Using the method of dielectrophoresis an optical cell detection system in an inhomogeneous alternating electric field, the electrical and viscoelastic parameters of red blood cells in the frequency range from 5 × 104 to 1 × 106 MHz have been studied in 129 patients with colorectal cancer (CRC) of various stages of the disease, 25 people with adenomatous polyps, and 35 people in the comparison group. It has been shown that the values of the physical characteristics of red blood cells significantly differ both when comparing the control groups and the patients with adenomatous polyps and the patients with early stages of CRC and when comparing the patients with different stages of the oncological process with each other. The construction of SHAP graphs based on the electrical and viscoelastic parameters of red blood cells has allowed us to achieve 100% diagnostic accuracy in distinguishing patients with adenomatous polyps and early stages of CRC from healthy individuals, 98.7% accuracy—in differentiating the patients with polyps from the patients with early stages of CRC, and 91.9% accuracy—in distinguishing early and late stages of CRC. The results obtained demonstrate the high potential of the proposed method for screening and diagnosing CRC.

С помощью метода диэлектрофореза в неоднородном переменном электрическом поле при использовании оптической системы детекции клеток проведено исследование электрических и вязкоупругих параметров эритроцитов в частотном диапазоне от 5·104 до 1·106 MHz у 129 больных с колоректальным раком (КРР) различных стадий развития заболевания, 25 человек с аденоматозными полипами и 35 лиц группы сравнения. Показано, что величины физических характеристик эритроцитов значимо различаются как при сопоставлении групп контроля и пациентов с аденоматозными полипами и больных с ранними стадиями КРР, так и при сравнении пациентов с различными стадиями онкологического процесса между собой. Построение SHAP-графиков на основе электрических и вязкоупругих параметров эритроцитов позволило достичь 100% диагностической точности при различении пациентов с аденоматозными полипами, ранними стадиями КРР от здоровых лиц, 98.7% точности --- при дифференцировании пациентов с полипами от больных с ранними стадиями КРР, 91.9% точности --- при различении ранних и поздних стадий КРР. Полученные результаты демонстрируют высокий потенциал предложенного метода для скрининга и диагностики КРР. Ключевые слова: диэлектрофорез, эритроциты, электрические и вязкоупругие параметры, диагностика, колоректальный рак, аденоматозные полипы, SHAP-графики.

The dielectric properties of the erythrocyte were studied. Experimental methods and results of measuring the dielectric properties of individual human erythrocyte are presented. The method is theoretically justified. It is found that values of the complex permittivity of the erythrocyte, electrical capacitance, dielectric loss tangent remain almost constant despite significant changes in conductivity and the content of NaCl in the cell suspension. These values reflect the stability of the studied parameters of erythrocytes under conditions of changes within a wide range of the environment in which the cells are located. Complex cell permittivity and cell dielectric loss tangent are promising parameters for objective diagnostics of human diseases. The introduction of measurements of cell permittivity and cell dielectric loss tangent into medical practice will likely allow us to characterize the process of individual cell self-regulation in more detail.

The aim of the work is to study the differences in changes in the viscoelastic parameters of erythrocytes (amplitude of deformation, summarized viscosity and summarized rigidity) in the dynamics of interaction with ethanol in vitro in patients with non-alcoholic (NAFLD) and alcoholic (AFLD) fatty liver disease for use in differential diagnostics. Materials and methods. 50 men were examined, 24 - with NAFLD (46.5±1.5 years), 26 - with AFLD (48.6±1.2 years). The study of viscoelastic parameters of erythrocytes was carried out by dielectrophoresis with an electro-optical cell detection system. The levels of erythrocytes were evaluated before and after in vitro exposure for 300 s with 10 μl of a 0.02% ethanol solution in the dynamics of the experiment. Results. In erythrocytes of patients with NAFLD after exposure with ethanol, a decrease in strain amplitude was revealed against the background of an increase in summarized viscosity and summarized rigidity, on the contrary, in erythrocytes in patients with AFLD, an increase in the ability to deform was observed with a decrease in generalized viscosity and rigidity (p <0.01-0.05). Inverse reaction of cells to ethanol exposure allowed to distinguish patients with fatty liver disease of various origins with sensitivity of 87.5%, specificity of 96.2%, predictive value of positive - 95.4% and negative - 89.3%, as well as an accuracy of 92%. The proposed method for distinguishing fatty liver disease of various origins is promising due to its simplicity, high throughput and low cost.

The aim of the work is to study the differences in changes in the viscoelastic parameters of erythrocytes (amplitude of deformation, summarized viscosity and summarized rigidity) in the dynamics of interaction with ethanol in vitro in patients with non-alcoholic (NAFLD) and alcoholic (AFLD) fatty liver disease for use in differential diagnostics. Materials and methods. 50 men were examined, 24 - with NAFLD (46.5±1.5 years), 26 - with AFLD (48.6±1.2 years). The study of viscoelastic parameters of erythrocytes was carried out by dielectrophoresis with an electro-optical cell detection system. The levels of erythrocytes were evaluated before and after in vitro exposure for 300 s with 10 μl of a 0.02% ethanol solution in the dynamics of the experiment. Results. In erythrocytes of patients with NAFLD after exposure with ethanol, a decrease in strain amplitude was revealed against the background of an increase in summarized viscosity and summarized rigidity, on the contrary, in erythrocytes in patients with AFLD, an increase in the ability to deform was observed with a decrease in generalized viscosity and rigidity (p <0.01-0.05). Inverse reaction of cells to ethanol exposure allowed to distinguish patients with fatty liver disease of various origins with sensitivity of 87.5%, specificity of 96.2%, predictive value of positive - 95.4% and negative - 89.3%, as well as an accuracy of 92%. The proposed method for distinguishing fatty liver disease of various origins is promising due to its simplicity, high throughput and low cost.

AbstractThe possibilities of using electrical and viscoelastic parameters of erythrocytes in patients with colorectal cancer (CRC) of different stages were demonstrated using optical cell detection system based on the dielectrophoresis method. The deformation amplitude, portion of deformed cells, generalized viscosity, rigidity, equilibrium frequency position, and electrical conductivity were the most significant parameters for determining patients with CRC, as well as early and late stages, included in the created model of a riskometer for the disease diagnosis. The created diagnostic “panels” with a high diagnostic accuracy, sensitivity, and specificity allowed to one determine patients with CRC (specificity 0.91, sensitivity 0.96), as well as early and late stages of the disease (specificity 0.64, sensitivity 0.86).

The possibilities of using electrical and viscoelastic parameters of erythrocytes in patients with colorectal cancer (CRC) of different stages were demonstrated using optical cell detection system based on the dielectrophoresis method. The deformation amplitude, portion of deformed cells, generalized viscosity, rigidity, equilibrium frequency position, and electrical conductivity were the most significant parameters for determining patients with CRC, as well as early and late stages, included in the created model of a riskometer for the disease diagnosis. The created diagnostic panels with a high diagnostic accuracy, sensitivity, and specificity allowed to one determine patients with CRC (specificity 0.91, sensitivity 0.96), as well as early and late stages of the disease (specificity 0.64, sensitivity 0.86).

The possible mechanisms for the development of aspirin resistance, platelet involvement in thrombus formation, inflammation and atherogenesisare presented. The presence of associations between aspirin resistance and the risk of thrombotic complications in people not using aspirin and the possibility of using aspirin resistance to assess the individual risk of thromboembolic complications in 492 patients (mean age 56.2 ± 0.9 years) with arterial hypertension I - Stage II and without ithas been researched. The parameters of platelet aggregation were studied in subgroups of patients different in the risk of developing venous thrombo-embolic complications by reaction to the cyclooxygenase blocker. It has been revealed that the conducted antiplatelet therapy is effective in 50-60 % of persons with cardiovascular diseases; ~ 40 % of patients show signs of relative resistance to antiplatelet agents. Resistance to therapeutic doses of antiplatelet agents is multifactorial, however, the presence of a shunt between thromboxane synthetase and lipoxygenase plays a role in it. It is likely that the influence of inflammation on the development of aspirin resistance through an increase in the synthesis of 12-NETU, thromboxane and leukotrienes. Aspirin resistance is a risk factor for the development of venous thrombotic complications, regardless of the use of antiaggregant therapy. It was established that the most accurate prediction of the result according to the severity of hemorheological disorders (distinguishing between moderate and severe disorders from the lungs) is provided by such indicators of platelet aggregation as the coefficient K ((ADF5 % - ADP 5 % + Asp) - 100 / ADF 5 %) (AUC 0.995) and ADP 5 % (AUC 0.713). Changes in platelet aggregation in response to aspirin in an in vitro test can be used to identify individuals with a high risk of thrombotic complications, and to assess individual risk (specificity 92.9-98.2 %, sensitivity 83.7-93 %).

ВОЗМОЖНОСТИ ОЦЕНКИ СТЕПЕНИ ТЯЖЕСТИ ГЕМОРЕОЛОГИЧЕСКИХ НАРУШЕНИЙ У ЛИЦ С АРТЕРИАЛЬНОЙ ГИПЕРТЕНЗИЕЙ 1

We present the experimental results of measurements of the masses of a separately observed erythrocyte and a latex particle placed in a variable electric field. We show that the experimentally measured and theoretically computed masses of reference latex particles are in good agreement.

A new state standard of biological cell polarizability based on micron-size latex particles has been developed. As a standard material, it is suggested to use polystyrene. Values of the polarizability calculated for erythrocytes and values of the polarizability of micron-size spherical latex particles measured with measuring-computing complexes agree within the limits of satisfactory relative error. The Standard allows one the unit of polarizability measurements [m3] to be assigned to cells and erythrocytes for the needs of medicine.

The possible mechanisms for the development of aspirin resistance, platelet involvement in thrombus formation, inflammation and atherogenesisare presented. The presence of associations between aspirin resistance and the risk of thrombotic complications in people not using aspirin and the possibility of using aspirin resistance to assess the individual risk of thromboembolic complications in 492 patients (mean age 56.2 ± 0.9 years) with arterial hypertension I - Stage II and without ithas been researched. The parameters of platelet aggregation were studied in subgroups of patients different in the risk of developing venous thrombo-embolic complications by reaction to the cyclooxygenase blocker. It has been revealed that the conducted antiplatelet therapy is effective in 50-60 % of persons with cardiovascular diseases; ~ 40 % of patients show signs of relative resistance to antiplatelet agents. Resistance to therapeutic doses of antiplatelet agents is multifactorial, however, the presence of a shunt between thromboxane synthetase and lipoxygenase plays a role in it. It is likely that the influence of inflammation on the development of aspirin resistance through an increase in the synthesis of 12-NETU, thromboxane and leukotrienes. Aspirin resistance is a risk factor for the development of venous thrombotic complications, regardless of the use of antiaggregant therapy. It was established that the most accurate prediction of the result according to the severity of hemorheological disorders (distinguishing between moderate and severe disorders from the lungs) is provided by such indicators of platelet aggregation as the coefficient K ((ADF5 % - ADP 5 % + Asp) - 100 / ADF 5 %) (AUC 0.995) and ADP 5 % (AUC 0.713). Changes in platelet aggregation in response to aspirin in an in vitro test can be used to identify individuals with a high risk of thrombotic complications, and to assess individual risk (specificity 92.9-98.2 %, sensitivity 83.7-93 %).

УДК 616.379-008.64-71ВОЗМОЖНОСТИ ДИАГНОСТИКИ НАРУШЕНИЙ РЕОЛОГИИ У БОЛЬНЫХ САХАРНЫМ ДИАБЕТОМ 2 ТИПА Кручинина М.В. 1 , Громов А.А. 1 , Паруликова М.В. 1 , Головин М.Г. 1 , Баум В.А. 1 , Генералов В.М. 2 , Генералов К.В. 2 , Кручинин В.Н. 3 1 Научно-исследовательский институт терапии и профилактической медицины -филиал Федерального государственного бюджетного научного учреждения «Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук», Новосибирск

The experimental method is described and the measurement results of the value of polarizability of erythrocytes and latex particles in a non-uniform alternating electric field are presented. The results show that the values of experimentally measured and theoretically calculated polarizablity values of latex particles are in good agreement.

Theoretical analysis of the relative error of measurement of polarizability of cells using the dielectrophoresis method is described. Information about the magnitude of error ensures objective evaluation of the disease dynamics and the effectiveness of treatment.

UNLABELLED:The aim of this work was to assess the potential of some physical and chemical methods for studying erythrocytes and blood serum in gastroenterological practice by the example of colorectal cancer (CC).MATERIALS AND METHODS:A total of 26 persons with various stages of colorectal cancer and 16 healthy (control group) were examined. Parameters of erythrocytes and blood serum were investigated by light microscopy, dielectrophoresis in a non-uniform alternating electric field (DEF in NUAEF), terahertz spectroscopy, ellipsometry, Raman-spectroscopy.RESULTS:Polymorphism of erythrocytes, rigidity, viscosity, indexes of aggregation and destruction were significantly higher in patients with CC and polarizability, amplitude of erythrocyte deformation in NUAEF being lower than those in the controls. The study of erythrocytes by terahertz spectroscopy revealed the low levels of amplitude transmittance over the whole frequency range in CC patients compared to the controls. The increasing of refractive index, degree of heterogeneity of thin films obtained from the serum in CC patients were observed in considering the ellipsometric parameters. We found a significant increasing of the concentration of antigens to CD24 at the early stage of the disease. The areas of some peaks in Raman spectra were significantly lower in patients with CC compared to the healthy ones, it is possible due to a carotin deficiency. Most of the studied parameters were correlated with the stage of the disease. A set of optical methods for studying blood serum compared with those of histology and radiological methods of diagnosis showed their high sensitivity and specificity, positive and negative predictive value (80 % and above).CONCLUSION:The obtained results of the pilot study demonstrate the prospects of using physical and chemical methods of research of erythrocytes and blood serum for early diagnosis, stage of disease and monitoring the effectiveness of treatment of CC.

The features of the electrical and viscoelastic parameters of red blood cells obtained by dielectrophoresis in patients with chronic hepatitis B in depending on the level of viral load were investigated. It has been found that the summerized rigidity, viscosity, conductivity, index of aggregation and destruction, the relative polarizability significantly increased and polarizability at all frequencies, the velocity of cells to the electrodes, electric capacitance of membranes — decreased as ascending viremia. A marked change in the parameters of red blood cells in patients with high viral load, likely, reflects the expressed degree of disadaptation of erythrocytes. Correlation of electrical and viscoelastic parameters erythrocytes with the biochemical indices reflecting the level of dysproteinemia, cytolysis, immune-inflammatory syndrome have been installed. The data obtained allow us to supplement understanding of the pathogenesis of systemic manifestations of virus hepatitis B and association of high viral load with a significant change in the viscoelastic and electrical properties of red blood cells.