Chronic kidney disease (CKD), is characterized by accelerated development of atherosclerosis and advanced remodelling of vessels and the heart. It is associated with many factors, including inflammation, arterial hypertension, hyperlipidemia, hyperhomocysteinemia, secondary hyperparathyroidism, and oxidative stress. Hypertension is one of the most critical risk factors for cardiovascular complications. It leads to the formation of structural changes in the vascular system: it impairs the activity of the endothelium, causes hypertrophy and remodelling of the vascular wall, reduces the susceptibility of the vessels and accelerates the development of atherosclerosis. This study aimed to identify the processes and their representative markers, the concentration of which in the serum may reflect the cardiovascular system status and can predict the increased mortality in HD patients.
Chronic Kidney Disease has a significant impact on the cardiovascular system. From many different complications of CKD, one to mention is arterial stiffness. This disorder results from many pathologies, including inflammation, arterial hypertension, carbohydrate metabolic disorders, lipid disorders, vascular calcification, chronic inflammation, and oxidative stress. The main goal of this study was to analyze the mechanisms leading to the increased tendency to cardiovascular disturbances in CKD, with particular focus on the parameters of oxidative stress, inflammation and the results of imaging examinations (intima-media thickness (IMT) assessments) and other non-invasive cardiological examinations based on the results using the Portapres device (Finapres Medical Systems (FMS), the Netherlands), the SphygmoCor tonometer (AtCor Medical), the Colin blood pressure monitor (BMP)-7000 (Japan) Pulse Trace 2000 (Micro Medical Ltd., Rochester, Kent, United Kingdom) The Accuson CV 70 system (Siemens) with a 10 megahertz (Mhz) transducer. Besides, studied participants were followed 2 years after enrollment to study for recording cardiovascular-related death.
Study Type
OBSERVATIONAL
Enrollment
252
the complete blood count was analyzed using Sysmex K-4500 Automated Hematology Analyzer (by GMI Inc., USA): * hemoglobin (HGB) \[g/dl\]; * red blood count (RBC) \[10\^12/l\]; * hematocrit (HCT) \[l/l\]; * white blood cells (WBC) \[10\^9/l\]; * platelet count (PLT) \[10\^9/l\]
body mass index (BMI) \[kg/m\^2\] was calculated by dividing a person's weight (post-HD weight in HD group) \[kg\] by the squared their body height \[m\]
Serum concentration of: * advanced glycation ends products (AGE) \[µg/mg protein\]; * 3-nitrotyrosine (3-NT) \[µmol/mg protein\]; * advanced oxidation protein products (AOPP) \[µmol/mg protein\]; * carboxymethyle(lysine) (CML) \[µg/mg protein\] were determined with the enzyme immunoassay methods (ELISA) using Shanghai Sunred Biological Technology Co kits, China.
metalloproteinases in the serum \[ng/ml\]: * metalloproteinase 9 (MMP-9) in the serum was determined by the ELISA method using the Quantikine Human MMP-9 (total) kit, by R\&D Systems, Canada; * tissue inhibitor of metalloproteinase 1 (TIMP-1) in the serum - was determined by the ELISA method using the Quantikine Human TIMP-1 kit, manufactured by R\&D Systems, Canada; * the MMP-9/TIMP-1 ratio was calculated by the quotient of the MMP-9 and the TIMP-1 concentration.
* total cholesterol (T-C) \[mg/dl\] - was assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA; * low-density lipoprotein cholesterol (LDL-C) \[mg/dl\] - was assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA; * high-density lipoprotein cholesterol (HDL-C) \[mg/dl\] - was assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA; * triglycerides (TG) \[mg/dl\] - were assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA; * the concentration of low-density lipoprotein (LDL-C) cholesterol - was determined from Friedewalds' equation (LDL-C \[mg/dl\] = total cholesterol (T-C) \[mg/dl\]- HDL-C \[mg/dl\]- TG\[mg/dl\]/5).
* iron concentration \[mg/dl\] - was analyzed with the Cobas Integra 400 plus biochemical analyzer from Roche Diagnostics, USA; * total iron-binding capacity (TIBC) \[mg/dl\] - was analyzed with the Cobas Integra 400 plus biochemical analyzer from Roche Diagnostics, USA; * the unsaturated iron-binding capacity (UIBC) \[mg/dl\] was determined by an equation in which iron concentration in plasma is subtracted from TIBC \[mg/dl\]; * ferritin \[ng/ml\] concentration was determined with the Modular E-170 biochemical analyzer from Roche Diagnostics, USA.
* high-sensitivity C-reactive protein (hsCRP) \[mg/l\] was measured using DADE Behring, USA and the DADE nephelometer Behring Analyzer II; * neopterin \[nmol/l\] was determined by using the Neopterin ELISA kit, DRG International, Inc., USA; * interleukin 18 (IL-18) \[pg/ml\] concentration was determined by Colorimetric Sandwich ELISA, Quantikine Human IL-18 R\&D Inc., USA.
carotid intima-media thickness (IMT) \[mm\] was measured by The Accuson CV 70 system (Siemens) with a 10 megahertz (Mhz) transducer. Two longitudinal projections were assessed (anterolateral and posterolateral). The distal 1 cm of the common carotid artery just proximal to the bulb was measured by means of a computer analysis system (Medical Imaging Applications, LLC).
For non-invasive cardiological examinations, the Portapres device (Finapres Medical Systems (FMS), the Netherlands), the SphygmoCor tonometer (AtCor Medical), the Colin blood pressure monitor (BMP)-7000 (Japan) were used. Main assessed variables: heart rate (HR) \[beats per minute \[bpm\]\]; ejection duration (ED) \[millisecons\]; peripheral systolic (pSBP) and diastolic blood pressure (pDBP) \[mm Hg\]; peripheral mean arterial pressure (pMAP) \[mm Hg\]; peripheral end-systolic pressure (pESP) \[mm Hg\]; central systolic (cSBP) and diastolic blood pressure (cDBP) \[mm Hg\]; central mean arterial pressure (cMAP) \[mm Hg\]; central augmented pressure (cAP) \[mmHg\]; central mean pressure of diastole (cMPD)\[mm Hg\]; central mean pressure of systole (cMPS) \[mm Hg\]; central end-systolic pressure (cESP) \[mm Hg\].
The following parameters of vessel stiffness were assessed by Pulse Trace 2000 (Micro Medical Ltd., Rochester, Kent, United Kingdom): * reflection index (RI) \[in percentages \[%\]\]; * vascular stiffness index (SI) \[m/s\]; * peripheral pulse pressure (pPP) \[mm Hg\]; * central puls pressure (cPP) \[mm Hg\] * peripheral pulse pressure/central pulse pressure (pPP/cPP ratio).
During a 2-year follow-up from the enrollment to this study, CV-related fatal incidents history has been recorded for each subject separately. The primary endpoint was fatal acute myocardial infarction (AMI) or acute ischemic stroke or any unexpected or sudden death only if autopsy proved CV-related. If there was doubt about the cause of death or there was no contact with the patient during the two years from study enrollment, that patient was excluded and not considered further.
glucose (Glu) \[mg/dl\] was assessed in the serum by a routine technique using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
klotho \[ng/ml\] - was analyzed in the serum by Human KL(Klotho) \[ng/ml\] ELISA Kit, Shanghai Sunred Biological Technology Co kit, China.
FGF-23 \[pg/ml\] - was analyzed in rhe serum using Human FGF-23 ELISA Kit, Sigma-Aldrich, USA.
* total and ionized calcium \[mg/dl\], * phosphate \[mg/dl\], * intact parathormone (iPTH) \[mg/dl\] were assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
activity of: * alanine transaminase (ALT) \[U/l\]; * aspartate transaminase (AST) \[U/l\]; * alkaline phosphatase (ALP) \[U/l\] were assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
* total protein (TP) \[g/dl\]; * albumin (ALB) \[g/dl\] were assessed in the serum by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
* creatinine in the serum \[mg/dl\] - the assay is based on the reaction of creatinine with sodium picrate as described by Jaffe (Jaffes' colorimetric method) - was assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA; * urea \[mg/dl\] in the serum - was assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
myeloperoxidase (MPO) \[ng/ml\] in the serum- was determined by the ELISA method using the Quantikine Human MPO test by R\&D Systems kit, Canada.
soluble receptor for advanced glycation end products (sRAGE) \[µg/mg protein\] in the serum was tested with enzymatic immunoassay (Quantikine ELISA) using R\&D Systems kit, Canada.
* methylglyoxal (MG) \[µg/mg protein\]; * carboxyethyle(lysine) (CEL) \[µg/mg protein\]; * carbamyl protein groups \[µg/mg protein\] were assessed in the serum by competitive enzyme immunoassay (competitive ELISA) using kits from Cell Biolabs Inc, USA.
* potassium (K) \[mmol/l\]; * sodium (Na) \[mmol/l\]; * magnesium (Mg) \[mg/dL\] were assessed in the serum by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
NT-proBNP \[fmol/ml\] - was analyzed in the serum by enzyme immunoassay using the Nt-proBNP kit from Biomedica, Slovakia.
eGFR \[ml/min/1.73m\^2\] - according to the Kidney Disease: Improving Global Outcomes (KDIGO) 2012 recommendations was calculated based on the Modification of Diet in Renal Disease (MDRD) formula: eGFR = 186 x \[creatinine concentration in mg/dl\] - 1.154 x \[age in years\] - 0.203 x \[0.724\] for the female gender.
Poznan University of Medical Sciences
Poznan, Poland
Diagnostic test: basic biochemical parameters: complete blood count - hemoglobin (HGB)
hemoglobin (HGB) \[g/dl\] was analyzed using Sysmex K-4500 Automated Hematology Analyzer (by GMI Inc., USA).
Time frame: 3 years
Diagnostic test: basic biochemical parameters: complete blood count - red blood cell count (RBC)
red blood cell count (RBC) \[10\^12/l\] was analyzed using Sysmex K-4500 Automated Hematology Analyzer (by GMI Inc., USA).
Time frame: 3 years
Diagnostic test: basic biochemical parameters: complete blood count - hematocrit (HCT)
hematocrit (HCT) \[l/l\] was analyzed using Sysmex K-4500 Automated Hematology Analyzer (by GMI Inc., USA).
Time frame: 3 years
Diagnostic test: basic biochemical parameters: complete blood count - white blood cell count (WBC)
white blood cells (WBC) \[10\^9/l\] was analyzed using Sysmex K-4500 Automated Hematology Analyzer (by GMI Inc., USA).
Time frame: 3 years
Diagnostic test: basic biochemical parameters: complete blood count - platelet count (PLT)
platelet count (PLT) \[10\^9/l\] was analyzed using Sysmex K-4500 Automated Hematology Analyzer (by GMI Inc., USA).
Time frame: 3 years
Diagnostic test: glucose (Glu)
glucose (Glu) \[mg/dl\] concentration in the serum was assessed by the routine technique using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: urea
urea \[mg/dl\] concentration in the serum was assessed by the routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: creatinine
creatinine \[mg/dl\] concentration in the serum was assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA (based on Jaffes' colorimetric method - the assay is based on the reaction of creatinine with sodium picrate as described by Jaffe).
Time frame: 3 years
Estimated glomerular filtration rate (eGFR) [ml/min/1.73m^2] calculation
eGFR - according to the Kidney Disease: Improving Global Outcomes (KDIGO) 2012 recommendations was calculated based on the Modification of Diet in Renal Disease (MDRD) formula: eGFR = 186 x \[creatinine concentration in mg/dl\] - 1.154 x \[age in years\] - 0.203 x \[0.724\] for the female gender.
Time frame: 3 years
Body mass index (BMI) [kg/m^2] calculation
Body mass index (BMI) - \[kg/m\^2\] was calculated by dividing a person's weight (post-HD weight in HD group) \[kg\] by the squared their body height \[m\].
Time frame: 3 years
Diagnostic test: parameters of lipids metabolism in the serum - total cholesterol (T-C)
total cholesterol (T-C) \[mg/dl\] concentration in the serum was assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: parameters of lipids metabolism in the serum - high-density lipoprotein cholesterol (HDL-C)
high-density lipoprotein cholesterol (HDL-C) \[mg/dl\] concentration in the serum was assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: parameters of lipids metabolism in the serum low-density lipoprotein cholesterol (LDL-C).
low-density lipoprotein (LDL-C) cholesterol concentration in the serum was determined from Friedewals' equation (LDL-C \[mg/dl\] = total cholesterol (T-C) \[mg/dl\] - HDL-C \[mg/dl\] - TG\[mg/dl\]/5). It was assessed by the routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: parameters of lipids metabolism in the serum - triglycerides (TG)
triglycerides (TG) \[mg/dl\] concentration in the serum was assessed by the routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: liver enzymes activity assessment - aspartate transaminase (AST)
activity of aspartate transaminase (AST) \[U/l\]; was assessed in the serum by the routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: liver enzymes activity assessment - alanine transaminase (ALT)
activity of alanine transaminase (ALT) \[U/l\] was assessed in the serum by the routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: liver enzymes activity assessment - alkaline phosphatase (ALP) [U/l]
activity of alkaline phosphatase (ALP) \[U/l\] was assessed in the serum by the routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: total protein (TP)
total protein (TP) \[g/dl\] concentration in the serum was assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: albumin(ALB)
albumin (ALB) \[g/dl\] concentration in the serum was assessed by the routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: parameters of iron metabolism - iron
iron concentration \[mg/dl\] in the serum - was assessed with the Cobas Integra 400 plus biochemical analyzer from Roche Diagnostics, USA;
Time frame: 3 years
Diagnostic test: parameters of iron metabolism - total iron-binding capacity (TIBC)
total iron-binding capacity (TIBC) \[mg/dl\] - was determined with the Cobas Integra 400 plus biochemical analyzer from Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: parameters of iron metabolism - the unsaturated iron-binding capacity (UIBC)
unsaturated iron-binding capacity (UIBC) \[mg/dl\] was determined by an equation in which iron \[mg/dl\] concentration in plasma is subtracted from TIBC \[mg/dl\].
Time frame: 3 years
Diagnostic test: parameters of iron metabolism - ferritin
ferritin \[ng/ml\] concentration in the serum was determined with the Modular E-170 biochemical analyzer from Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: total and ionized calcium
total and ionized calcium \[mg/dl\] serum concentrations were assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: phosphate
phosphate \[mg/dl\] serum concentration was assessed by the routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: intact parathormone (iPTH)
intact parathormone (iPTH) \[mg/dl\] serum concentration was assessed by the routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: klotho (KL)
klotho (KL) \[ng/ml\] serum concentration was analyzed by Human KL(Klotho) \[ng/ml\] ELISA Kit, Shanghai Sunred Biological Technology Co kit, China.
Time frame: 3 years
Diagnostic test: fibroblast growth factor 23 (FGF-23)
fibroblast growth factor 23 (FGF-23) \[pg/ml\] serum concentration was analyzed using Human FGF-23 ELISA Kit, Sigma-Aldrich, USA.
Time frame: 3 years
Diagnostic test: selected electrolytes assessment in the serum: potassium (K) and sodium (Na)
Electrolytes: potassium (K) \[mmol/l\] and sodium (Na) \[mmol/l\] serum concentrations were assessed by the routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: selected electrolytes assessment in the serum: magnesium
magnesium (Mg) \[mg/dl\] serum concentration was assessed by routine techniques using Cobas Integra 400 plus biochemical analyzer by Roche Diagnostics, USA.
Time frame: 3 years
Diagnostic test: selected parameters of oxidative stress - 3-nitrotyrosine (3-NT)
Serum concentration of 3-nitrotyrosine (3-NT) \[µmol/mg protein\] was determined with the enzyme immunoassay method (ELISA) for 3NT using Shanghai Sunred Biological Technology Co kits, China.
Time frame: 3 years
Diagnostic test: selected parameters of oxidative stress - advanced glycation ends products (AGE)
Serum concentration of advanced glycation ends products (AGE) \[µg/mg protein\] was determined with the enzyme immunoassay method (ELISA) for AGE using Shanghai Sunred Biological Technology Co kits, China.
Time frame: 3 years
Diagnostic test: selected parameters of oxidative stress - carboxymethyle(lysine) (CML)
Serum concentration of carboxymethyle(lysine) (CML) \[µg/mg protein\] was determined with the enzyme immunoassay method (ELISA) for CML using Shanghai Sunred Biological Technology Co kits, China.
Time frame: 3 years
Diagnostic test: selected parameters of oxidative stress - advanced oxidation protein products (AOPP)
Serum concentration of advanced oxidation protein products (AOPP) \[µmol/mg protein\] was determined with the enzyme immunoassay method (ELISA) for AOPP using Shanghai Sunred Biological Technology Co kits, China.
Time frame: 3 years
Diagnostic test: metalloproteinases - metalloproteinase 9 (MMP-9)
metalloproteinase 9 (MMP-9) \[ng/ml\] concentration in the serum was determined by the ELISA method using the Quantikine Human MMP-9 (total) kit, by R\&D Systems, Canada.
Time frame: 3 years
Diagnostic test: metalloproteinases - tissue inhibitor of metalloproteinase 1 (TIMP-1)
tissue inhibitor of metalloproteinase 1 (TIMP-1) \[ng/ml\] concentration in the serum - was determined by the ELISA method using the Quantikine Human TIMP-1 kit, manufactured by R\&D Systems, Canada.
Time frame: 3 years
The MMP-9/TIMP-1 ratio assessment
the MMP-9/TIMP-1 ratio was calculated by the quotient of the MMP-9 \[ng/ml\] and the TIMP-1 \[ng/ml\] concentration.
Time frame: 3 years
Diagnostic test: selected inflammatory markers - high-sensivity C-reactive protein (hsCRP)
high-sensitivity C-reactive protein (hsCRP) \[mg/l\] concentration in the serum was measured using DADE Behring, USA, and the DADE nephelometer Behring Analyzer II.
Time frame: 3 years
Diagnostic test: selected inflammatory markers - neopterin
neopterin \[nmol/l\] serum concentration was determined by using the Neopterin ELISA kit, DRG International, Inc., USA.
Time frame: 3 years
Diagnostic Test: selected inflammatory markers - interleukin 18 (IL-18)
interleukin 18 (IL-18) \[pg/ml\] concentration in the serum was determined by Colorimetric Sandwich ELISA, Quantikine Human IL-18 R\&D Inc., USA.
Time frame: 3 years
Diagnostic test: selected parameters of oxidative stress - myeloperoxidase (MPO)
myeloperoxidase (MPO) \[ng/ml\] in the serum - was determined by the ELISA method using the Quantikine Human MPO test by R\&D Systems kit, Canada.
Time frame: 3 years
Diagnostic test: selected parameters of oxidative stress - methylglyoxal (MG)
methylglyoxal (MG) \[µg/mg protein\] concentration in the serum was assessed by competitive enzyme immunoassay (competitive ELISA) using MG kits from Cell Biolabs Inc, USA.
Time frame: 3 years
Diagnostic test: selected parameters of oxidative stress - carboxyethyle(lysine) (CEL) [µg/mg protein]
carboxyethyle(lysine) (CEL) \[µg/mg protein\] concentration in the serum was assessed by competitive enzyme immunoassay (competitive ELISA) using CEL kits from Cell Biolabs Inc, USA.
Time frame: 3 years
Diagnostic test: selected parameters of oxidative stress - carbamyl protein groups [µg/mg protein]
carbamyl protein groups \[µg/mg protein\] concentration in the serum were assessed by competitive enzyme immunoassay (competitive ELISA) using carbamyl protein groups kits from Cell Biolabs Inc, USA.
Time frame: 3 years
Diagnostic test: selected parameters of oxidative stress - soluble receptor for advanced glycation end products (sRAGE)
soluble receptor for advanced glycation end products (sRAGE) \[µg/mg protein\] concentration in the serum was tested with enzymatic immunoassay (Quantikine ELISA) using R\&D Systems sRAGE kit, Canada.
Time frame: 3 years
Non-invasive cardiological examinations (1) with the use of Portapres device (Finapres Medical Systems (FMS), the Netherlands), the SphygmoCor tonometer (AtCor Medical), the Colin blood pressure monitor (BMP)-7000 (Japan) - blood pressures
Blood pressure was measured using the Colin BPM 7000 on both arms (participants were seated). Next, a piezoelectric tonometer Colin BPM was placed over the radial artery for the acquisition of the radial arterial pressure waveform in a supine position. This signal was sent to the SphygmoCor and after averaging various parameters have been assessed and recorded: * peripheral systolic blood pressure (sSBP) \[mmHg\]; * peripheral diastolic blood pressure (pDBP)\[mm Hg\]; * peripheral mean arterial pressure (pMAP) \[mm Hg\]; * peripheral end-systolic pressure (pESP) \[mm Hg\]; * central systolic blood pressure (cSBP) \[mm Hg\]; * central diastolic blood pressure (cDBP) \[mm Hg\]; * central mean arterial pressure (cMAP) \[mm Hg\]; * entral augmented pressure (cAP) \[mm Hg\]; * central mean pressure of diastole (cMPD) \[mm Hg\]; * central mean pressure of systole (cMPS)\[mm Hg\]; * central end-systolic pressure (cESP)\[mm Hg\]
Time frame: 3 years
Non-invasive cardiological examinations (2) with the use of Portapres device (Finapres Medical Systems (FMS), the Netherlands), the SphygmoCor tonometer (AtCor Medical), the Colin blood pressure monitor (BMP)-7000 (Japan) - heart rate (HR)
Blood pressure was measured using the Colin BPM 7000 on both arms (participants were seated). Next, a piezoelectric tonometer Colin BPM was placed over the radial artery for the acquisition of the radial arterial pressure waveform in a supine position. This signal was sent to the SphygmoCor and after averaging various parameters have been assessed and recorded: \- heart rate (HR) in beats per minute \[bpm\]
Time frame: 3 years
Non-invasive cardiological examinations (3) with the use of Portapres device (Finapres Medical Systems (FMS), the Netherlands), the SphygmoCor tonometer (AtCor Medical), the Colin blood pressure monitor (BMP)-7000 (Japan) - ejection duration (ED)
Blood pressure was measured using the Colin BPM 7000 on both arms (participants were seated). Next, a piezoelectric tonometer Colin BPM was placed over the radial artery for the acquisition of the radial arterial pressure waveform in a supine position. This signal was sent to the SphygmoCor and after averaging various parameters have been assessed and recorded: \- ejection duration (ED) in milliseconds \[msec\]
Time frame: 3 years
Device: carotid intima-media thickness (IMT)
Carotid intima-media thickness (IMT) \[mm\] was measured by The Accuson CV 70 system (Siemens) with a 10 megahertz (Mhz) transducer. Two longitudinal projections were assessed (anterolateral and posterolateral). The distal 1 cm of the common carotid artery just proximal to the bulb was measured by means of a computer analysis system (Medical Imaging Applications, LLC).
Time frame: 3 years
Device: vessel stiffness assessments - reflection index (RI)
The following parameter of vessel stiffness was assessed by Pulse Trace 2000 (Micro Medical Ltd., Rochester, Kent, United Kingdom): \- reflection index (RI) in percentages \[%\].
Time frame: 3 years
Device: vessel stiffness assessments - vascular stiffness index (SI)
The following parameter of vessel stiffness was assessed by Pulse Trace 2000 (Micro Medical Ltd., Rochester, Kent, United Kingdom): -vascular stiffness index (SI) \[m/s\].
Time frame: 3 years
Device: vessel stiffness assessments - peripheral (pPP) and central pulse pressure (cPP) [mm Hg]
The following parameters of vessel stiffness were assessed by Pulse Trace 2000 (Micro Medical Ltd., Rochester, Kent, United Kingdom): * peripheral pulse pressure (pPP) \[mm Hg\]; * central pulse pressure (cPP) \[mm Hg\]
Time frame: 3 years
Device: vessel stiffness assessments - peripheral pulse pressure/central pulse pressure (pPP/cPP) ratio
Peripheral pulse pressure/central pulse pressure (pPP/cPP) ratio was assessed by dividing peripheral pulse pressure (pPP) \[mm Hg\] by central pulse pressure (cPP) \[mm Hg\].
Time frame: 3 years
Cardiovascular (CV)-related death recording during 2-year follow-up
During a 2-year follow-up from the enrollment to this study, CV-related fatal incidents history has been recorded for each subject separately. The primary endpoint was fatal acute myocardial infarction (AMI) or acute ischemic stroke or any unexpected or sudden death only if autopsy proved CV-related. If there was doubt about the cause of death or there was no contact with the patient during the two years from study enrollment, that patient was excluded and not considered further.
Time frame: 2 years for each person qualified for the study
Diagnostic test: N-terminal pro-B-type natriuretic peptide (NT-proBNP)
N-terminal pro-B-type natriuretic peptide (NT-proBNP) \[fmol/ml\] concentration in the serum was analyzed by enzyme immunoassay using the Nt-proBNP kit from Biomedica, Slovakia.
Time frame: 3 years
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