Introduction. Multiparametric comparative analysis of clinical and molecular genetic biomarkers of malignant tumors has strong diagnostic and prognostic potentials and is a prerequisite for the development of personalized medicine. This approach makes it possible not only to simultaneously detect the expression of several tumor biomarkers, but also to obtain data on their spatial distribution in tissues examined, as well as to estimate the mutual location of tumor cells and tumor microenvironment expressing specific biomarkers. Thus, multiparametric immunohistochemical analysis (IHCA), which allows not only confirming the specific disease, but also carrying out 3D imaging of biopsy specimens and analyzing the spatial organization of tumor tissue, as well as the expression rates of biomarkers at the level of individual cells, opens wide prospects in the diagnosis and treatment of cancer.

Aim. Systematizing data on the potential of multiparametric IHCA for cancer diagnosis and development of the personalized approach to cancer therapy.

Results. Multiparametric IHCA allows estimating the heterogeneity of the tumor at the level of molecular subtypes, as well as the heterogeneity of the tumor microenvironment. These data make it possible to predict tumor development, determine its metastatic potential, and select an effective strategy for individual therapy.

Conclusion. This review analyzes the use of multiparametric IHCA for the detection of malignant tumors and shows its high potential for the differentiation of tumors and the study of tumor microenvironment. This ensures effective selection of the therapeutic strategy and accurate assessment of the response to therapy.

Introduction. Early sensitive and highly specific diagnosis is crucial for successful cancer therapy. The use of fluorescent hydrogels (FHG) makes it possible to develop versatile biosensors due to the increased binding capacity of biological capture and reporter molecules, sensitive fluorescence detection, and the flexibility of combining their structural and functional elements.

Aim. Analyzing the principles of designing biosensors based on FHG for the detection of cancer markers and the methodological approaches to their development, as well as summarizing and systematizing the data on the principles of detection and target signal generation used in these sensors.

Results. FHG represent 3D sensing platforms, i. e., structures that combine the reporter fluorescence function with biological capture molecules, allowing the unique optical properties of fluorescent nanocrystals at the macro level to be preserved. The porous structure of hydrogels increases the active surface area of biosensors for 3D immobilization of fluorescent labels and biological capture molecules, while preserving the structure of these molecules, which ensures specific binding of the detected molecules of the sample. This ensures a higher sensitivity compared with the traditional methods of immunoenzymatic and immunochromatographic analyses. Not only the traditionally used antibodies, but also enzymes and glycoproteins, aptamers and oligonucleotides, as well as polymers obtained by molecular imprinting, can serve as biological capture molecules, which extends the range of specifically detectable analytes.

Conclusion. The review presents examples of biosensors based on FHG intended for the detection of cancer markers and describes approaches to the preparation of FHG and immobilization of biological capture molecules, as well as principles of generation of the detected optical signal. The main advantages of fluorescent hydrogel biosensors over the classical tests used for quick diagnosis of cancer are shown.

Diagnosis of secondary myeloid neoplasms (therapy-related myeloid neoplasms) associated with therapy of solid tumors, in most cases, is not associated with significant difficulties. The problem is the diagnosis of secondary myelodysplastic syndromes after the treatment of acute myeloid leukaemias. The complexity of early diagnosis of secondary myelodysplastic syndromes is due to the differentiation of this nosology and the early recurrence of previous acute myeloid leukemia and, as a result, the difficulties of prognosis and risk stratification for therapeutic management. The relevance of this problem is explained by the rare case reports. Making the diagnosis of secondary myelodysplastic syndrome, in our opinion, can be based on the absence of a connection of cancer cell clone with the first (previous) disease in a molecular study. In this publication, we describe the first domestic case report of myelodysplastic syndrome diagnosed after chemotherapy for acute myeloid leukemia, based on differences in cytomorphology, immunophenotyping and molecular research. we interpreted the prognosis as favorable and prescribed appropriate treatment.

Background. Interleukin-10 (IL-10) is a pleiotropic cytokine with immunomodulatory properties and may inhibit tumor development and progression or stimulate tumor growth.

Aim. Analysis of the changes of the IL-10 mRNA level in the peripheral blood (PB) of patients with prostate cancer (PC) and benign prostatic hyperplasia (BPH) in comparison with clinical and laboratory data.

Materials and methods. 63 patients with histologically confirmed PC and 52 patients with histologically confirmed BPH were under observation. The control group consisted of 30 practically healthy persons comparable in age. Determination of the relative level of IL-10 mRNA in PB samples was performed by real-time reverse transcription polymerase chain reaction.

Results. Both in patients with PC and in patients with BPH, a statistically significant decrease in the level of IL-10 mRNA in the PB of patients was observed in comparison with the control. In PC, the lowest levels were found in patients with a prostate-specific antigen (PSA) concentration above 10 ng / l and with a prostate volume of more than 50 cm3. Differences in the level of IL-10 mRNA at T2 and T3 stages and at different testosterone concentrations were not statistically significant, although there was a pronounced downward trend in prognostically unfavorable cases. Patients with BPH had a relative level of IL-10 mRNA, which was statistically significantly higher than in patients with PC. At PSA concentrations above 10 ng / mL, the level of IL-10 mRNA was also lower than at its lower concentrations.

Conclusion. In patients with cancer and BPH, a reduced level of IL-10 mRNA was found in the PB. The decrease is more pronounced in the unfavorable course of diseases and, apparently, is a consequence of the instability of IL-10 mRNA at the post-transcriptional level.

Background. Experimental animal models of psoriasis helped to clarify the functions of inflammatory mediators, to reveal the contribution of innate or adaptive immune mechanisms, keratinocytes to the development and maintenance of inflammation in psoriasis.

Aim. To study the subpopulation composition of immune cells of blood, skin, lymphoid organs and compare two methods of isolation of cells from the skin.

Materials and methods. The study included 46 mice of the C57BL / 6 line, which were divided into 2 groups: experimental (n = 24) to reproduce a model of acute psoriasis-like dermatitis using imiquimod cream 5 % (62.5 mg / cm2 / day / mouse, 7 days) and control (n = 22). The severity of skin inflammation was assessed on a point scale. On the 7th day, the skin, spleen, lymph nodes, and thymus were examined. To isolate cells from the skin, the method of spontaneous migration and enzymatic dissociation using collagenase was used. The assessment of the subpopulation structure of mononuclear cells (MNCs) was carried out by flow cytometry using monoclonal antibodies against the corresponding antigens (CD3, CD4, CD5, CD8, MHC class II, TCRyδ, CD38, CD80, CD83, CD86, TLR2). Statistical processing was carried out using the winMDI 2.8 software package.

Results. It has been shown that both methods of isolation of skin cells are applicable for immunophenotyping of γδ T-lymphocytes, CD86⁺, CD83⁺, CD83⁺CD86⁺ dendritic cells. A decrease in TLR2 expression on blood cells and an increase in lymph node and skin cells were revealed. There was a marked increase in the number of CD38⁺ in the lymph nodes, thymus, and an increase in γδ T-lymphocytes in the lymph nodes and blood. The infiltration of γδ T-lymphocytes, CD8⁺ is shown in the skin and CD38⁺ cells.

Conclusion. Acute psoriasis-like inflammation of mice was accompanied by an increase in the number of γδ T cells in the blood, lymph nodes and skin. Infiltration of the skin by CD8⁺ and CD38⁺ cells was observed. Both methods of cell isolation – the method of spontaneous migration and the method of enzymatic dissociation proved to be applicable for further immunophenotyping.

Backgraund. Pneumolysin (Ply) is a hemolytic toxin of Streptococcus pneumoniae (S. pneumoniae) expressed by all strains of pneumococci. The use of sandwich enzyme-linked immunosorbent assay (ELISA) can be a simple, fast and effective way of its qualitative and quantitative determination in biological fluids.

Aim. To develop and evaluate the specificity of sandwich ELISA test system for qualitative and quantitative determination of recombinant Ply (rPly) of S. pneumoniae.

Materials and methods. Immobilized on the solid phase rabbit’s polyclonal antibodies (pAbs) to rPly were used as recognition antibodies in sandwich ELISA. The studied antigens were added to the pAbs (rPly). The reaction was manifested by using detecting mouse monoclonal IgG1 (rPly) – antibodies conjugated with horseradish root peroxidase. The specificity of the test system was evaluated when using recombinant α-hemolysin (rα-Hly) and water-soluble S. aureus antigens as reference preparations.

Results. Using sandwich ELISA, rPly was detected at a concentration of 0.15 µ / ml. The test system was characterized by specificity, which was confirmed by the absence of reaction with recombinant rα-Hly of Staphylococcus aureus (S. aureus). Reference preparations of water-soluble surface antigens of S. aureus strains No 209, 1986,1991 and Cowan I gave a false positive reaction due to the presence of protein A (SpA) in their composition, a thermostable surface protein expressed by many strains of staphylococci capable of binding immunoglobulins via Fc-fragment or Fab fragments of the V3H domain of the B cells receptor. A negative reaction was obtained with antigens from the S. aureus wood 46 strain, which does not have the spa gene encoding SpA expression. The presence of protein A in preparations of water-soluble S. aureus antigens was confirmed in the ELISA inhibition assay.

Conclusion. Sandwich ELISA has been developed for qualitative and quantitative determination of S. pneumoniae Ply. The conducted studies have confirmed the specificity of the test system.

Background. Increasing the solubility of new pharmacologically active substances is one of the main tasks of pharmacy. It becomes even more relevant in the field of creating dosage forms for difficult-to-solubilise substances with a new mechanism of action, as in the case of 3-hydroxyquinazoline derivatives, which have shown in vivo experiments the ability to activate tumour cell death by ferroptosis. This includes OVF-009 – an original domestic substance. Due to the solubility of this compound in oils, the technology of its solubilisation using modified castor oil (Kolliphor® ELP, BASF, Germany), approved for parenteral use, was proposed.
Aim. To create a model dosage form for a new hydrophobic quinazoline derivative in order to further evaluate the spectrum of its antitumour activity in in vivo experiments.
Materials and methods. OVF-009, Kolliphor® ELP, 95 % ethanol, Kollidon 17 PF (BASF, Germany), sodium hydroxide, phosphate buffer, water for injection; spontaneous micelle formation, ultrasound, potentiometry, dynamic light scattering, electrophoretic method, viscometry method, etc.
Results. The main properties of micelle-forming solubiliser Kolliphor® ELP and its aqueous solutions were considered; 10 model solutions OVF-009 based only on Kolliphor® ELP and with addition of ethanol and Kollidon 17PF in different concentrations were obtained; the quality of the obtained compositions was evaluated by parameters – appearance, solution transparency, pH, stability over time and tolerability by laboratory animals. As a result, two formulations prepared on phosphate buffer were chosen: formulation No 5, which contains 10 % Kolliphor® ELP and additionally 10 % polyvinylpyrrolidone, and formulation No 8, in which in addition to the main solubiliser 10 % ethanol and 16 % polyvinylpyrrolidone were added. Both formulations have neutral pH, can be stored for 24 h and are tolerated by laboratory animals.
Conclusion. The selected model compositions for solubilisation of the substance OVF-009 on the basis of Kolliphor® ELP allowed to provide the concentration of the active substance in the solution suitable for further biological experiments on animals.

Introduction. when developing the composition of drugs, an actual direction is the use of computer modeling methods, including the methods of molecular dynamics (MD), which significantly expanded the possibilities of chemistry, providing spatial and temporal resolution that is inaccessible in experiments.
Aim. To simulation of the release of vinpocetine from sodium alginate with a shell of chitosan into solvent media by the method of MD to determine the characteristics of computer simulation, which makes it possible to obtain microcapsules with desired biopharmaceutical properties. 
Materials and methods. To simulate the release of vinpocetine from sodium alginate with a shell of chitosan, the MD method in the GROMOS 54a7 force field was used using the Gromacs 2019 program. Using the HyperChem 8.0.1 program, the molecules of the components of the simulated systems were constructed. The models were parameterized using the Internet service Automated Topology Builder (http://atb.uq.edu.au/).
Results. Based on the results of MD modeling, the van der waals interaction energies of vinpocetine with sodium alginate (alginic acid), with chitosan (chitosan-cation) and with a solvent in terms of 1 molecule of vinpocetine were calculated. The fractions of vinpocetine molecules not bound to the polymer were also calculated. It has been established that the average values of the energy of the van der waals interaction between vinpocetine and the solvent in an acidic medium are lower than in a neutral medium. Also, in an acidic environment, in contrast to a neutral environment, a slight release of vinpocetine is observed.
Conclusion. In the course of the experiment, it was found that at pH 2.0, chitosan dissolves in an aqueous medium and a slight release of vinpocetine from alginic acid into an aqueous solution of chitosan is observed (the average proportion of vinpocetine molecules not associated with sodium alginate (alginic acid) and chitosan is 2.16 ± 2.33 %), the release of vinpocetine into water at pH 6.8 is not observed.