Vol 11, No 3 (2016)

The review offers up-to-date analysis on plant stem cell biology, meristems functioning, including plant regeneration mechanisms and tumorigenesis, as well as biotechnology methods of plant tissue culture. There were also compared the plant and animal stem cells, their probable polyphyletic origin in aspect of divergent evolution of modular and unitary organisms in both plant and animal kingdoms.

For effective transfection of the eukaryotic cells with the complexes of non-viral gene carriers and plasmids it is necessary to run a number of obstacles so as a gene construction could enter a cellular nucleus and function there successfully and long. Chemical structure of a vector has the crucial importance for the targeted complex delivery to the desired organ. At present polymers of the non-lipid origin are more and more used for gene delivery along with the lipid vectors. In the review advantages and imperfections of some classes of these less used vectors are elucidated depending on their modifications and ratio to DNA and route of the delivery. We concluded that the significant advance in a task of obtaining the effective and safe vector for the human non-viral gene delivery has not been observed yet in spite of designing more and more novel variants of the gene carriers and the new kinds of the plasmids.

Laboratory rat is one of the oldest and best studied objects of physiology and experimental medicine. However, rat pluripotent cells (embryonic and induced pluripotent stem cells) have been obtained relatively recently and remains poorly understood in terms of their transcriptome, proteome, epigenome as well as signaling cascades that maintain its self-renewal and pluripotency. In the study, the role of LIF-STAT3 cascade in maintaining self-renewal and pluripotency was investigated. It was shown that inhibition of the cascade led to cell cycle alteration, apoptotic and necrotic cell death. Additionally, RNA-seq data analysis was performed to identify transcripts level for LIF-STAT3 components. These results also suggest that LIF-STAT3 cascade plays an important role in self-renewal in rat pluripotent stem cells.

One of the promising directions of the brain tumors treatment connected using hematopoietic stem cells (HSC). The aim of this work: to study the features of the interaction between the human stem and tumor cells of different lines. The materials for the study were CD 34+-HSCs, U87 glioblastoma, lung cancer (A549), breast cancer (MCF-7). We used cell culture methods, the robotic monitoring of cellcell interactions, confocal laser microscopy, flow cytometry. It was shown that the mobility of HSCs to glioblastoma cells was higher than to cells of lung and breast cancer. The high mobility of HSCs combined with the ability to adhere to tumor cells and to share with them the contents of the cytoplasm. A direction of transport cytoplasm was depended from the tumor cell types and is a feature of the process. We believe that the described phenomenon can be one of the mechanisms of invasive growth and also indicates a high antitumor potential of HSCs.

Earlier, by an immunohistochemical method we define that after 30-day space flight in motoneurons of mice lumbar spinal cord immunoexpression of the proteins responsible for synaptic transfer of a nervous impulse and proteins of heat shock proteins decrease. In this research for an assessment of animals recovery process dynamics after space flight we studied an immunoexpression of the proteins participating in synaptic transfer of a nervous impulse (synaptophisyne, and PSD95), neurotrophic factors (a vascular endothelial factor of growth - VEGF and its receptor - Flt-1) and heat shock proteins (Hsp25 and Hsp70) in motoneurons of lumbar spinal cord of a mice after 30-day space flight on the BION-M1 biosatellite and the subsequent week readaptation to conditions of Earth gravitation. In this research by immunohistochemical method determine that after a week of animals staying in the Earth gravitation conditions the immunoexpression of synapt-ophisyne continued to decrease while the expression of PSD95, Hsp25, Hsp70 and VEGF increased in relation to the animals removed from experiment right after 30-day flight. The obtained data confirm functional plasticity of spinal cord motoneurons in the conditions of gravitation force changing. The fact, which is especially interesting, that «switches on» of neurons protective mechanisms (strengthening of heat shock proteins and neurotrophic factor expression) happens not in response to hypogravi-tation influence, but only a week after return of animals to conditions of Earth gravitation.

Bone extracellular matrix comprises a unique composite compound including the mineral and organic components. Therefore, use of biomimetic approach to the formation of tissue-engineered constructions for bone defects replacement based on composite materials containing biopolymers and calcium phosphates, as expected, can significantly improve their cyto-, biocompatibility and osteoplastic properties. The aim of the work was to study the structural features, biocompatibility and osteoplastic properties of 3D-constructions based on sodium alginate, gelatin, and two types of calcium phosphates (tricalcium phosphate and octacalcium phosphate) obtained by three-dimensional printing. The method of 3D-constructions fabrication comprised inkjet 3D-printing with hydrogel, consisted of alginate and gelatin with the addition of calcium phosphate granules, followed by freezing, freeze-drying and sterilization by y-irradiation. The structure of 3D-constructions, porosity and strength characteristics were evaluated. After the subcutaneous implantation in mice we investigated the biocompatibility of 3D-constructions during the period of up to 12 weeks. Also the osteoplastic properties of the constructions were estimated in vivo in a rat model of tibial defects. 3D printed constructions had irregular lamellar structure of sodium alginate with inclusions of spherical calcium phosphates granules. Addition of gelatin to the composite increased the porosity of constructs and significantly increased the compressive strength meanwhile practically had no effect on the ultimate strain value. In results of subcutaneous in vivo tests 3D printed constructions demonstrated perfect prolonged biocompatibility. The highest rate of biodegradation was noticed for implants containing octacalcium phosphate. All of the studied 3D-scaffolds had osteoconductive potential, more pronounced according to the number of examined histological parameters in those, made from sodium alginate, gelatin and octacalcium phosphate. The data showed the feasibility and prospect of using three-component mineral polymer composite materials based on alginate, gelatin and octacalcium phosphate as an “ink” for 3D printing of bone grafting constructions intended for implantation in bone defects.

The potential of the automated cell culture system CompacT SelecT (TAP Biosystems, UK) allows using it for the large scale production of dermal fibroblasts for cell therapy. Automated maintenance and expansion of fibroblasts requires evaluation of the proper seeding density and defining the time for the achievement of the optimal confluency of cell monolayer for further passaging. Objective - to determine the optimal parameters of dermal fibroblasts passaging for development of standardized protocol for automated cell cultivation using CompacT SelecT system. Human dermal fibroblasts were isolated from donor skin samples by enzyme dissociation. Cells proliferation rate was evaluated by the calculation of proliferation index. The dependence of proliferation rate from cell monolayer density was determined using the value of average number of divisions of single cell per day. The dependence of fibroblasts proliferation rate from seeding density (from 50 to 1.2х104cells/cm2) in various periods of cultivation (3rd, 5th, 7th and 9th day) was investigated. The inverse correlation between proliferation rate and seeding density, together with dramatic decrease of proliferation rate at fibroblasts monolayer confluency of (3.5-4.5)х104 cells/cm2, was demonstrated. The optimal seeding density was evaluated as 3х103 cells/ cm2, the most suitable density of cell monolayer for further passaging was evaluated as 3.5х104 cells/cm2. In case of high seeding density (1.2х104 cells/cm2), on day 7, when the cell monolayer density reached the critical value, the cell extinction occured, followed by renewal of cell growth. This phenomenon may be explained by the selection of cells capable of replication under conditions of high density of monolayer.

EDTA-associated pseudothrombocytopenia a laboratory phenomen associated with the widespread introduction into laboratory practice of hematology analyzers and standardized methods for blood samples preserving. High frequency pseudothrombocytopenia among healthy individuals (20%) and among patients with various pathological conditions (50%), and lack of control of the results, can cause a large amount of laboratory and diagnostic errors. Currently there is no reliable data on the clinical value pseudothrombocytopenia. Identify EDTA-pseudothrombocytopenia based on a comparison of the data automated hematological analysis with the results of microscopic examination platelets count or automated studies using alternative anticoagulants. Detection of somatic mutations in the platelet fibrinogen receptor gene (integrin gene polymorphism ITGB3-b: 1565 T>C) in a limited study - 63.2% (12 patients surveyed from 1 9) may serve as the beginning of a new direction in understanding the etiopathogenesis and clinical significance of pseudothrombocytopenia.

The features of changes in blood and bone marrow of patients with reactive (1 group, n = 8) and clonal (2 group, n = 13)eosinophilia. Revealed correlations between cellular elements, depending on the etiology, a statistically significant difference between the two independent groups of patients in terms of cellular composition of the blood and bone marrow. A common feature of both groups was high blood eosinophil counts and eosinophilic hyperplasia of the bone marrow. The absence of the correlation of some indicators of peripheral blood and hematopoietic progenitors in the bone marrow in patients 1 and 2 groups. Compared with patients with reactive eosinophilias number of correlations between indicators of hemogram and myelogram in patients with klonal eosinophilias were significantly less. Identified weak false-positive relationship between the number of neutrophils in the blood and bone marrow's neutrophils with monoclonal eosinophilias. Correlation of eosinophils and basophils blood in patients with klonal eosinophilias has been declared negative (p>0.05). On the contrary, there was a clear correlation between the two groups of red blood cells and eosinophils and their precursors. It is suggested the possible disregulatory process occurring under the action of the cytokine environment hematopoietic cells in hematologic malignancies.

Rheumatoid arthritis is a systemic autoimmune inflammatory disease of unknown etiology characterized by chronic inflammation of the synovial joints and progressive destruction of cartilage and bone. Serum and synovial fluid of RA patients revealed a wide range of autoantibodies. The highest value in the laboratory diagnosis of the disease serological tests is related to the determination of rheumatoid factor and antibodies to cyclic citrullinated peptide. In rheumatoid arthritis it is often combined with other autoimmune disorders, in particular - with autoimmune thyroiditis. Modern literature suggests that autoimmune thyroiditis and rheumatoid arthritis share common mechanisms of pathogenesis. The aim of the work was to determine the clinical and laboratory features of rheumatoid arthritis in association with autoimmune thyroiditis. We examined 146 patients with this pathology and different immunological profile. The study was conducted in three phases. In the first phase only the effect of the antibodies was determined by cyclic citrullinated peptide on the clinical and laboratory features of rheumatoid arthritis in combination with autoimmune thyroiditis. The second phase determined the effect of rheumatoid factor and antibodies to cyclic citrullinated peptide. In the third phase assessed the role of antibodies to cyclic citrullinated peptide and antibodies to citrullinated vimentin modified. The first phase showed a significant link between the immunological activity of the disease and disease activity indices, the second phase detected reliable correlation between rheumatoid factor presence in blood and age, on the third phase we found a link between immunological activity and joint destruction. Based on these data we can conclude that in patients with different immunological profile suffering from rheumatoid arthritis in combination with autoimmune thyroiditis, there were various pathogenic mechanisms of the disease.

Chronic myeloproliferative neoplasms are a group of disorders characterized by clonal disturbance of pluripotent bone marrow stem cells, leading to an excessive increase in hematopoietic cells that retain the ability to differentiate. Modern screening techniques have made changes to the current view of the epidemiology of myeloproliferative neoplasms. Recently, more and more researches are found with the median age below 50 years, and sometimes less than 40 years. This circumstance determines the need to improve therapy with the use of interferon-а. A new stage in the treatment of myeloproliferative neoplasms is the use of pegylated forms of interferon. A feature of this drug is more uniform release of interferon from copolymer combinations that results in a more effective action of the basic substance, reduced maximum weekly dose of the drug, reduce the severity of side effects, better tolerability, as well as better control of the disease progression. At the moment, even the proportion of patients receiving conventional interferon-а is very small. Therapy with pegylated interferon insufficiently developed. There are only a number of retrospective studies in recent years, indicating the usefulness of pegylated interferon to control the disease and improve the prognosis. According to the work pegylated interferon therapy is the most modern and effective approach to the treatment of chronic myeloproliferative neoplasms. We have studied the possibility of using of various forms of interferon-а for the treatment of myeloproliferative neoplasms.