of teeth etc.

PIVUS measurements at age 75:

1. Endothelial function. Two different tests of endothelium-dependent vasodilation will be performed:

a. Flow-mediated vasodilation (FMD) of the brachial artery using ultrasound.

b. Change in the reflected pulse waves obtained by aplanation tonometry (Sphygmocor) following sc given terbutaline.

2. Arterial compliance. Three different tests of arterial compliance will be performed:

a. Distensibility of the carotid artery evaluated by ultrasound.

b. The stroke volume/pulse pressure ratio (SV/PP) evaluated by echocardiography.

c. The Augmentation index (AIx) evaluated by pulse wave analysis.

3. Echocardiography. A standard echocardiography with Doppler examination will be performed to evaluate LV geometry, systolic and diastolic function.

Variables collected: LVEDD, IVS, PW, LV mass, SI, CI, EF, IVRT, E/A ratio, ejection time, myocardial performance index (MPI).

4. Blood pressure will be measured by sphygmomanometer in the arm. Blood pressure will also be measured in the ankles for calculation of the ABI

5. Anthropometry. Height, weight, waist, hip and sagital

diameter.

6. ECGwill be taken as a conventional 12 lead ECG and subjected to Minnesota coding.

7. Ultrasound of the carotid arteries will performed for determination of IMT and plaque occurrence and grey scale analysis.

In addition, the femoral arteries will be investigated in the same manner and intima and media will be investigated separately in the carotid arteries by a 25 MHz probe.

8. Lung function tests will be performed: Vital capacity, FEV1 and PEF will be recorded.

9. Cardiac MRI will be performed in the 300 subjects measured at age 70.From this, information is available regarding late enhancement (myocardial scars), LV mass, EF, AVPD.

10. Whole body MR angiography will be performed in the 300 subjects measured at age 70.In this examination there is information on atherosclerosis in the carotid arteries, the aorta, the renal arteries, the femoral part of the leg as well as the lower leg. A composite score for these vascular segments have been calculated.

11. Abdominal fat MR will be performed in all subjects. A MRI scan will be performed of the abdomen for calculations of subcutaneous (SAT) and visceral fat volume (VAT).

12. Serum /plasma sampling. A number of serum/plasma samples will be collected and placed in freezers for later analysis.

Routine haematology, lipids and fasting blood glucose will be rapidly analysed for feed-back to the subjects.

13. Self-reported history of diseases and medication, as well as a number of other items regarding life-style, such as exercise, smoking, alcohol intake, education, social network, number of teeth etc.

14. Cognition test. Three different test for cognition will be applied: MMSE, TMT

15. Brain MRI will be conducted in the total cohort for measurements of hippocampus volume, lucunar infarcts, bleedings, leukaryosis and peripheral brain infarcts.

16. Carotid artery MRI will be conducted in the total cohort for measurements of carotid wall volume, plaque composition and contrast enhancement for detection of neovascularisation and inflammation.

17. Ultrasound of abdominal aorta will be performed for screening of aortic aneurysms.