Clinical and Laboratory Biomarkers in Patients With Atherothrombotic Stroke

Study Overview
=================
Brief Summary
-----------------
Aim of the present study is to investigate molecular and clinical markers in patients with atherosclerotic carotid stenosis (ACAS) in the ischemic stroke acute phase.

Detailed Description
-----------------
Background: Laboratory biomarkers of atherosclerosis can be valuable in decision about operative treatment in patients with mild to severe atherosclerotic carotid stenosis (ACAS) and high stroke risk. However nowadays there are no established instruments for personalized atherothrombotic stroke diagnostics. Aim of the present study was to access atherosclerosis biomarkers serum levels in patients with ACAS during the ischemic stroke/ transient ischemic attack (TIA) acute phase. Main objective of the study To explore informative biomarkers to determine the risk of stroke in patients with significant ACAS. Secondary objectives To investigate the association between the degree of neurological and cognitive deficits , the features of the disease , the severity of brain lesions according to neuroimaging data with the concentration of lipoprotein -associated phospholipase A2 (LP-PL-A2), high sensitive C- reactive protein (hsCRP) , pregnancy-associated plasma protein A (PAPP-A), asymmetric dimethylarginine (ADMA), lipoprotein (a) in patients with atherothrombotic stroke. To find the most valuable laboratory biomarkers of atherosclerosis , to compare them with clinical and objective data to make decision about inclusion of these biomarkers in routine practice as a screening test of atherosclerotic plaque instability and for stroke risk prediction and in decision about operative treatment in patients with ACAS. Design and Methods A single-blind cross-sectional trial was performed to investigate laboratory biomarkers of atherosclerosis in patients with atherosclerotic stenosis of the internal carotid artery 50-99 % , and in healthy volunteers. Randomizing and blinding technique: laboratory scientist and statistician do not have information about the belonging of biomaterials patient to any of the groups studied . Examination of patients includes history taking, neurological examination, duplex ultrasound, mini mental score examination (MMSE), enzyme-linked immunosorbent assay (ELISA) performed atherosclerosis biomarkers serum level measurement (LP-PL-A2, PAPP-A, ADMA, hsCRP and blood lipid profile).

Conditions
-----------------
Carotid Artery Diseases, Stroke

Intervention / Treatment
-----------------
* Other: Biomarkers serum level measurement


Participation Criteria
=================
Eligibility Criteria
-----------------
Acute stroke Inclusion Criteria: 50-99% atherosclerotic carotid artery stenosis Acute atherothrombotic stroke/TIA (according to TOAST-criteria) within the first 3 days after vascular event Exclusion Criteria: risk factors of non-atherothrombotic stroke (according to TOAST-criteria) cancer exacerbation of decompensated chronic diseases infections acute cardiovascular diseases large operation during 1 month before enrollment Cortexin, Actovegin, Cerebrolysin, Erythropoetin administration during 1 week before enrollment. Stable ACAS Inclusion Criteria: 50-99% atherosclerotic carotid artery stenosis no history of vascular events during one month before enrollment Exclusion Criteria: risk factors of non-atherothrombotic stroke (according to TOAST-criteria) cancer exacerbation of decompensated chronic diseases infections acute cardiovascular diseases large operation during 1 month before enrollment Cortexin, Actovegin, Cerebrolysin, Erythropoetin administration during 1 week before enrollment. Control group Inclusion Criteria: intima media thickness less then 1mm no history of stroke/TIAs Exclusion Criteria: risk factors of non-atherothrombotic stroke (according to TOAST-criteria) cancer exacerbation of decompensated chronic diseases infections acute cardiovascular diseases large operation during 1 month before enrollment Cortexin, Actovegin, Cerebrolysin, Erythropoetin administration during 1 week before enrollment.

Ages Eligible for Study
-----------------
Minimum Age: 50 Years
Maximum Age: 80 Years

Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
Accepts Healthy Volunteers

Study Plan
=================
How is the study designed?
-----------------

Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Acute stroke<br>Patients in the acute phase of atherothrombotic stroke or TIAs with 50-99% ACAS within the first 3 days af vascular event. Interventions to be administered: Biomarkers serum level measurement, history taking, neurological examination, duplex ultrasound, MMSE, National Institutes of Health Stroke Scale (NIHSS) | Other: Biomarkers serum level measurement<br>* Biomarker serum level measurement was performed with ELISA using following kits: Lp-PL-A2 ELISA- Cloud-CloneCorp. (USA); PAPP-A ELISA- IBL (Germany); Lp (a) ELISA Kit-AssayPro (USA ), ADMA ELISA Kit- ImmunDiagnostik ( Germany); hsCRP ELISA- Biomerica (Germany); according to the manufacturer's instructions . Absorbance of standards and samples was measured with a microplate reader Bio-Tek (USA) at a wavelength specified by the kit manufacturer . The calculation of the determined biomarkers concentration was performed using the software SOFTmaxPRO.<br>|
| Stable carotid artery stenosis<br>Patients with 50-99% ACAS without history of vascular events during one month before enrollment. Interventions to be administered: Biomarkers serum level measurement, history taking, neurological examination, duplex ultrasound, MMSE | Other: Biomarkers serum level measurement<br>* Biomarker serum level measurement was performed with ELISA using following kits: Lp-PL-A2 ELISA- Cloud-CloneCorp. (USA); PAPP-A ELISA- IBL (Germany); Lp (a) ELISA Kit-AssayPro (USA ), ADMA ELISA Kit- ImmunDiagnostik ( Germany); hsCRP ELISA- Biomerica (Germany); according to the manufacturer's instructions . Absorbance of standards and samples was measured with a microplate reader Bio-Tek (USA) at a wavelength specified by the kit manufacturer . The calculation of the determined biomarkers concentration was performed using the software SOFTmaxPRO.<br>|
| Control group<br>Healthy volunteers without ACAS Interventions to be administered:Biomarkers serum level measurement, history taking, neurological examination, duplex ultrasound, MMSE | Other: Biomarkers serum level measurement<br>* Biomarker serum level measurement was performed with ELISA using following kits: Lp-PL-A2 ELISA- Cloud-CloneCorp. (USA); PAPP-A ELISA- IBL (Germany); Lp (a) ELISA Kit-AssayPro (USA ), ADMA ELISA Kit- ImmunDiagnostik ( Germany); hsCRP ELISA- Biomerica (Germany); according to the manufacturer's instructions . Absorbance of standards and samples was measured with a microplate reader Bio-Tek (USA) at a wavelength specified by the kit manufacturer . The calculation of the determined biomarkers concentration was performed using the software SOFTmaxPRO.<br>|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Mean concentration of Lipoprotein-associated Phospholipase A2 (LP-PL-A2) |  | 0 month | 
| Mean concentration of Pregnancy-associated Plasma Protein A (PAPP A) |  | 0 month | 
| Mean concentration of Asymmetric Dimethylarginine (ADMA) |  | 0 month | 
| Mean concentration of highsensitivity C-reactive Protein (hsCRP) |  | 0 month | 
| Mean concentration of Lipprotein (a) |  | 0 month | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
atherosclerosis, biomarkers, carotid artery stenosis, ischemic stroke.