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Synthetic materials used in regenerative medicine, upon implantation, induce the development of an inflammatory reaction necessary for the effective regeneration of damaged bone tissue. Implant contact with tissues is accompanied by the deposition of blood proteins and interstitial fluid on its surface, contributing to the activation of the complement system, components of innate immunity, initiating coagulation hemostasis, leading to the formation of a fibrin clot. An extracellular matrix based on fibrin, collagen and elastin forms on the implant’s surface, which provides the basis for the formation of tissue structure through the adhesion of stem cells to the forming bone callus before the formation of bone regenerate. To prevent the development of postoperative pathological conditions caused by hypercoagulable syndrome, therapeutic strategies are used to use anticoagulants (heparin, warfarin). However, their use limits the normal formation of a fibrin clot in vivo. This can slow down the migration of mesenchymal stem cells (MSC) and disrupt the formation of callus, inhibiting the processes of osseointegration of the implant and bone healing. The study’s goal was to study the effect of heparin in a gradient of low and high concentrations on the migration activity and stem capacity of human MSCs underin vitrocultivation conditions. According to the results of flow cytometry, it was revealed that high concentrations of heparin (130, 260 IU/ml) in a 2D cultivation model contribute to an increase in the number of cells expressing surface markers CD73 and CD90, which indicates that MSCs retain high clonogenic potential. A 3D model ofin vitrocultivation with the addition of heparin and osteosubstituting implants bearing a CF coating with a roughness index of Ra = 2.6-4.9 μm contributed to preserving the “stemness” character of MSCs through the expression of surface markers CD73 and CD90. According to the results obtained using the xCELLigence system, heparin at a later time (from 20-40 hours) increases the invasion of MSCs through micropores that simulate the state of the blood vessel walls. However, in the presence of HAP nanoparticles that mimic the remodeling processes of the mineral bone matrix and/or resorption of bone cement, the effect of heparin was less pronounced. The results can be used in the field of regenerative medicine associated with the introduction of MSCs. The data can serve as a prerequisite for developing new therapeutic strategies for surgical patients with a high risk of postoperative thrombosis after osteosynthesis. more...

In obese patients, the relationship between the content of chemerin in blood plasma and the expression of genes TFAM, Drp1, MFN2, SOD, BAX, responsible for quality control of mitochondria, in insulin-dependent tissues (adipose tissue, liver) was revealed. The tissue-specific features of gene expression (TFAM, Drp1, MFN2, SOD, BAX), the number of mtDNA copies in the studied depots in obese patients were established. It has been proven that a change (decrease) in the number of mtDNA copies in insulin-dependent tissues can have a protective effect on mitochondria under conditions of increased oxidative stress. It was found that in patients without type 2 diabetes, an increase in chemerin production promotes the activation of the antioxidant system in the visceral adipose tissue but not in the liver. On the contrary, all obese patients with type 2 diabetes showed a decrease (compared with patients without type 2 diabetes) in the plasma level of chemerin. Thus, the low content of chemerin in the blood plasma in patients with type 2 diabetes mediates the formation of mitochondrial dysfunction in insulin-dependent tissues (adipose tissue, liver). more...

Steatosis in the liver in obesity increases the work of mitochondria to utilize excess lipids. An overload of β-oxidation of fatty acids, the tricarboxylic acid cycle, and oxidative phosphorylation leads to a decrease in ATP and an increase in the formation of reactive oxygen species. Normally, mitochondria can efficiently remove elevated levels of reactive oxygen species using the cell's antioxidant system and metabolic adaptation to altered conditions. This study aimed to investigate the role of hepatic SOD expression in the pathogenesis of NAFLD in obesity. It was found that the level of SOD1 expression in the liver in obese patients with and without type 2 diabetes with a BMI > 40 kg/m2 was lower than in healthy donors. The copy number of mitochondrial DNA (mtDNA) in the liver in all obese patients was more than two times lower than in the control group. In the liver of obese patients without type 2 diabetes, the SOD1 protein level and the mtDNA copy number were interrelated and negatively correlated with the area of fatty inclusions. Thus, in obese patients, a decrease in antioxidant defense in the liver leads to the vulnerability of mitochondria, which, in turn, contributes to the progression of steatosis and insulin resistance. more...

The development of “biohybrid” drug delivery systems (DDS) based on mesenchymal stem/stromal cells (MSCs) is an important focus of current biotechnology research, particularly in the areas of oncotheranostics, regenerative medicine, and tissue bioengineering. However, the behavior of MSCs at sites of inflammation and tumor growth is relevant to potential tumor transformation, immunosuppression, the inhibition or stimulation of tumor growth, metastasis, and angiogenesis. Therefore, the concept was formulated to control the lifespan of MSCs for a specific time sufficient for drug delivery to the target tissue by varying the number of internalized microcontainers. The current study addressed the time-dependent in vitro assessment of the viability, migration, and division of human adipose-derived MSCs (hAMSCs) as a function of the dose of internalized polyelectrolyte microcapsules prepared using a layer-by-layer technique. Polystyrene sulfonate (PSS)—poly(allylamine hydrochloride) (PAH)-coated spherical micrometer-sized (diameter ~2–3 µm) vaterite (CaCO3) microcapsules (PAH-PSS)6 with the capping PSS layer were prepared after dissolution of the CaCO3 core template. The Cell-IQ phase contrast imaging results showed that hAMSCs internalized all (PAH-PSS)6 microcapsules saturating the intercellular medium (5–90 particles per cell). A strong (r > 0.7) linear dose-dependent and time-dependent (up to 8 days) regression was observed between the in vitro decrease in cell viability and the number of internalized microvesicles. The approximate time-to-complete-death of hAMSCs at different concentrations of microcapsules in culture was 428 h (1:5 ratio), 339 h (1:10), 252 h (1:20), 247 h (1:45), and 170 h (1:90 ratio). By varying the number of microcontainers loaded into the cells (from 1:10 to 1:90), a dose-dependent exponential decrease in both the movement rate and division rate of hAMSCs was observed. A real-time cell analysis (RTCA) of the effect of (PAH-PSS)6 microcapsules (from 1:5 to 1:20) on hAMSCs also showed a dose- and time-dependent decrease in cell longevity after a 50h study at ratios of 1:10 and 1:20. The incorporation of microcapsules (1:5, 1:20, and 1:45) resulted in a dose-dependent increase in 24–48 h secretion of GRO-α (CXCL1), MIF, and SDF-1α (CXCL12) chemokines in hAMSC culture. In turn, the normalization or inhibition of chemokine secretion occurred after 72 h, except for MIF levels below 5–20 microcapsules, which were internalized by MSCs. Thus, the proposed concept of controlling the lifespan of MSC-based DDS using a dose of internalized PAH-PSS microcapsules could be useful for biomedical applications. (PAH-PSS)6 microcapsule ratios of 1:5 and 1:10 have little effect on the lifespan of hAMSCs for a long time (up to 14–18 days), which can be recommended for regenerative therapy and tissue bioengineering associated with low oncological risk. The microcapsule ratios of 1:20 and 1:45 did not significantly restrict the migratory activity of hAMSCs-based DDS during the time interval required for tissue delivery (up to 4–5 days), followed by cell death after 10 days. Therefore, such doses of microcapsules can be used for hAMSC-based DDS in oncotheranostics. more...

Background: Despite the great interest and numerous studies, there is currently no unified standard describing the sequential manipulation with cells to obtain exosomes for clinical use.The use of exosomes has become an attractive alternative to cell therapy, since the flexible nature of these biological vesicles allows scientists to manipulate their composition to produce the desired exosomes carrying specific drugs, RNA and proteins. This study aimed to analyse scientific literature on the changes in the functional characteristics of exosomes, depending on the method of manipulation, potentially contributing to the development of negative effects in the treatment of diseases of inflammatory genesis. Results: The choice of isolation method affects the expressed sets of protein markers, nucleic acids and receptors on microparticles. Various surface receptors present on the exosome membrane can be engineered to target lesions. Exosomes from healthy patients help to reduce inflammation, normalize intercellular communication and have anti-fibrotic, antioxidant, and cytoprotective effects. Exosomes can change the microenvironment, but the microenvironment can also change the composition of exosomes. Conclusion: Exosomes obtained from sick patients carry markers characteristic of the corresponding disease. Such exosomes can have pro-inflammatory, pro-fibrotic, cytotoxic, and oncogenic properties, and disrupt cellular cooperation. Until now, questions regarding the dose, reactions to repeated administration, and dosage regimes have not been completely resolved. more...

T-lymphoblast-like human leukemia cells of the Jurkat line (Jurkat T cells) form polyploid forms with a high DNA content in suspension culture. Due to pronounced genetic instability, this contributes to further transformation and development of clonal diversity (polyclonal) of the cell line. Little information is available on the adherent subpopulation of Jurkat T cells. In this work, we analyzed the content of nucleic acids in suspension (DNA) and adhesive (DNA, RNA) subpopulations of Jurkat T cells using flow cytometry and propidium iodide dye, as well as confocal laser microscopy and acridine orange dye. The morphology and mobility of large (with a diameter of more than 15 μm) Jurkat T cells adhering to plastic were studied using Cell-IQ phase-contrast microscopy in real-time. According to the intensity of fluorescence in the conditionally green wavelength range (300–530 nm: from UV to green) and conditionally red (565–800 nm: from red to far-red), three subpopulations of adherent Jurkat T cells were identified: with high, medium, and low nucleic acid content. Thus, Jurkat-T cells adhering to the plastic surface of the plates retain a pronounced heterogeneity in the DNA content characteristic of the suspension fraction, which suggests a difference in the morphofunctional properties (polyclonicity) of this subpopulation of cell culture. With a sharp increase in the total cell mass, the proportion of large (giant, 15–50 μm or more) cells attached to the plastic remained constant for 21 days of cultivation and amounted to 1% of the adhesive fraction. It was found that large Jurkat T cells (with a median diameter of 31 µm) moved along the plastic at a linear (median) speed of 38 µm/h. The polynuclear Jurkat T cells on plastic are morphologically identified, with a linear increase being revealed in the mobility of adherent cells with an increase in their diameter (regression coefficient r = 0.33, p < 0.02, n = 52). Possible cellular and molecular mechanisms of an increased number of DNA copies in some adhering Jurkat T cells are discussed. It is assumed that the discovered new property (locomotor activity) can provide polyploid (multinucleated) adhering Jurkat T cells with a significant advantage—directed migration (chemotaxis) in a growing cell population under conditions of nutrient deficiency due to a change in the nutrient medium after 3–4 days of cultivation. more...

Experimental titanium implants with a rough calcium phosphate (CР) coating with bio-inspired properties have been developed. The role of roughness (in the range Ra = 2.4 - 4.6 μm) of the surface CP simulating the physicochemical features of the mineral substance of regenerating bone tissue to change the growth and morphofunctional activity of mesenchymal stem cells (MSCs) was evaluated. Titanium substrates with a microrelief CP coating bearing artificial niches for stem cells was shown in vitro to have an epigenomic effect on MSCs, that contributes to their differentiation and maturation in bone cells. The results show the promise of developing and introducing a new class of medical devices with bio-inspired surfaces into clinical practice for traumatology and orthopedics. more...

The paper reports a study of the morphofunctional activity of T-lymphocytes in response to the in vitro contact with calcium phosphate (CP) coating in the presence of the particles bearing antibodies against CD2, CD3, and CD28 antigens. VT1-0 titanium plates (10 × 10 × 1 mm3) with a double-sided microarc rough (Ra = 2–5 µm) CP coating were used as the model samples of the mineral matrix of the bone tissue. Magnetic particles (MACSiBeadTM T-cell Activation/Expansion Kit human) bearing antibodies to CD2, CD3, and CD28 antigens were used as a T-cell activator (TCA) simulating the signals produced by antigen-presenting cells (APCs). Mononuclear cells (MNCs) isolated from human blood (98.8% of CD45CD3+ cells) were cultured in the presence of samples with a CP coating and/or TCA (2 × 106 particles in 1.5 mL of nutritive medium in the proportion of 2 : 1 to cells) for 2 and 14 days. The CP coating and TCA triggered MNC culture adaptation in a synergic way via the mechanisms of hyperactivation and subsequent death of T-lymphocytes. Immune selection occurred through the accumulation of the naive CD45RA+/RO+ T-lymphocytes and memory T-cells with the simultaneous depletion of the CD4+ and CD8+ T-cell pool. The shift in the T-lymphocyte populations was observed together with the increase (after 48 h of culturing) in the cell secretion activity with its subsequent decrease by the 14th day of observation. CP coating sustained (compared with the cell culture grown on plastic) the secretion ability of Th1 (IL-12, TNFα, and IFNγ) and Th2 (IL-4, IL-6, IL-10, and IL-13) lymphocytes. At the same time, the prolonged TCA signal after the 48-h activation caused the depletion of T-cell secretion. The suggestion that the observed in vitro effects may play role in the switching of signaling between T-lymphocytes, APCs, and CP materials at the cell–foreign body interface, which may result in a change in the inflammation phase, development of immune tolerance, successful osseointegration of the implant, or bone tissue remodeling impairment, is discussed. more...

The role of γc cytokines (IL-2, IL-7, and IL-15) in the regulation of apoptotic death of memory T cells under cultivation conditions in vitro was studied using the method of cytofluorimetry. It was detected that the effect of IL-2 in combination with TCR activator is aimed at increasing the number of mature CD45RO+CD95+ T cells with a simultaneous decrease in the number of viable T cells, which gives reason to assume the proapoptotic effect of IL-2 under restimulation conditions. On the contrary, the effect of IL-7 and IL-15 in the activation model in vitro leads to an increase in the number of living T cells in СD45RO cultures (as compared with the samples, in which only the activator was added) against the background of an increase in the number of CD8+CD95+ T cells with a central memory phenotype (TCM). Changes induced by IL-15 are aimed an decreasing the content of CD4+CD95+ T cells in CD45RО+ cultures of the immune memory T cells. Thus, a dose-dependent positive effect of IL-2 on the clonal expansion and negative effect on the viability of T cells was demonstrated. While IL-7 and IL-15, on the contrary, preserve the viability of T cells, participating in the generation of a pool of long-lived memory T cells. more...

Aim. Search for the association of polymorphisms C(-15161)T (rs10902088) and T(-12150) C(rs10794288) of the MUC2 gene and С(-344)Т (rs1799998) of the CYP11B2 gene with the increased risk of genital endometriosis development in the Slavic population. Materials and methods. The study included 85 female patients (from 19 to 44 years old) with a diagnosis of genital endometriosis and 79 healthy women (from 20 to 42 years old). Genotyping was performed by real-time PCR using a Light Cycler 480 Instrument II amplifier (Roche, Switzerland). Polymorphism determination kits, MUC2 C(-15161)T (rs10902088), MUC2 T(-12150)C (rs10794288), CYP11B2 С(-344)Т (rs1799998) (CJSC Syntol). Results. It was found that the CC genotype of the T(-12150)C (rs10794288) C polymorphism of the MUC2 gene and the C(-344)T (rs1799998) CC polymorphism genotype of CYP11B2 gene are protective against endometriosis, and the TT genotype of C(-344)T polymorphism ( rs1799998) of the CYP11B2 gene contributes to the development of this disease. Conclusion. The search for molecular genetic aspects in the development of genital endometriosis is necessary for early diagnosis and prognosis of the disease. more...

This paper focuses mainly on the in vitro study of a five-week biodegradation of a-C:H:SiO

The major challenges in Osteosarcoma (OS) therapy are its heterogeneity and drug resistance. The development of new therapeutic approaches to overcome the major growth mechanisms of OS is urgently needed. The search for specific molecular targets and promising innovative approaches in OS therapy, including drug delivery methods, is an urgent problem. Modern regenerative medicine focuses on harnessing the potential of mesenchymal stem cells (MSCs) because they have low immunogenicity. MSCs are important cells that have received considerable attention in cancer research. Currently, new cell-based methods for using MSCs in medicine are being actively investigated and tested, especially as carriers for chemotherapeutics, nanoparticles, and photosensitizers. However, despite the inexhaustible regenerative potential and known anticancer properties of MSCs, they may trigger the development and progression of bone tumors. A better understanding of the complex cellular and molecular mechanisms of OS pathogenesis is essential to identify novel molecular effectors involved in oncogenesis. The current review focuses on signaling pathways and miRNAs involved in the development of OS and describes the role of MSCs in oncogenesis and their potential for antitumor cell-based therapy. more...

Obesity and osteoporosis are global health problems characterized by high rates of prevalence and mortality due to complications. As people with visceral obesity age, the adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) increases, and adipocytes become the predominant stromal cells in the bone marrow microenvironment, which hinders the physiological regeneration and mineralization of bone tissue. Primary and secondary osteoporosis remain severe progressive diseases. Both osteoporosis and obesity are associated with microRNAs (miRNAs) that induce adipogenesis and osteoresorption. This review presents analyses of the roles and clinical potential of miRNAs in the epigenetic control of BMSC differentiation and the formation and function of osteoclasts in osteoporosis with and without obesity. Understanding the fine-tuned regulation of the expression of genes critical for the balance of osteogenesis/osteolysis processes may provide hope for the development of effective and safe osteoporosis therapies in the future. more...

Novel polyfluorene polymer brushes with polymethacrylic acid side chains were obtained by atom transfer radical polymerization (ATRP) and activator generated by electron transfer (AGET) ATRP of tert-butyl methacrylate on polyfluorene multifunctional macroinitiator, followed by protonolysis of the tert-butyl groups of the side chains. Kinetics of polymerization and molecular weights were fully characterized. These polymer brushes luminesce in the blue region of the spectrum with high quantum yields (0.64–0.77). It was shown that the luminescence intensity of polymer brushes is higher than the luminescence intensity of the macroinitiator (0.61). Moreover, due to their amphiphilic nature, they can form unimolecular micelles when an alcohol solution of the polymer brush is injected into water. These properties can potentially be used in drug delivery and bioimaging. more...

The article covers the main biotechnology tools - cloning, genetic engineering (recombinant technology) technologies, genome editing, producing transgenic animals, and examples of each tool's practical applications. For each method, the characteristics that determine the scope of its applications are described. more...

Subclinical inflammation in morbid obesity is associated with immune activation and the development of concomitant diseases. Impaired immune homeostasis and immune cell dysregulation in adipose tissue are associated with phenotypic and functional changes in the pool of T lymphocytes and the development of chronic hypovitaminosis D. Low vitamin D levels in obesity lead to the activation, proliferation and production of pro-inflammatory mediators by T cells. Hypovitaminosis D is the cause of a decrease in the functional potential of regulatory and anti-inflammatory lymphocytes and the maintenance of the inflammatory response. The exact molecular genetic mechanisms of the effect of vitamin D on T lymphocytes have not been fully elucidated. Therefore, uncovering the functional role of T cells and their relationship to vitamin D homeostasis in the context of obesity development may contribute to the development of new pathogenetic methods for clinical prediction of the risk of metabolic, oncologic, autoimmune and infectious complications. The review presents the molecular genetic mechanisms of the effect of vitamin D on adipose tissue resident T lymphocytes and the characteristics of vitamin D receptor expression, and analyzes the phenotypic and functional characteristics of potentially pathogenic T lymphocytes in relation to the development of obesity and its associated complications. more...

The review discusses the complex, ambiguous and individual effects of heparin and its derivatives on the bone and circulatory systems, in dependence of the dosage, the state of the cells and tissues of recipients. General data on the anticoagulant activity of heparin and its derivatives are presented; aspects of the effect of heparin on mesenchymal cells and tissues and its role in angiogenesis are considered in details. Particular attention is paid to the ability of heparin to bind osteogenic and angiogenic biomolecules: thus us especially important for the development of systems for their delivery and sustained controlled release. A schematic representation of the positive and side effects of heparin as a delivery system for biomolecules in tissue engineering is proposed.V obzore obsuzhdaiutsia slozhnye, neodnoznachnye i individual'nye éffekty geparina i ego proizvodnykh na kostnuiu i krovenosnuiu sistemy v zavisimosti ot dozirovki, sostoianiia kletok i tkaneĭ retsipienta. Predstavleny obshchie dannye o protivosvertyvaiushcheĭ aktivnosti geparina i ego proizvodnykh; detal'no rassmotreny aspekty vliianiia geparina na mezenkhimnye kletki i tkani i ego rol' v angiogeneze; obsuzhdaetsia sposobnost' geparina sviazyvat' osteogennye i angiogennye biomolekuly v prilozhenii k razrabotke sistem ikh dostavki i dlitel'nogo kontroliruemogo vysvobozhdeniia. Predlozheno skhematicheskoe predstavlenie o pozitivnykh i pobochnykh éffektakh geparina kak sistemy dostavki biomolekul pri inzhenerii tkaneĭ. more...

BACKGROUND: Vitamin D binding protein is a main vitamin D carrier in serum. It also has an impact on macrophagial function. Role of vitamin D and macrophages in the pathogenesis of atherosclerosis is scientifically proven but there is lack of data on vitamin D binding protein in this regard. AIMS: To evaluate the vitamin D binding protein polymorphism in patients with acute coronary syndrome without diabetes mellitus, autoimmune diseases and malignant tumors. Determine correlation, if there is, between vitamin D binding protein allele and features of acute coronary syndrome among this patient group. MATERIALS AND METHODS: It is a cross-sectional observational study. Study subjects are patients with acute coronary syndrome. Exclusion criteria are the presence of diabetes mellitus, autoimmune diseases and malignant tumors. In all participants were evaluated: predisposing factors for heart diseases, CBC, biochemical blood test, troponin, coronarography, echocardiography. The study lasted for 5 months from November 2017 until March 2018. Primary end point – assessment of vitamin D binding protein polymorphysm in this group of patients with acute coronary syndrome by means of vitamin D binding protein gene sequencing. 50 patients were enrolled into this study who were urgently admitted to hospital and diagnosed with acute coronary syndrome. Among them – 36 males and 14 females. Mean age was 60 (55;66) years. All participants were sequenced for single nucleotide polymorphysm in VDBP p.T436K (rs4588) and P.432E (rs7041). RESULTS: Gene polymorphysms of interest were found in 43 patients among 50 enrolled. Haplotype Gc1s/2 (rs7041G-rs4588A) was found in 7 (14%) patients, Gc2 (rs7041T-rs4588A) — in 9 (18%) patients, Gc1s (rs7041G-rs4588C) – in 20 (40%) patients, Gc1f (rs7041T-rs4588C) in 14 (28%). Coronarography showed that coronary artery occlusions obstructing more than 50% of vessel lumen was found in 16 patients; obstruction greater than 90% was seen in 8 patients; total occlusion – in 4 patients. CONCLUSIONS: In patient group with acute coronary syndrome prevalence of vitamin D binding protein gene polymorphysm was high – in 86% of participants. The features of Gc2 haplotype were higher frequency of recurrent myocardial infarction and total coronary artery occlusion, as well as tendency to decreased serum vitamin D3 (25(OH)D) levels. more...

In modern medicine, personalized approaches to the diagnosis, prevention and treatment of diseases are becoming increasingly important. The accumulated information on genetic mutations and their mapping led to the development of prognostic genetic tests. Created programs for clustering and analysis of the information received have served the development of pharmacogenetics and gene therapy. This has increased the effectiveness and safety of therapeutic approaches and increased average life expectancy. At the same time, the Russian Federation (RF) and the world community are actively discussing the protection of human rights, the inviolability of the person, storage, use of his personal data, including genetic information from unauthorized access. The purpose of this review was to generalize and detail information on legislative norms and legal regulation of the use of genetic information in the territory of the Russian Federation and in other countries of the world. more...

Background: Gastric and colon tumors are often associated with eosinophilic infiltration of tumor tissue, the significance of which is still not entirely clear. The recruitment of eosinophils into the tissues can be in part regulated by galectins ― galactose-binding proteins which are expressed by a variety of tissues and are capable of exerting a broad range of effects. Aims: To evaluate the expression of galectin-1 and galectin-3 in tumor tissue, and gal-3 gene mRNA expression in blood eosinophils in patients with gastric and colon cancer with or without tissue eosinophilia. Materials and methods: The study included a total of 107 patients (84 males and 23 females, average age 60,9 6,8) with verified gastric cancer (52 persons) and colon cancer (55 persons), who underwent treatment or were registered at the dispensary at the regional medical institution Tomsk Regional Oncology Center (Tomsk, Russia). The control group consisted of 15 men and 11 women of comparable age. The materials of the research included samples of gastric and colon tumors obtained during surgery, and eosinophilic granulocytes isolated from whole blood by immunomagnetic separation. Galectin-1 and galectin-3 expression in tumor tissue was evaluated by immunohistochemistry. The expression of gal-3 gene mRNA in eosinophils was determined by real-time reverse transcription polymerase chain reaction. Statistical analysis of the results was carried out using the non-parametric Mann-Whitney U test for independent samples with Benjamini-Hochberg procedure for multiple comparisons, and the Chi-square Pearson criterion with Yates correction. Results: In patients with gastric cancer and colon cancer, regardless of the presence of tissue eosinophilia, low expression of galectin-3 in the tumor tissue and high expression of gal-3 gene mRNA in peripheral blood eosinophils were found. Gastric and colon cancer patients with eosinophilic infiltration of tumor tissue were characterized by low expression of galectin-1 within tumor cells (in 64.0% cases, 2 = 4.890, р = 0.029; and in 73.9% cases, 2 = 5.981, p = 0.031 respectively). There was a statistically significant connection between the level of galectin-1 expression by tumor cells and the presence of tissue eosinophilia both in gastric ( = 0.307) and colon cancer ( = 0.330). Conclusion: Low expression of galectin 1 and 3 by tumor cells in gastric and colon cancer with tissue eosinophilia indicates the lack of a significant effect of these proteins on the process of recruiting eosinophilic granulocytes into tumor tissue. Increased expression of galectin-3 in blood eosinophils in gastric and colon cancer is not associated with the presence of eosinophilic infiltration of tumor tissue. more...

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. more...

Using a multiplex kit the secretion of a number of cytokines, chemokines, and growth factors has been investigated in vitro in a culture of human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs) under conditions of their osteogenic differentiation caused by 14-day contact with a calcium phosphate (CP) surface of different roughness. Bilateral X-ray amorphous CP coatings were prepared on the samples of commercially pure titanium in the anodal regime using a microarc method. The electrolyte consisted of aqueous orthophosphoric acid (20 wt %), calcium carbonate (9 wt %), and synthetic hydroxyapatite nanopowder (6 wt %, particle diameter of 10–30 nm with single agglomerates up to 100 nm). hAMMSCs isolated from lipoaspirate were co-cultured after 4 passages with the CP-coated samples at a final concentration of 1.5 × 105 viable karyocytes per 1.5 mL of standard nutrition medium (without osteogenic stimulators) for 14 days (determination of the [CD45,34,14,20], CD73, CD90, and CD105 cell immunophenotype; analysis of secretory activity) and 21 days (alizarin red S cell culture staining) with medium replacement every 3–4 days. Under conditions of in vitro contact with rough CP coating hAMMSCs differentiated into osteoblasts synthesizing the mineralized bone matrix; this was accompanied by a 2−3-fold increase in the proportion of [CD45,34,14,20]+ hemopoietic cells. The following humoral factors of hemopoietic niches acted as the signal molecules escalating in vitro the hemopoietic base in 14 days of differentiating three-dimensional culture of hAMMSCs: leukemia inhibitory factor (LIF) and stem cell factor (SCF) cytokines in the case of the mean index of CP roughness Ra = 2.4–2.6 µm or stromal derived factor-1 (SDF-1α, CXCL12 chemokine) in the case of Ra = 3.1–4.4 µm. more...

We studied the role of native α-fetoprotein preparation in the regulation of proliferation and functional activity of naive T cells and immune memory T cells in vitro. The study was carried out on separated fractions of naive T cells (CD45RA+) and immune memory T cells (CD45R0+) incubated with α-fetoprotein under conditions of TCR activation. At the level of naive T cells, α-fetoprotein in a concentration of 100 U/ml reduced the expression of CD28, but increased the expression of CD25, while at the level of immune memory T cells α-fetoprotein (50 and 100 U/ml) only suppressed the expression of CD25. No effects of α-fetoprotein on the proliferative status of the studied lymphocyte subpopulations and on the expression of CD71 (proliferation marker) by these cells were detected. Addition of α-fetoprotein in a concentration of 100 U/ml increased the level of IL-2 in naive T cell culture supernatants, while production of IL-2 by memory T cells remained unchanged. These data demonstrated the priority aspects of regulation of the functional activities of naive T cells and immune memory T cells. more...

Zn-, Cu- or Ag-incorporated calcium phosphate coatings were deposited on pure titanium and Ti-40 wt% Nb substrates by the micro-arc oxidation method. Microstructure, morphology, phase composition, physico-chemical and antibacterial properties of the formed coatings and their behavior in synthetic biological media have been studied. Due to the different compositions of the electrolytes and different substrates the MAO process took place at various electro-physical parameters such as current densities and voltages. As a result, the coatings had different properties, phase composition and were characterized by a different behavior in synthetic biological fluids. When using the acidic electrolyte, the applied voltage varied in the range of 200–300 V. In the case of an alkaline electrolyte, the voltage range was 350–450 V. The current density of the MAO process in acidic electrolyte was ten times higher than that in alkaline electrolyte. Therefore, the thickness, roughness and porosity of the Zn-, Cu-incorporated calcium phosphate coatings deposited in acidic electrolyte were the highest. The Zn-, Cu-incorporated coatings showed weight gain in the biological fluids due to intensive dissolution and consequent sedimentation of the calcium phosphates on the coating surface. Ag-incorporated coatings dissolved slowly in the biological fluid. The calcium phosphate coatings containing trace elements Zn, Cu or Ag demonstrated antimicrobial activity against Staphylococcus aureus 209P and in general, did not show cytotoxic effect on myelocariocytes. more...

Currently, one of the approaches to correct metabolic disorders in the type 2 diabetes mellitus (DM2) with obesity are bariatric surgery (BS), including sleeve gastrectomy (SG), gastric bypass (GB) and ileal transposition (IT). However, their effectiveness and impact on the hypothalamic signaling and hormonal status in severe forms of DM2 without obesity remain little studied. The aim of the work was to study the effect of IT, SG and GB on the insulin, leptin, ghrelin and glucagon-like peptide-1 (GLP-1) levels in the blood and on the expression of the genes encoding the main components of the hypothalamic signaling systems in rats with decompensated form of DM2, which was induced by a high-fat diet (3 months) and a single low dose of streptozotocin (25 mg/kg, 2 months after the start of the diet). In diabetic rats, a significantly expressed hyperglycemia, an impaired glucose tolerance, a decrease in glucose-stimulated GLP-1 level, a slight decrease in the insulin and leptin levels and an slight increase in ghrelin level were detected. In the hypothalamus, the expression of the genes encoding GLP-1 receptor, orexigenic agouti-related peptide (AgRP), as well as phosphotyrosine phosphatase 1B and SOCS3, the negative regulators of the leptin and insulin pathways was increased. In diabetic rats, the IT reduced the glucose levels 120 minutes after glucose load, increased the basal and glucose-stimulated GLP-1 levels, normalized the gene expression for phosphotyrosine phosphatase 1B, SOCS3, AgRP and GLP-1 receptor, which indicates the restoration of the hypothalamic signaling responsible for the control of energy metabolism and insulin sensitivity. In the case of SG and GB, an improvement in the glucose tolerance was found, and in the case of SG, an increase in the basal and glucose-stimulated GLP-1 levels was shown. However, no significant effect on the expression of the hypothalamic genes in SG and GB was found. Thus, IT is the most effective of all studied BS in the treatment of severe forms of DM2 without obesity. more...

We studied the effects of methylprednisolone (MP) on the late stages of differentiation of activated memory T cells in healthy donors and in patients with the diagnosis of rheumatoid arthritis (RA) in vitro. It was found that MP exerts a suppressing effect on the number of CD4+ T cells expressing the activation (CD25) and co-stimulation (CD28) molecules in CD3+CD45RO+ cultures of healthy donors and patients with rheumatoid arthritis (RA). Assessment of the level of expression of the U2af1l4 and Gfi1 genes controlling the alternative splicing of the Ptprc gene demonstrated unidirectional but quantitatively different changes in health and in RA. Association between the level of expression of the U2af1l4 and Gfi1 genes and the number of CD3+CD4+CD45RO–CD28– cells in the populations of TCR-activated memory T cells in RA patients treated with MP confirms the hypothesis that glucocorticoids can stimulate the formation of terminally differentiated lymphocytes (CD4+CD28–CD45RO–, TEMRA), which are supposed to play the key role in the pathogenesis of RA. more...