Malignant tumors are known to have complex mutational profiles and harbor concurrent alterations in many somatic genes. The inher ent genetic heterogeneity is a critical determinant of cancer cells. The term of neoantigens was introduce to emphasize that these antigens are specific for cancer cells and are absent in normal tissue. Neoantigenes are highly immunogenic and at present are considered to be the target molecules in cancer therapy. Although the neoantigens have been known for a long time, their study and use became possible with the increase in the availability of mass cluster sequencing to detect all mutations in tumors and bioinformatic algorithms predicting what mutated peptides will be highly affine to human leukocyte antigen autologous molecules with subsequent activation of the immune response.

The purpose of this work is to demonstrate the existing possibilities and methodology of mathematical modeling of the freezing phase in the technology of lyophilization of vials filled with a solution containing a pharmaceutical substance and auxiliary substances. The freezing process is crucial for the subsequent stages of primary sublimation and the drying stage. So it is at the stage of freezing that ice crystals are formed and a certain microstructure forming the pores for subsequent stages and influencing the speed of all stages in the future. The developed methods allow estimating the kinetics of cooling, freezing, calculating the sizes of ice crystals and determining the permeability of the sublimation layer. Of course, for these calculations it is necessary to know the conditions and values of the variables, calculated and measured during the pilot freezing. However, modeling allows to reduce the time of development and optimization of the process, increases the speed of transfer of the technological process to other equipment. In this review, formulas are analyzed, the thermal conductivity equations for each cooling zone, equations for estimating the crystal size and pore size in the dried layer, and calculating the permeability of the sublimated layer. The article draws conclusions about the perspective of modeling methods for the freezing phase, examines the equations used in modeling and demonstrates the model of supercooling, crystal formation and other mass and heat exchange processes during the cooling and freezing stage. The calculations presented in this paper are confirmed by references to experimental data and have great practical value.

Brain metastases are very common in non-small cell lung cancer. The condition of these patients is complicated and difficult to treat, and adverse reactions following treatment can affect the nervous system, which severely reduces quality of life. In this review we discuss the methods in use from local therapies such as whole brain irradiation, stereotactic radiosurgery and central nervous system surgery to systemic therapy. We also highlight drugs for targeted therapy of non-small lung cancer cells metastasis in the brain that have been effective in the treatment of metastases in central nervous system.

Introduction. Approaches based on the principles of a targeted therapy are considered a promising strategy that is capable to improve the effectiveness of treatment for bladder cancer (BC) patients.

The purpose of the study was to establish an orthotopic xenograft model of human BC in mice and to prove its suitability for experimental examination of drugs targeting the epidermal growth factor receptor (EGFR).

Materials and methods. The objects of the study were ectopic subcutaneous and orthotopic human BC xenografts established using EJ and 5637 human BC cell lines. The growth of orthotopic xenografts in vivo was assessed by magnetic resonance imaging. Tumor tissues were investigated using histological and immunohistochemical techniques.

Results. It was shown that EJ and 5637 xenografts exhibit a good reproducibility, a sufficient blood supply of the tumor tissues, a high level of EGFR expression, and different pattern of a subcellular receptor localization. Implantation and subsequent proliferation of human EJ or 5637 cells in the murine bladder mucosa presumably results in muscle-non-invasive tumor formation.

Conclusions. The EJ and 5637 xenograft models can be useful for investigation of the efficacy of EGFR-targeted biotherapeutic treatments.

Introduction. The given data of fundamental studies of apoptosis processes in B-cell lymphocytic leukemia (B-CLL) testifies about the complexity and variety of mechanisms affecting the kinetics of normal cells and tumor lymphocytes in this disease. It is important to study the severity of clinical manifestations of the disease depending on the expression of the genes that modulate apoptosis.

The purpose of the study is to compare the activity of genes encoding apoptosis modulators, the cell cycle and cancer-testicular PRAME protein with clinical manifestations of the disease in primary patients with B-CLL.

Materials and methods. The level of expression of the proapoptotic genes FAS, TRAIL, TNFR2, DR4/5 and DR3, as well as the HSP27, XIAP genes, blocking apoptosis was determined in 23 patients with newly diagnosed chronic B-CLL. In addition, expression of genes TP53 and P21 and cancer-testis gene PRAME are tested.

Results. According to the multivariate regression analysis, the FAS gene expression in the onset of the disease had the greatest impact on the clinical characteristics of the disease. In this connection, the patients were divided into groups with normal (group) and low gene level (group II). A low level of FAS expression (Me 387 %) was associated with stage II disease (p = 0.03), a large number of lympho cytes (p = 0.001), fewer erythrocytes (p = 0.08), and a lower level of TNFR2 gene expression (p = 0.08), high level of expression of XIAP, HSP27, P21. Overall, the anti-apoptotic potential in Group II patients was higher, which was accompanied by more pronounced clinical manifestations of the disease.

Conclusions. The increased anti-apoptotic potential of tumor lymphocytes in newly diagnosed B-CLL is accompanied by a larger tumor mass and greater clinical and hematological manifestation of the disease.

Background. Clinical studies have shown that the most oncological patients have impaired immunological reactivity. Monitoring of the immune system status of patients with solid tumors includes a subpopulation analysis of immunocompetent cells. It is based on the study of expression of surface antigens and intracellular structures, and an analysis of the functional activity of immunocompetent cells. Today two, three, and even four-color reagents are used for subpopulation analysis of lymphocytes. Fluorescent conjugate with monoclonal antibodies (MAB) of the ICO series and fluorescent dyes Imd-306, Imd-506 (cyanine) and phycoerythrin (a group of phycobiliproteins) was received and characterized in N.N. Blokhin National Medical Research Center of Oncology. These conjugates have proved themselves in one and two-color analysis of subpopulations of peripheral blood lymphocytes by flow cytometry.

The purpose of this study is to assess the possibility of sharing several immunofluorescent probes (IFP) on the basis of different Mab series of ICO and dyes FITC, Imd306, Imd506 and PE for three-color analysis of lymphocyte subpopulations of donors and patients with oncological diseases by flow cytometry method.

Materials and methods. A panel of immunofluorescent probes (IFP) CD3FITC, CD4PE, CD8-Imd506, CD4-Imd306 was used in the work. Probes are obtained with MAB (ICO series) and fluorophores FITC, PE, Imd-306, Imd-506. The specific activity of the probes was evaluated in the immunofluorescence reaction by flow cytometry. Donor’s peripheral blood was used to optimize the conditions of the immunofluorescence reaction. Clinical approbation of the obtained sets was carried out on 2 groups of patients with oncological diseases before and after surgical treatment. The first group included 64 patients with cancer of the oral mucosa. The second group consists of 35 patients with ovarian cancer.

Results. We have shown that for the three-color analysis of human lymphocytes by flow cytometry, it is recommended to use sets of labeled MAB: CD3-FITC (10 μg/ml) + CD4-Imd306 at a concentration of 5–10 μg/ml; CD3-FITC + CD4-PE at a concentration of 20 μg/ml and CD3-FITC + CD8 Imd506 at a concentration of 10 μg/ml. The final concentration of immunoglobulin in the solution of IPS is indicated. It was shown that even before the beginning of treatment in patients with cancer of the oral mucosa the subpopulation CD3+ CD4+ content was significantly lowered in the structure of T cells, and the total level of CD8+ lymphocytes significantly exceeds the parameters of the donor group. Analysis of linear CD markers of lymphoid cells in patients with ovarian cancer revealed a significant decrease in the total number of CD3+ T-lymphocytes in comparison with donors. There was no evidence of the effect of surgical treatment on the subpopulation structure of lymphoid cells.

Conclusion. Combinations of fluorescent probes CD3-FITC/CD4-Imd306/CD8-Imd506 and CD3-FITC/CD4-PE/CD8-Imd506 can be used for trichromatic analysis by flow cytometry of the lymphocyte subpopulations of both healthy donors and cancer patients. Comparative studies of the specific activity of combinations of IFP based on Mab ICO series and reference commercial test systems (BD Biosciences) on lymphocytes of healthy donors and oncological patients (ovarian cancer, oral mucosa cancer) showed comparable results. The application of the obtained probes made it possible to reveal violations of the subpopulation structure of peripheral blood lymphocytes in patients with ovarian cancer and cancer of the oral mucosa, in particular, CD3 – /CD8+, CD3+/CD4+, CD3+.

Background. Aranosa, a drug of nitrosoalkylureas class, was developed and studied at N.N. Blokhin NMRCO, Russia, and at present it is produced by N.N. Blokhin NMRСO branch “Naukoprofi”. One of the stages of the drug standardization is the development of an assay technique for quantitative determination of active substance in the final drug dosage form.

Objective. Development and validation of an assay for quantitative determination of Aranosa in the dosage form.

Materials and methods. The study used “Aranosa, lyophilisate for solution for injection 0.5 g”; Аranosa, substance-powder (N.N. Blokhin NMRCO branch “Naukoprofi”); polyvinylpyrrolidone low-molecular medical; sorbic acid. Method: spectrophotometry.

 Results. An assay was developed for quantitative spectrophotometric determination of Aranosa in the dosage form “Aranosa, lyophilisate for solution for injection 0.5 g”. Validation was performed to prove reliability and accuracy of the obtained results. The assay was evaluated by validation characteristics, such as specificity, linearity, trueness, precision.

Conclusions. The developed assay is provided with trueness, repeatability, precision, and linearity and can be used in the range of 80– 120 % of nominal Aranosa content in the dosage form.

Introduction. The work is devoted to preclinical toxicological research on laboratory animals tsifetrilin – a drug, an analogue of the hypothalamic hormone somatostatin, selected according to the results of studying hormonal, cytotoxic and antitumor activity.

The purpose was preclinical toxicological studies of the drug’s tsifetrilin – an analogue of the hypothalamic hormone for the treatment of malignant hormone – dependent diseases.

Materials and methods. Studies were carried out on 80 healthy mice hybrids (CBA × C57Bl/6J) F1 males and females, 100 non-inbred healthy mongrel male and female rats and 5 breed dogs English Beagle males and females. Preclinical toxicological studies were carried out using tsifetrilin granulate, which contained 57.3 mg of Citrate in 1 g of granulate. As a solvent, 1 % starch paste was used. In the experiments to study acute toxicity were administered orally once tsifetrilin: mice at doses of 100, 200, 300 and 600 mg/kg, to rats in doses of 100, 200, 400 and 500 mg/kg. The period of observation of the animals was 30 days. In experiments on chronic toxicity, the drug was administered orally 15 times a day: rats – total doses of 30 and 300 mg/kg; dogs – total doses of 124.5 and 622.5 mg/kg. The observation period in studies in rats was 45 days, in studies on dogs – 60 days.

Results. As a result of the study of acute toxicity in mice and rats in females and males when tsifetrilin was administered at the maximum possible concentration and the maximum possible volumes, the drug did not cause death of animals, did not affect the general condition of the animals, caused no external toxicity, animals. In the study of chronic toxicity of tsifetrilin in rats and dogs with daily oral administration for 15 days, no death was observed in all the doses studied, and the drug did not cause any external toxicity. The limiting type of toxicity is not established, since the drug causes minor morphofunctional changes of varying degrees of reversibility in virtually all organs and systems of the body of rats and dogs. Based on the experimental data on functional and morphological changes in organs and tissues, it was shown that females are more sensitive to the preparation than males.

Introduction. The existence of the active metabolite (amino acid) residue in the of an indolocarbazole molecule changes physical-chemical and pro-medicinal properties of aminoacid derivatives glycosides of indolocarbazole. The computer method has earlier foretold low probability of their cytotoxic activity in vitro that was confirmed in the MTT-test on 5 lines of tumor cells. The same computer method predicted significant probability of antineoplastic activity of aminoacid derivatives glycosides of indolocarbazole in vivo that demands the experimental check.

 Purpose of the study – assessment of aminoacid derivatives glycosides of indolocarbazole as potential antitumor medications.

Materials and methods. The research of antineoplastic activity of aminoacid derivatives glycosides of indolocarbazole was performed on mice tumoral models – cervical cancer СС5. Abdominal injections were made to CBA/Lac mice 5 times a day with 24 h interval. Observation of animals was continued till their death. The antineoplastic effect of medicines was estimated according to tumor growth inhibition, increase in life expectancy of experiental mice in comparison with control animals.

Results. The optimum dose for this number of compounds, equal to 100 mg/kg is titrated. The antineoplastic activity of aminoacid derivatives glycosides of indolocarbazole on the model СС5 is estimated.

Conclusions. On the basis of the obtained data the expanded research of antineoplastic properties of the selected 5 aminoacid derivatives glycosides of indolocarbazole is supposed to perform in vivo.

The purpose of this investigation was to evaluate the efficacy of correcting the LFA-1 and Mac-1 leukocyte integrins expression on peripheral blood cells as well as IL-6 and IL-10 serum levels using liquid form multiphytoadaptogene preventive application for hepatocarcinomas incidence suppression and life-span increase of CBA inbreed mice.

Materials and methods. The study was carried out on 439 males of inbred CBA mouse strain (subline CBA/LacY). The experimental mice received 10 % liquid form multiphytoadaptogene complex (MPAC) solution in drinking water during the first month of life including the final time period of liver differentiation (preventive administration). MPAC is the standardized herbal formula composed of 40 plant extracts components including adaptogenes Panax ginseng, Eleutherococcus senticosus, Rhodiola rosea, as well as compounds with phenolic structure (flavonoids, triterpene glycosides, etc). Anti-mutagenic, anti-oxidant, immunomodifying activities of MPAC were demonstrated. The CD11a and CD11b antigens expressions on peripheral blood cells were analyzed by indirect immunofluorescence reaction, serum cytokines IL-6 and IL-10 concentrations were determined by enzyme-linked immunosorbent assay as well as the liver tissue morphology were analyzed at the age of 4, 8, 22 months. Tumor incidence and volumes were evaluated at the age of 8 and 22 months. Motor activity and physical status (body weight, coat state) were estimated in the ontogenesis. The average life-span and survival median was analyzed by the Kaplan–Meier method. Statistical analysis was performed with the program STATISTICA 6.0.

Results. Inhibited expression of LFA-1 and Mac-1 leukocyte integrins on peripheral blood cells as well as elevated IL-6 and IL-10 serum levels, high incidence of liver tumors (100 % of mice-males), their high number and big sizes in the late period of postnatal ontogenesis in CBA high-cancer inbrede mice, also as life-span not reaching two years was demonstrated. At the same time plural tumor-infiltrating limphocytes were not found. Short-term MPAC preventive administration (during the first month of mouse life occupying the final differentiation period of liver tissue genetically predisposal to hepatoms incidence) revealed the long-term heterotypic adhesion molecules leukocyte integrins LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18) up-regulated expression, providing the contact interactions of immune effectors and cancer cells. The events shown is able to enhance the anti-tumor activity of immune reactions including spontaneous hepatocarcinomas lymphocytes infiltration and destruction of tumor tissue. As a result, the hereditary tumors incidence, their number and sizes were reduced as well as life-span and life quality of animals were improved.

Conclusion. Up-regulated leukocyte integrins expression on peripheral blood cells, reduced IL-6 and IL-10 serum levels accompanied by tumors lymphocyte infiltration and destruction using MPAC preventive administration is essential for tumor incidence down-regulation. The herbal formula with multiple components is capable to controle anti-tumor immune reactions, as well as the life-span and life quality of high-cancer animals. Hence, the regulation of adhesion violations involved in tumor incidence can be cancer preventive.

Introduction. Lymphocytes blast-transformation reaction (RBLT) is necessary for patient’s inspection with immunologic infringements. It is applied in the different fields of medicine to identification of a sensitization to antigens (allergens).

Research objective – to modify and automate RBTL for application in routine laboratory practice.

Materials and methods. RBTL, antigens (allergens). Software: Windows 7; Intel Pentium G4500 CPU. Digital CMOS video camera, Mikmed-6 microscope.

Results. We analyzed difficulties when using RBTL in laboratory allergological practice. We created program files for automation of RBTL.

Conclusion. We adapted and automated RBTL for laboratory diagnostics using.