[Modulation of endothelial factors activity in human endothelial cells in influenza A(H1N1)pdm09 virus infection].

INTRODUCTION: Influenza A virus infection can lead to endothelial dysfunction (ED), including apoptosis of endothelial cells and modulation of endothelial factor activities. Affected biochemical factors may include those playing important roles in vascular homeostasis. However, the effect of this pathogen on the expression pattern of key endothelial factors is still unknown.The aim of this work was to study the expression of endothelial nitric oxide synthase (eNOS) and plasminogen activator inhibitor-1 (PAI-1, serpin E1) in the EA.hy926 endothelial cells. RESEARCH OBJECTIVES: to assess expression of eNOS and PAI-1 in endothelial cells infected with influenza virus A(H1N1)pdm09, and to identify homologous fragments in structure of viral proteins and endothelial factors. MATERIAL AND METHODS: Cells were infected with influenza virus A/St. Petersburg/48/16 (H1N1)pdm09 and analyzed in dynamics in 6, 12, 18, 24, 48, and 72 hrs post infection (hpi). Detection of endothelial factors expression levels was performed by immunocytochemical method (ICC) using antibodies for eNOS and PAI-1 while quantitative assessment of expression levels was carried out by program Nis-Elements F3.2 («Nikon¼, Japan). The search for homologous sequences between viral proteins and eNOS and PAI-1 was performed by computer comparison. Sequences were analyzed as fragments 12 amino acid residues (aar) in length. RESULTS AND DISCUSSION: eNOS expression in infected cells had decreased to 7.9% by 6 hpi (control was taken as 100%) to 3.3% at 72 hpi. PAI-1 expression varied significantly over the course of the experiment: by 6 hpi it had decreased to 49.6%, and to 43.2% by 12 hpi. Later PAI-1 levels were: 116.3% (18 hpi); 18.9% (24 hpi); 23.5% (48 hpi), and 35% (72 hpi). CONCLUSION: These results indicate that influenza A infection of endothelial cells causes a significant decrease in eNOS expression, while modulating PAI-1 one. The described phenomenon can be used in the further development of directions of pathogenetic therapy of vascular complications of infection caused by this pathogen.

Generation of Reactive Oxygen Species by Human Whole Blood Cells Exposed to Iron Oxide Magnetic Nanoparticles Coated with Different Shells.

We compared dose-dependent influence of magnetic nanoparticles (MNP) coated with different shells on ROS generation during incubation with whole human blood. ROS generation was evaluated using luminol dependent chemiluminescence. MNP with polylactide shell in concentrations of 0.2-2.0 mg/ml had no effect on spontaneous chemiluminescence, while MNP with polysaccharide shell induced a dose dependent enhanced spontaneous chemiluminescence that increased during incubation (1-3 h). MNP with albumin shell in concentrations of 0.2 mg/ml gradually enhanced spontaneous chemiluminescence during incubation, but increasing the concentration neutralized this effect.

Effects of Bariatric Surgeries on the Size of Myocardial Infarction and Ghrelin Level in Rats with Experimental Decompensated Type 2 Diabetes Mellitus.

The effects of bariatric surgeries (sleeve gastrectomy and ileal transposition) on the dynamics of changes in ghrelin level were studied in rats with severe decompensated type 2 diabetes mellitus under conditions of glucose challenge as well as on the size of myocardial infarction in these animals. Diabetes was modelled by high fat diet and a single administration of streptozotocin (25 mg/kg, intraperitoneally). Both bariatric surgeries significantly decreased glucose-induced ghrelin level in the blood of rats with type 2 diabetes mellitus, which attested to an increase in the tissue sensitivity to ghrelin. Sleeve gastrectomy resulted in a decrease in the size of myocardial infarction in diabetic rats, which was calculated as the ratio of the necrosis zone to the zone of the risk of myocardial infarction. Ileal transposition had no effect on this parameter. Our data can be used as the basis for optimization of treatment approaches when using bariatric surgery in the treatment of patients with severe forms of type 2 diabetes mellitus with a high risk of cardiovascular diseases.

Multimodal Optical Diagnostics of Glycated Biological Tissues.

Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia accompanied by the disruption of carbohydrate, lipid, and proteins metabolism and development of long-term microvascular, macrovascular, and neuropathic changes. This review presents the results of spectroscopic studies on the glycation of tissues and cell proteins in organisms with naturally developing and model diabetes and in vitro glycated samples in a wide range of electromagnetic waves, from visible light to terahertz radiation. Experiments on the refractometric measurements of glycated and oxygenated hemoglobin in broad wavelength and temperature ranges using digital holographic microscopy and diffraction tomography are discussed, as well as possible application of these methods in the diabetes diagnostics. It is shown that the development and implementation of multimodal approaches based on a combination of phase diagnostics with other methods is another promising direction in the diabetes diagnostics. The possibilities of using optical clearing agents for monitoring the diffusion of substances in the glycated tissues and blood flow dynamics in the pancreas of animals with induced diabetes have also been analyzed.

Glycerol dehydration of native and diabetic animal tissues studied by THz-TDS and NMR methods.

The optical clearing method has been widely used for different spectral ranges where it provides tissue transparency. In this work, we observed the enhanced penetration of the terahertz waves inside biological samples (skin, kidney, and cornea) treated with glycerol solutions inducing changes of optical and dielectric properties. It was supported by the observed trend of free-to-bound water ratio measured by the nuclear magnetic resonance (NMR) method. The terahertz clearing efficiency was found to be less for diabetic samples than for normal ones. Results of the numerical simulation proved that pulse deformation is due to bigger penetration depth caused by the reduction of absorption and refraction at optical clearing.

[PHARMACOLOGICAL CARDIOPROTECTION DURING REPERFUSION OF ISOLATED HEART].

PURPOSE: To study the contractile function, the degree of damage and regional myocardial metabolism in the isolated rat heart model subjected to cardioplegic stop and reperfusion under the protection of levosimendan. MATERIALS AND METHODS: The study was performed on isolated rat hearts Wistar (group using "Custodiol" vs group using "Custodiol" + "Levosimendan". We assessed the extent of myocardial damage (in terms of markers of myocardial necrosis), the contractile function of the myocardium (coronary flow, heart rate, left ventricular pressure), the dynamics of redox processes during reperfusion with a parallel study of histology of the myocardium. RESULTS: We found a presence of cardioprotective effect of levosimendan in respect of the isolated heart in the reperfusion period of cardioplegic ischemia. The effect related to reducing the emission of reperfusion enzyme markers of myocardial damage, reducing the severity of pathological changes in the myocardium and reducing the intensity of free radical reactions in the myocardium. CONCLUSIONS: Cardioprotection with levosimendan reduces the severity of free radical attack the isolated heart, reduces the severity of damage to cardiomyocytes and preserves the contractile activity of the myocardium during reperfusion due to the effect of postconditioning.