Laboratory of Biophysical Basis of Pathology



Head of the Laboratory
Azizova, Ofelia Ahatovna
Doctor of Biological Sciences, Professor

Aseichev, A.B., Senior Researcher, Ph.D.
Beckman, E.M., Senior Researcher, Ph.D.
Piryazev, A.P., Senior Researcher, Ph.D.
Dudnik, L.B., Senior Researcher, D.Sc.
Scheglovitova, O.N., Junior Researcher, D.Sc.
Shvachko, A.G., Junior Researcher
Gubareva, E.V., Laboratory Assistant
Akimova, N.I., Laboratory Assistant
Salimov, E.L., Laboratory Assistant
Osokin, A.S., Engineer.

Topics of interest

Topics of interest

The laboratory aims to study the role of oxidative stress in cardiovascular diseases. Three main lines of research are conducted in our laboratory:

"Studies on the influence of oxidatively modified lipids and proteins on functions of the blood and blood vessel cells and on thrombosis"

Problems that are to be solved:
1. Investigation of the effects of oxidatively modified albumin on the functions of blood cells and endothelial cells as well as on the blood clotting system.
2. Investigation of the dynamics of changes in the size of oxidatively modified fibrinogen, through using various models of free radical oxidation.


  • Turbidimetry
  • Chemiluminescence
  • EIA
  • Cytofluorometry
  • Coagulometry
  • Dynamic light scattering method
  • The method for studying Co2+-binding capacity of serum, which binding capacity for cobalt ions which allows evaluating the extent of ischemia of organs and the body as a whole

Responsible executives:
Aseichev, A.B., Beckman, E.M., Piryazev, A.P., Scheglovitova, O.N., Shvachko, A.G.

"Development of the method for evaluating disease severity, prognosing disease progression, and assessing treatment effectiveness for patients with cardiovascular diseases, through detection of biomarkers of oxidative stress."

For this purpose, we investigate the relationship between clinical biochemical parameters in patients with cardiovascular diseases and parameters characteristic for oxidative stress.

Analysis of the following markers of oxidative stress is carrying out:

I. Oxidized lipids:
1) Primary products of lipid peroxidation:
- hydroperoxides, diene conjugates.
2) Secondary products of lipid peroxidation:
- TBA-reactive products, prostaglandin F2-alpha (the work conducted together with the Genetic Engineering Laboratory)
- Induced chemiluminescence

II. Protein oxidation products:
- Carbonyl products
- Dityrosine
- SH-groups
- Advanced oxidation protein products (AOPP)
- Oxidatively modified fibrinogen
- Ischemia modified albumin (detected with a Co(II)-albumin binding assay)
- Modification of the binding properties of albumin (tested with the fluorescent probe K-35)

III. Measurement of levels and activity of pro-oxidative (pro-ox) and anti-oxidative (anti-ox) enzymes in blood plasma.
- Measurement of the activity of anti-oxidative enzymes, such as superoxide dismutase, glutathione peroxidase and catalase, in blood plasma
- Measurement of the activity of the pro-oxidative enzyme myeloperoxidase which characterizes the extent of oxidative stress
- Measurement of plasma levels of pro- and anti-oxidative enzymes by using EIA

"Development of the method for estimating the concentration of gold nanoparticles (NP) in blood and examination of the effect of nanoparticles on blood cells and blood clotting system"

The following studies are being carried out in this direction:

I. Investigation of the possibility to detect gold nanoparticles in different media (water, buffers, deluted and whole blood plasma, whole blood):

1) Examination of the dependence of the intensity of light absorption and scattering on the gold nanoparticle concentration
2) Determination of the particle size range suitable for measuring by light absorption and scattering methods
3) Estimation of the upper and lower limits for the possibility to measure gold nanoparticles
4) Development of the approach to measuring gold nanoparticles in blood plasma by the use of optical spectral methods for diluted plasma

II. Investigation of the influence of gold NP on the functions of blood cells such as leukocytes, erythrocytes, and platelets

Problems that are to be solved:

  1. Investigation of the effects of gold NP on the production of reactive oxygen species (ROS) by leukocytes
  2. Investigation of the effects of gold NP on osmotic and tert-butyl hydroperoxide (tert-BuOOH)-induced hemolysis of erythrocytes and on erythrocyte aggregation
  3. Investigation of spontaneous and ADP-induced platelet aggregation in the presence of gold NP

Methods of investigation.

  • The detection of NP absorption spectra on the Beckman DU-530 spectrophotometer
  •  The determination of nanoparticle size by measuring hydrodynamic size (diameter) of NP using the dynamic light scattering technique (the ZetasizerNano ZS (Malvern, UK))

Responsible executives:
Beckman, E.M., Aseichev, A.B., Piryazev, A.P., Dudnik, L.B., Shvachko, A.G.

Major scientific achievements

Major scientific achievements

Beginning in 1985, our laboratory’s community has conducted research on the problem of elucidating the role of free radical processes in the pathogenesis of atherosclerosis. In view of this, we undertook to investigate the alterations of the structure and function of blood lipoproteins and cells upon the development of atherosclerosis, and to examine the effects of oxidized lipoproteins on the functions of blood cells. Oxidized low-density lipoproteins were found to induce aggregation of platelets and erythrocytes, to activate leukocytes, to cause a decrease in lymphocyte activity, to activate clotting, and to inhibit fibrinolysis.
Over the years we accumulated a great experience in investigation of the oxidizability of blood plasma and the relationship between the oxidizability parameters and the degree of severity of ischemic heart disease (IHD) and blood hypertension. We gained experience to investigate the issue on the significance of free radicals in different cardiovascular pathologies and to estimate the level of free radical oxidation by measuring lipid peroxidation indices, such as primary (hydroperoxides) and secondary (TBA-reactive products and diene conjugates) products, as well as a protein oxidation index such as the carbonyl group content.

The main results of this research are the following:

  1. The correlation was shown between the level of oxidative stress (OS) markers and the degree of severity of IHD and the presence of accompanying symptoms (diabetes, obesity, blood hypertension). The high activity of free radical processes was demonstrated to correlate with parameters of electrical activity of ischemic myocardium.
  2. The level of OS markers was shown to be increased in patients with arrhythmia. The number of persons with an alteration of cardiac rhythm in the group of patients with high OS was 45 % greater than that in the group of patients that had lower degree of OS.
  3. The data obtained suggest that values of OS markers in combination with values of electrographic parameters may be used for risk stratification of patients and for decision making on treatment with antioxidant drugs as well as for monitoring the efficacy of antioxidant therapy. Thresholds of these values were defined.
  4. The exercise tolerance in chronic IHD patients whose levels of OS markers were higher than threshold values was found to decrease sharply in 1 – 3 years.
  5. OS markers were examined in patients with acute coronary syndrome (ACS). It was observed that in patients with the OS marker levels exceeding threshold values, 90 % of cases of heart attack were followed, in 5 - 7 days, by the alteration by heart rhythm. Also, such patients had 2 – 4 times more unfavorable long-term outcomes (repeated hospitalization for ischemic heart disease aggravation, heart attack, or arhythmic complications) compared to patients with low levels of OS markers. Knowledge of these threshold values could be used to predict long-term adverse cardiovascular events after acute myocardial infarction and to choose an appropriate preventive treatment.
  6. Levels of OS markers in patients with cerebral ischemia were found to be considerably higher (by 40 %) compared to healthy individuals. Threshold values of OS markers were defined. OS levels characterized by marker values higher than the defined threshold values were associated with an unfavorable progression of chronic cerebral ischemia. These threshold values could be used for prediction of an adverse course of the disease and for well-timed choice of adequate treatment to prevent stroke.
  7. The data on OS markers were obtained for patients with obliterative atherosclerosis of lower extremities (OALE) and IHD. The values of OS markers were higher in patients with OALE and correlated with the degree of IHD. Threshold values were defined, which could allow a risk assessment of an adverse course of the disease in patients with OALE and IHD.

For the last decade, our laboratory has investigated the influence of oxidized fibrinogen on the activity of blood cells (such as leukocytes, platelets, and erythrocytes) as well as on blood clotting and fibrinolysis. Oxidatively modified fibrinogen was found to increase platelet aggregability and the generation of reactive oxygen species by leukocytes. Further analysis using inhibitors indicated that the mechanism of platelet activation involved phospholipase C, protein kinase C and tyrosine kinase. It was found that low doses of fibrinogen added to blood or plasma caused activation of processes which are similar to pathways of disseminated intravascular coagulation.
We developed, for the first time, and proved a method to detect gold nanoparticles (NP) directly in blood plasma through light scattering and absorption spectroscopy. We proposed a new approach for detecting gold NP in plasma. The protocol includes the dilution of plasma samples with phosphate buffered saline (PBS). Based on experimental data, we validated the 1/100 dilution of plasma in order to obtain optimal results when using light scattering spectroscopy. Less diluted samples (up to 10 % plasma) are needed to allow the detection of gold NP by means of light absorption. So, the proposed method consists in the quantitative detection of gold NP in blood plasma through light scattering and absorption spectroscopy and constructing calibration graphs against known NP concentrations. We ascertained that light absorption spectroscopy may be applied to the detection of relatively small NP, while for big NP (30 nm or greater in diameter), light scattering spectroscopy is more suitable.
Our preliminary results concerning the effect of gold NP on blood cells, such as leukocytes, erythrocytes and platelets, suggest that the studied gold NP are biocompatible and may be used for targeted delivery of drugs and photodynamic therapy.

Our research results are presented in more than 50 publications in scientific journals. Three patents were obtained.



  • Spectrophotometer СФ-121
  • Spectrophotometer Becman DU-530
  • Microplate photometer Multickan Ascent
  • Microplate photometer-fluorimeter ФФМ-1
  • Сentrifuges OПН -3М
  • Analitical balance Ohaus
  • Distilling apparatus
  • Chemiluminometer Биотокс-7
  • Spectrofluorimeter «Percin Elmer» LS-50
  • Aggrecation analyser Биола LA-220 and LA-250



Piryazev AP, Azizova OA, Aseichev AV, Dudnik LB, Sergienko VI. Effect of gold nanoparticles on production of reactive oxygen species by human peripheral blood leukocytes stimulated with opsonized zymosan. Bull Exp Biol Med. 2013 Nov;156(1):101-3.

Aseychev AV, Azizova OA, Beckman EM, Dudnik LB, Sergienko VI. Effect of gold nanoparticles coated with plasma components on ADP-induced platelet aggregation. Bull Exp Biol Med. 2013 Sep;155(5):685-8.

Azizova OA, Solov'eva EY, Aseichev AV, Baranova OA, Bekman EM, Karneev AN, Mironova OM, Manevsky AP, Ivanokov AN, Fedin AI, Sergienko VI. [The association between oxidative stress markers and clinical course of chronic cerebral ischemia]. Zh Nevrol Psikhiatr Im S S Korsakova. 2013;113(9 Pt 2):21-7. Russian.

Aseychev AV, Azizova OA, Beckman EM, Skotnikova OI, Piryazev AP, Dobretsov GE. Studies of oxidant-induced changes in albumin transport function with a fluorescent probe K-35. Metal-catalyzed oxidation. Bull Exp Biol Med. 2012 Aug;153(4):463-7. English, Russian.

Azizova OA, Aseychev AV, Beckman EM, Moskvina SN, Skotnikova OI, Smolina NV, Gryzunov YA, Dobretsov GE. Studies of oxidant-induced changes in albumin transport function with a fluorescent probe k-35. Effect of hypochlorite. Bull Exp Biol Med. 2012 Apr;152(6):712-6. English, Russian.

Koroleva OS, Pushkov AA, Blagodatskikh KA, Baranova OA, Azizova OA, Nosikov VV, Reznichenko NE, Zateĭshchikov DA. [Association of a polymorphic marker Trp719Arg of KIF6 gene with effects of atorvastatin and simvastatin in patients with early ischemic heart disease]. Kardiologiia. 2011;51(9):4-12. Russian.

Aseĭchev AV, Azizova OA, Shcheglovitova ON, Skliankina NN, Borisenko GG. [The influence of oxidized fibrinogen on apoptosis of endothelial cells]. Biomed Khim. 2011 Mar-Apr;57(2):210-8. Russian.

Belenkov IuN, Privalova EV, Chekneva IS, Zheleznych EA, Khiazeva LV, Azizova OA, Aseĭchev AV, Baranova OA, Shvachko AG. [Comparative analysis of antioxidant activity of nebivolol in patients with chronic heart failure with and without concomitant type 2 diabetes]. Kardiologiia. 2011;51(1):5-10. Russian.

Piryazev AP, Aseichev AV, Azizova OA. Effect of oxidation-modified fibrinogen on the formation and lysis of fibrin clot in the plasma. Bull Exp Biol Med. 2009 Dec;148(6):881-5.

Azizova OA, Solov'eva EIu, Mironova OP, Bekman EM, Baranova OA, Moskvina SN, Fedin AI. [Relationship between clinical characteristics and primary and secondary products of lipid and protein peroxidation]. Vestn Ross Akad Med Nauk. 2010;(1):18-23. Russian.

Azizova OA, Shvachko AG, Aseichev AV. Effect of iron ions on functional activity of thrombin. Bull Exp Biol Med. 2009 Nov;148(5):776-9.

Azizova OA, Sergienko VI, Syrkin AL, Ivanov GG, Aseĭchev AV, Lopukhin IuM. [Clinical and prognostic significance of free radical processes in patients with coronary heart disease]. Vestn Ross Akad Med Nauk. 2009;(10):32-40. Review. Russian.

Aseychev AV, Azizova OA, Shulenina LV, Piryazev AP. Effect of oxidized fibrinogen on aggregation of activated platelets and neutrophils. Bull Exp Biol Med. 2009 Mar;147(3):312-8.

Azizova OA, Piryazev AP, Aseychev AV, Shvachko AG. Oxidative modification of fibrinogen inhibits its transformation into fibrin under the effect of thrombin. Bull Exp Biol Med. 2009 Feb;147(2):201-3.

Solov'eva EIu, Mironova OP, Baranova OA, Bekman EM, Aseĭchev AV, Fedin AI, Azizova OA. [The free-radical processes and antioxidant therapy in brain ischemia]. Zh Nevrol Psikhiatr Im S S Korsakova. 2008;108(6):37-42. Russian.