Visual Scale as a Non-Invasive Method for Evaluation of Risk and Severity of Coronary Atherosclerosis.

Aim      To evaluate quantitative and qualitative characteristics of atherosclerotic plaques (ASP) in carotid arteries (CA) and femoral arteries (FA) and to use these data for developing a visual scale (VS) for noninvasive diagnosis and determination of severity of coronary atherosclerosis.Material and methods  This study included 216 patients (115 men and 101 women) aged 24 to 87 years (mean age, 61.5±10.73 years). All patients underwent coronary angiography (CAG) for detecting and determining severity of CA atherosclerosis and duplex scanning (DS) for detecting atherosclerosis of CA and FA.Results Analysis of ultrasound parameters of ASP in CA and FA showed that the maximal ASP height, moderate stenosis and maximal stenosis of the arterial bed had higher predictive values than other ultrasound parameters. These parameters were used for forming diagnostic complexes, on the basis of which two individual VSs for CA and FA were developed. Based on the high prognostic value of both scales, they were combined into one that was named VSCOMB. A ROC analysis determined cut-off points of the VSCOMB for diagnosis of CA atherosclerosis of various severity. VSCOMB scores  >4 indicated pronounced CA atherosclerosis with sensitivity of 86.1 % and specificity of 87.5 % whereas VSCOMB scores  ≤4 excluded it. Thus, VSCOMB score 0-1 indicated the absence of CA atherosclerosis; score 2-4 indicated the presence of subclinical CA atherosclerosis; and score >4 indicated severe CA atherosclerosis.Conclusion      A VSCOMB was developed that includes a set of ultrasound parameters for CA and FA and is useful for noninvasive diagnosis of CA atherosclerosis of various severity. Simple and convenient use of VSCOMB allows it to be used at the screening stage to detect subclinical CA atherosclerosis and to prevent its progression.

ISO 20387 biobanking standard. Analysis of requirements and experience of implementation.

Currently one of the most important problems facing biobanking specialists is the standardization of biobanks operation. Close attention is paid to this issue by international biobanking organizations, such as ISBER and BBMRI-ERIC, which develop regulatory documentation in this area. The article provides examples of standardization tools - implementation of the ISO 9001 quality standard and ISBER Best Practices. General information about the development, scope, and structure of the ISO 20387 standard is provided. The standard does not provide ready-made solutions and does not contain specific requirements for storage temperature or biosamples processing in biobanks, allowing each biobank to adapt its own management system to existing conditions and needs. The standard contains requirements for both the organization of the biobanking and the supporting processes - personnel competence; requirements for biological safety; infrastructure management, including equipment used by the biobank, environmental parameters that affect the storage of biomaterial. The standard contains requirements for the quality management system of biobank, as a necessary element of the organization of any biorepository. At the initiative of the Russian National Association of biobanks and biobanking specialists (NASBIO), development of the Russian standard GOST R ISO 20387 «Biotechnology. Collection and storage of biological samples in biobanks. General requirements¼ is included in the plan of the National Standardization Program for 2020 by order of Rosstandart No. 2612 of 11/01/2019. Implementing quality standards is a long and painstaking process that requires the involvement of all employees and certain resources. However, the effectiveness of strict compliance exceeds the cost of developing, implementing and maintaining management systems, as it significantly increases the confidence of researchers in the work of biobanks, guarantees high quality of biospecimens and associated data, and creates opportunities for cooperation, both at the national and international level, based on the application of common quality standards in the work.

[Prediction of Subclinical Coronary Atherosclerosis in Patients with High and Very High Cardiovascular Risk].

Objective To develop a diagnostic rule for detection of patients (pts) with high probability of subclinical atherosclerosis among those with high or very high cardiovascular (CV) risk.Materials and Methods This cross-sectional study enrolled 52 pts (32 men [62 %]), aged 40 to 65 years [mean age 54.6±8.0]) with high or very high CV risk (5-9 and ≥10 % by The Systematic Coronary Risk Estimation Scale [SCORE], respectively). All participants underwent cardiac computed tomography (CT) angiography and calcium scoring. Traditional risk factors (RFs) (family history of premature CVD, smoking, overweight / obesity and abdominal obesity, hypertension, type 2 diabetes mellitus, lipids parameters (total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides) and lipids-related markers (apolipoprotein A1, apolipoprotein B, ApoB / ApoA1 ratio), biomarkers of inflammation (high-sensitivity C-reactive protein [hs CRP], fibrinogen), indicator carbohydrate metabolism (glucose), ankle-brachial index, stress-test, carotid plaques according to ultrasound were evaluated in all pts. Psychological RFs were evaluated using Hospital Anxiety and Depression Scale and DS-14 for type D personality.Results All pts were divided into 2 groups according to the CT angiography results: pts in the main group (n=21) had any non-obstructive lesions or calcium score >0, pts in the control group (n=31) had intact coronary arteries. The groups did not differ in age or gender. 26 multiple linear logistic models for any subclinical atherosclerosis were developed based on obtained diagnostic features. Taking into account R-square = 0.344 (p=0.0008), the best fitting model was follows: subclinical coronary atherosclerosis= -1.576 + 0.234 x SCORE ≥5 % + 0.541 x hs CRP >2 g / l +0.015 x heart rate (bpm) +0.311 family history of premature CVD. The developed algorithm had sensitivity of 63 % and specificity of 80 %.Conclusion The created diagnostic model diagnostic model suggests the presence of subclinical coronary atherosclerosis in patients with high / very high CV risk with a high degree of probability. This easy-to-use method can be used in routine clinical practice to improve risk stratification and management choices in high-risk pts.

[An exploration of potential approaches to improve the diagnosis of subclinical atherosclerosis in patients with high cardiovascular risk].

PURPOSE: The search for optimal approaches to the diagnosis of subclinical atherosclerosis using a wide range of traditional and psychosocial risk factors (RFs), as well as clinical and instrumental diagnostic methods in patients (pts) with high or very high cardiovascular (CV) risk. METHODS: This cross-sectional study enrolled52 pts, aged 40 to 65 years with high or very high CV risk (5-9 and ≥10% by the Systematic Coronary Risk Estimation Scale [SCORE], respectively). All participants underwent cardiac computed tomography (CT)angiography and calcium scoring. Traditional RFs (family history of premature CVD, smoking, overweight/obesity and abdominal obesity, hypertension, type 2 diabetes mellitus, lipids parameters (total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides) and lipids-related markers (apolipoprotein A1, apolipoprotein B, ApoB/ApoA1 ratio), biomarkers of inflammation (high-sensitivity C-reactive protein [hs CRP], fibrinogen), indicator carbohydrate metabolism (glucose), ankle-brachial index, stress-test, carotid plaques according to ultrasound, arterial stiffness were evaluated in all pts. Psychological RFs were evaluated using Hospital Anxiety and Depression Scale and DS-14 for type D personality. RESULTS: All pts were divided into 2 groups according to the CT angiography results: pts in the main group (n=21) had any non-obstructive lesions or calcium score >0, pts in the control group (n=31) had intact coronary arteries. The groups did not differ in age or gender. It was found that patients with subclinical atherosclerosis significantly more often have a very high (≥10%) CV risk (42.9% vs.16.3%, p<0.05), a long (≥5 years) history of arterial hypertension (47.6% vs. 12.9% , p<0.01) and longer duration of antihypertensive therapy (61.9% vs. 29.0%, p<0.05), higher heart rate in rest (87. ± 14 vs. 77 ± 10 beats/min, p<0.01), increased arterial stiffness according to aortic pulse wave velocity (85.7% vs. 61.3%, p<0.05) and high level of hs-CRP (100% vs. 90.3%, p<0.05). CONCLUSION: Using in routine clinical practice of additional anamnestic (hypertension lasting ≥ 5 years and the intake of any antihypertensive drugs) and clinical-instrumental parameters (high heart rate in rest, hs CRP and arterial stiffness in pts with high and very high CV risk increases effectiveness of early detection of subclinical atherosclerosis.

[Quality management in biobank. World tendencies and experience of biobank of FSI «NMRC for preventive medicine¼ of the Ministry of healthcare of Russia.]

This paper provides several definitions of the term "biobank"; a list of standards developed by the International Organization for Standardization (ISO) applicable to the activities of biobanks; analyzes the legal and ethical requirements; the Russian legal framework in the field of biobanking, the best international practices and recommendations; describes the experience of the development and implementation of quality management systems according to ISO 9001 in Biobanks established in different countries, and the experience of the Bank of Biological Material of the National Medical Research Center for Preventive Medicine of the Ministry of Healthcare of Russia. The ISO 20387 Biobanking Standard, released in August 2018, combined the knowledge and experience of specialists from around the world and defined the general requirements that must be fulfilled by biobanks and repositories wishing to guarantee their customers the high quality of the preanalytical stage of scientific research, biological samples and associated data. In 2019, the Russian version of this standard is expected. Documents of the quality management system provide the reproducibility of activities on the main storage processes and facilitates the process of incorporating a new employee; conducting internal and external audits; Biobank knowledge management - continuous staff education. The introduction of an effective quality management system into biobank activity warrants the high quality of biological samples, the standardized pre-analytical stage, reliable, regulated long-term storage of biomaterial and related information for use in research purposes today and in future.

[Current best practices and biobanking recomedations.]

The biobank is a structure established with the goal of long-term responsible storage of biological samples and the associated data for their further use in scientific and clinical research. The objectives of biobanking are the creation of unified recommendations on: the planning of premises and the selection of equipment for storage; development of management methods and staff training; standardization of methods for the collection, shipping, processing and storage of biomaterial of various origins, as well as methods for quality control and validation of the applied methods; creation and use of databases of information accompanying biospecimens. The lack of common standards for conducting the preanalytical phase has been the cause of low accuracy and poor reproducibility of research results. To date, a large number of guidelines and best practices have been published that provide an answer to a wide range of problems in organizing the biobanking process. The article provides an overview of the most famous biobanking guidelines that can be used to solve various research problems. Biobanking in Russia is actively developing. Since 1996 there is a work on the legislative regulation of biobanking activities, as a result of which a number of regulatory documents have been issued. An important stage in the development of biobanking in Russia was the establishment of the "National Association of Biobanks and Biobanking Specialists" (NASBio) in 2018, which included representatives of medical and research institutions, commercial firms, and qualified specialists in the field of biobanking. One of the key tasks of NASBio is the adaptation and implementation of the best biobanking practices in Russian research institutes and centers. The use of modern guidelines and best practices on biobanking will lead to an increase in the quality of research and publications.

[Diagnostic Value of Biochemical Markers in Patients With Chronic Heart Failure With Reduced, Borderline and Preserved Left Ventricular Ejection Fraction].

PURPOSE OF THE STUDY: The study of the diagnostic value of biochemical markers of myocardial stress and inflammation in chronic heart failure (CHF) with different values of the ejection fraction (EF) of the left ventricle (LV). MATERIAL AND METHODS: The cross-sectional study included 105 patients aged 24 to 84 years (mean 58+/-14 years) with stable chronic heart failure I-II NYHA functional class classification. The causes of CHF were ischemic heart disease (IHD) in 33% of patients and arterial hypertension (AH) - 67%. All patients received medical treatment: angiotensin-converting enzyme (ACE) - 76%, -blockers - 72%, diuretics - 100%, statins - 80%. The control group consisted of 35 healthy volunteers. All subjects identified blood natriuretic peptide (NT-proBNP, ANP), adiponectin, galectin-3, pentraxins-3 and growth differentiation factor-15 (GDF-15) by enzyme immunoassay (ELISA). All surveyed performed transthoracic echocardiography (Echo). RESULTS: The blood NT-proBNP, ANP, galectin-3, pentraxins and GDF-3-15 in patients with chronic heart failure was significantly higher than in the control group (p<0.001 in all cases). In contrast, the level of adiponectin was significantly higher than in healthy individuals - 11.90 (11.39; 12.65) vs 7.73 (3.58; 8.86) ng/ml in patients with chronic heart failure (p<0.001). LVEF ranged from 30 to 55%, in 33% of patients it was >50%. Correlation analysis Spearman found strong correlations (p<0.001 for all markers) between LVEF and the content of all the biomarkers, while between the PV and the level of adiponectin is a positive correlation was found (r=0.862), and between the PV and the other biomarkers - reverse (r from -0.858 to -0.901). Multivariate linear regression analysis found the strongest correlation with the value of LVEF at pentraxin 3 and adiponectin. Subsequent ROC-analysis confirmed the diagnostic value of adiponectin in patients with heart failure and preserved ejection fraction. Thus, the level of adiponectin more than 8.3 ng/ml served as a prognostic factor for the presence of heart failure in patients with LVEF >50% with a sensitivity of 94.3% and a specificity of 92.9% (area under the curve 0.977; 95% confidence interval from 0.954 to 0.999; p<0.001). CONCLUSIONS: Among the 6 studied biomarkers of myocardial stress and inflammation only adiponectin has diagnostic significance in patients with heart failure and preserved ejection fraction.

[Combined markers of initial stages of coronary atherosclerosis]. / Kombinirovannye markery nachal'noi stadii ateroskleroza koronarnykh arterii.

Abnormalities in energy metabolism and endothelial dysfunction contribute to signaling processes associated with atherogenesis. The goal of our study was to develop diagnostic tests based on endothelial functional markers and adiponectin to differentiate early stages of coronary lesions during atherogenesis. The cohort included male and female patients from 25 to 86 years of age. All subjects underwent coronary angiography and severity of coronary lesions was quantified by the Gensini score that assigns points depending on location and extent of the lesions. We have estimated associations between the Gensini score and some known primary and secondary diagnostic parameters and have found that the ratio of serum levels of adiponectin to endothelin strongly correlates with severity of coronary lesions and can be used for differentiation of male patients lacking coronary atherosclerosis (despite symptoms of ischemic heart disease) from patients that have severe coronary lesions. Predictive power of adiponectin to endothelin ratio did not depend on drug therapy.

Direct labeling of serum proteins by fluorescent dye for antibody microarray.

Analysis of serum proteome by antibody microarray is used to identify novel biomarkers and to study signaling pathways including protein phosphorylation and protein-protein interactions. Labeling of serum proteins is important for optimal performance of the antibody microarray. Proper choice of fluorescent label and optimal concentration of protein loaded on the microarray ensure good quality of imaging that can be reliably scanned and processed by the software. We have optimized direct serum protein labeling using fluorescent dye Arrayit Green 540 (Arrayit Corporation, USA) for antibody microarray. Optimized procedure produces high quality images that can be readily scanned and used for statistical analysis of protein composition of the serum.

[Elevated Level of the Natriuretic Peptide Among Adult Population in Regions Participating in the ESSE-RF Study and Its Association With Cardiovascular Diseases and Risk Factors].

AIM: to study associations between elevated blood plasma concentration of N-terminal pro-brain natriuretic peptide (NT-proBNP), risk factors and cardiovascular diseases (CVD) in samples of adult population of Russian Federation (RF) aged 25-64 years. MATERIALS AND METHODS: We analyzed data of examination of representative samples of population of 5 regions of RF obtained within the framework of the multicenter ESSE-RF study (2012-2013). Number of examined subjects was 8 077 (3 176 men). Methods included use of standard questionnaire, measurements of height, body mass, blood pressure (BP), and plasma NT-proBNP level. The following CVD were included into analysis: arterial hypertension (AH), ischemic heart disease (IHD), atrial fibrillation (AF), and stroke. RESULTS: Women compared to men had higher NT-proBNT concentration was higher in women compared to men, in both genders it rose with age. Overall 17.9 % of examinees had elevated NT-proBNT levels (14.2 and 20.3 % among men and women, respectively). Elevated NTproBNP level was associated in men with age, myocardial infarction, angina pectoris, ischemic ECG changes, left ventricular hypertrophy, AF, bradycardia, smoking, in women with age, IHD, ischemic ECG changes, AF, bradycardia, heart rate ≥80 bpm, BP ≥160/95 mm Hg. CONCLUSION: In studied RF population elevated NT-proBNP level was significantly associated with gender, age, smoking, and CVD.

[Combination of Visual and Metabolic Markers in Assessment of Probability of Presence and Severity of Atherosclerosis of Coronary Arteries].

AIM: to assess possibilities of the use of biochemical markers combined with data of methods of imaging of arterial atherosclerotic lesions for evaluation of risk of presence and severity of coronary atherosclerosis. MATERIALS AND METHODS: We enrolled into this study patients (n=205, 136 men, 69 women, age 33-85 years, 94% on statin therapy) who underwent coronary angiography and carotid artery ultrasound dopplerography. Examination included determination of parameters of lipid profile and carbohydrate metabolism, markers of inflammation and metabolism of visceral adipose tissue. The severity of carotid artery atherosclerosis was estimated using mean common carotid artery intima-media thickness and atherosclerotic plaques presence. Severity of coronary atherosclerosis was evaluated using Gensini score. RESULTS: We found association between a number of biomarkers and severity of coronary artery involvement. Presence of coronary atherosclerosis (Gensini score >0) was associated with male sex, carotid artery stenosis >45%, and adiponectin level <8.0 pg/ml. Overt coronary atherosclerosis (Gensini score more or equal 35) significantly correlated with intima-media thickness more or equal 0.9 mm, C-reactive protein level >3.0 mg/l and adiponectin level <8.0 pg/ml.

[The application of lipoprint-system for analysis of sub-fractional spectrum of lipoproteins of blood serum.]

The lipoproteins of low and high density are presented by heterogeneous specter of particles differing by size, density, charge, composition and functional characteristics. The prevalence of small dense particles of lipoproteins of low and high density in blood plasma is associated with higher risk of development of coronary heart disease. The identification of subfractional spectrum of lipoproteins in clinical purposes is complicated because of requirement of expensive equipment and reagents and extended time of implementation. The lipoprint-system (Quantimetrix Lipoprint LDL/HDL System, USA) based on the vertical electrophoresis using 3% polyacrilamid gel, permits shortening time of sub-fractioning of lipoproteins up to three hours. In the spectrum of apoB-containing lipoproteins of very low density, intermediate density, C, B, A, lipoproteins of low density 1 and 2, small dense (lipoproteins of low density 3-7) are singled out. In the spectrum of lipoproteins of high density up to 10 sub-fractions associated in three groups and represented by large (lipoproteins of high density 1-3), intermediate (lipoproteins of high density 4-7) and small (lipoproteins of high density 8-10) particles are singled out. The article describes technique of identification of spectrum of particles of lipoproteins of low and high density in human blood serum. The conditions of implementation of experiments are presented. The advantages and limitations of technique are indicated. The number of examples of application of indices of sub-fractional spectrum of lipoproteins as additional markers of evaluation of aterogenity of lipid profile are presented. The conclusion is made concerning possibility of application of technique in clinical laboratory diagnostic.

[THE POSSIBILITIES OF APPLICATION OF TECHNOLOGY PROTEIN MICROARRAY (MICROCHIPS) FOR ANALYSIS OF PROTEIN COMPOSITION OF BLOOD SERUM].

The microchip technology represents convenient and relatively economic tool of analyzing specific biomarkers with the purpose to diagnose diseases, to evaluate effectiveness of therapy and to investigate signaling pathways. To analyze protein composition of blood serum certain types of finished microchips which were not applied previously on the territory of Russia. The detection from 2% to 5% out of matrix of chips depending on their variety was managed without preliminary depletion of serum (removal of proteins of major fractions). Hence, partial protein composition of blood serum can be analyzed with microchips even without preliminary removal of proteins of major fractions.

[Markers of visceral adipose tissue metabolic abnormalities in patients with coronary atherosclerosis in relation to the presence of type 2 diabetes mellitus]. / Markery narusheniya metabolizma vistseral'noi zhirovoi tkani u patsientov s koronarnym aterosklerozom v zavisimosti ot nalichiya sakharnogo diabeta 2-go tipa.

AIM: To assess the specific features of visceral adipose tissue metabolism in patients with coronary atherosclerosis, complicated or uncomplicated type 2 diabetes mellitus (T2DM). MATERIAL AND METHODS: A cohort of 429 patients (325 men and 104 women; mean age, 61.3±9.4 years) with coronary atherosclerosis who had been admitted to the National Research Centre for Preventive Medicine, Ministry of Health of Russia, to undergo coronarography (CG) and to receive high-tech treatments and met the criteria for being included in and excluded from this investigation, was examined. The Gensini scoring scale was used to estimate the magnitude of coronary atherosclerosis from CG RESULTS: Carotid artery duplex ultrasound scanning estimating the intima-media thickness was performed in 48% of the patients. DM was diagnosed from examination results (fasting plasma glucose ≥7.0 mmol/l and glycated hemoglobin >6.5%) and an endocrinologist's report. 94% of the patients took statins. RESULTS: Overall, 18% of the examinees had DM that was 2.5 times more common in the women than in the men (32.7 and 13.2%, respectively (p=0.000). The diabetic and non-diabetic patients showed no significant differences in age: 62.9±8.3 and 60.9±9.6 years, respectively (p=0.105). There were statistically significant differences in the magnitude of coronary artery atherosclerosis according to the Gensini scale in relation to the presence of T2DM; thus, the median Gensini score was 48 in the diabetic patients and 46 in the persons with no signs of the disease (Mann-Whitney test; p=0.03). Analysis of adipokine levels showed that the median leptin level was significantly higher than that in the male patients with T2DM than in the persons with no signs of the disease. In the patients with T2DM, the median adiponectin level turned out to be significantly lower in both men and women. CONCLUSION: The coronary atherosclerosis severity rated using the Gensini scale is shown to increase in the presence of T2DM. The probability of detecting obvious (>45%) carotid artery lesion is associated with the presence of DM in both men and women. The male patients with T2DM concurrent with coronary atherosclerosis are noted to have an elevated leptin level, but a lower adiponectin concentration was found in both the men and women.

Effects of alcohol on the major steps of reverse cholesterol transport.

The effects of moderate alcohol consumption on the capacity of blood sera to promote acceptance of cholesterol (C) from Fu5AH hepatoma cells, esterification of delivered free C, and transfer of produced cholesteryl esters to apolipoprotein (apo) B-containing lipoproteins have been studied. Twenty male subjects with relatively high (>50 mg/dl, n = 10) and low (<50 mg/dl, n = 10) high density lipoprotein (HDL) C levels consumed for eight weeks red grape wine (0.3 g ethanol/kg body mass per day). Alcohol consumption reduced total C and low density lipoprotein C levels in both groups of subjects. Low HDL C subjects showed an increase in HDL C, apo AI, apo AII, and lipoprotein (Lp) AI particle levels after alcohol consumption. Alcohol did not affect free C efflux from the cells. However, after the following period of substitution of alcohol with an isocaloric amount of red grape juice, cellular C efflux markedly reduced. While lecithin:cholesterol acyltransferase (LCAT) activity increased during alcohol consumption only in subjects with low HDL C, high HDL C subjects showed a significant decrease in cholesteryl ester transfer protein (CETP) activity. At the same time, alcohol consumption reduced the endogenous C esterification rate and increased the transfer of endogenous cholesteryl esters to apo B-containing lipoproteins in both groups. Thus, alcohol consumption in moderate doses enhanced the anti-atherogenicity of the serum lipoprotein spectrum, supporting more effective C efflux from peripheral cells and transport of accepted C to apo B-containing lipoproteins. The effects of alcohol on the reverse cholesterol transport depend on the initial HDL C level.