—A review of the information published in the scientific literature and relating to estimation of spontaneous levels of translocations detected by various methods of FISH-staining of chromosomes in the human peripheral blood lymphocyte cultures is presented. Various authors using single, two, and three-color variants are generally indicated by a significant increase in the frequencies of these chromosome aberrations with an increase in the donor age and the absence of the influence of gender. An increase in the translocation yield was recorded for persistent smokers and, apparently, alcoholics. It was recognized as impossible to divide the effect of the race and the territorial position of research laboratories. At the same time, countries can differ significantly in terms of the lifestyle of the population, the background impact of ionizing radiation, and the medical services used, in particular, in terms of the volume of implemented radiation diagnostic research. It limits the use of spontaneous damage levels of chromosomes determined by international studies and indicates the desirability to have relevant data for individual populations. At the same time, it is necessary to state the limitation of similar information about the frequencies of translocations detected with multicolored FISH-staining in the control groups of people.

Objective: to compare the frequencies of translocations detected by one-color and three-color FISH painting in the long term (24 years) after local over-irradiation (more than 100 Gy) of a patient during radiation therapy for breast cancer. Material and methods: The material for the cytogenetical study were the cultures of venous blood lymphocytes of this patient. Cultivation of lymphocytes and preparation of chromosome preparations were carried out using variants of standard methods. Chromosomes were stained separately with one-color or three-color DNA probes for 1, 4 and 12 pairs. In both cases, 1000 metaphases were analyzed. Results: In the present study, when using the one-color FISH-method, the frequency of detected reciprocal translocations was 0.5 per 100 analyzed cells, while with tricolor painting they were 2.2 times higher (1.0 per 100 metaphases). When using the available linear-quadratic dose dependence, the average dose to the whole body was estimated at 0.28 Gy, which is a consequence of the local nature of radiation damage, the death of lymphocytes in the zone of high-dose radiation damage and the obvious impossibility of their division. Conclusion: The study of the frequency of chromosomal translocations in the long term after local overexposure during therapy in a patient for breast cancer using a three-color FISH-analysis is a more sensitive method in comparison with a one-color variant of FISH-painting.

This article describes the process of developing a tissue-engineered structure that meets the biocompatibility and biodegradation parameters necessary for replacing cartilage tissue defects. The study was carried out using 3D bioprinting technology, which represents a promising research direction in the biomedical field. It is known that, due to the specifics of its structure, cartilage tissue is not capable of complete regeneration of damage. The methods currently used for treating arthrosis are associated with a number of limitations and disadvantages, which makes research aimed at developing alternative methods for arthrosis treatment particularly relevant. The development of tissue-engineered structures by 3D bioprinting requires the materials not only certified for medical use but also exhibiting biocompatibility and biodegradation properties. Polylactide (PLA) and sodium alginate satisfy the above requirements; moreover, their availability and economic affordability make them one of the most popular materials for 3D bioprinting.

— The results of cytogenetic analysis using mainly three-color FISH staining of peripheral blood lymphocyte chromosomes in a single patient exposed to irradiation as a result of the accident at the Chernobyl nuclear power plant in a dose initially estimated from the frequency of dicentrics (3.2 Gy) are presented. The material was obtained in a total time interval of 9053–12 554 days (24.8–34.4 years) after the radiation effect. Not always together, the sets of DNA probes to {1, 4, 12} and {2, 3, 8} pairs of chromosomes were used at all times. For the entire period of observation, the genomic frequency of translocations (a total of complete and incomplete) did not change statistically significantly. The retrospective dose estimations were made using previously obtained in vitro dose–effect dependences and ranged from 1.98 to 2.44 Gy and from 2.14 to 2.67 Gy when analyzing the metaphases of all cells and only stable cells, respectively. The problem of accounting for clonal cells with chromosome translocations is discussed.

Purpose: To evaluate the frequency and spectrum of chromosome aberrations under X-Ray exposure at doses of 80, 250, and 1000 mGy in a human multipotent mesenchymal stromal cell (MMSC) cell line during long-term cultivation. Material and methods: MMSCs were isolated from human gingival mucosa by an enzymatic method and cultured in a serum-free medium. The presence of surface antigens was determined using the method of flow cytometry. The ability of the cell line to differentiate in the osteogenic, adipogenic, and chondrogenic directions was studied using induction media. Authentication was performed by genotyping of polymorphic STR loci, cytogenetic analysis was performed by multicolor fluorescent in situ hybridization (mFISH). Irradiation was carried out on an X-ray biological unit RUB RUST-M1 (Russia) at a dose rate of 40 mGy/min, a voltage of 100 kV, and a current of 0.8 mA. Results: At the first passage after irradiation, a statistically significant increase in the frequency of non-clonal CA compared with the control was recorded at a dose of 80, but not 250 and 1000 mGy. At the late stages of cultivation, the average frequency of breaks per chromosome in the group of non-irradiated cells did not differ from the values obtained after irradiation at doses of 80, 250, and 1000 mGy (p > 0.05). However, in MMSCs irradiated at a dose of 80 mGy, damage occurred more often in pairs of chromosomes 6 and 10, and at a dose of 1000 mGy, in a pair of chromosomes 9. A single irradiation of MMSCs in vitro did not affect the growth and progression of MMSCs characteristic of the studied primary cell line, of clonal cells with chromosome translocations and monosomy X, but led to an increase in the representation of a clone with tetrasomy 8. The total number of random clones with chromosome translocations that arose de novo increased after irradiation at a dose of 1000 mGy. Conclusion: Minor fluctuations in the proportion of cells with non-clonal CA, depending on the dose received in the early stages after irradiation (passage 1–4), disappeared at the later stages of cultivation (passage 8–14). There were no differences in mean frequencies between irradiated and non-irradiated MMSCs, but after irradiation, damage to some chromosomes could occur more frequently than others. A single X-ray irradiation of MMSCs can promote the growth and progression of primary pathological cytogenetic clones, regardless of the dose received, as well as an increase in the total number of de novo cell clones with chromosomal translocations that have arisen. A single X-ray irradiation of MMSCs can promote the growth and progression of primary pathological cytogenetic clones, regardless of the dose received, as well as an increase in the total number of de novo cell clones with chromosomal translocations that have arisen.

This work presents the results of the use of three-color FISH-staining of chromosomes (DNA probes to 1, 4 and 12 pairs of chromosomes) for the cytogenetic study of peripheral blood lymphocyte cultures of four patients at different times after irradiation in various emergency situations. All victims had initial dose estimations: one by EPR signal in the tooth enamel (1.12 Gy) and three by the dicentric frequencies (1.9; 3.2 and 4.3 Gy). There was a strong significant linear correlation between the observed translocation frequencies and the initial dose estimations. Retrospective indication of the dose by translocation frequencies was carried out with the help of own dose-effect curve, the obtained on the basis of experiments with the irradiation of blood of healthy donors in vitro. At the same time, dose estimates in the long terms after irradiation in three of these patients with the greatest lesion were significantly lower than they were established earlier (1.26; 2.06 and 2.47 Gy). The coincidence of the initial and repeated results was observed only in one remaining patient.

This work presents the results of the use of three-color FISH-staining of chromosomes (DNA probes to pairs of chromosomes 1, 4, and 12) for the cytogenetic study of peripheral blood lymphocyte cultures of four patients at different times after irradiation in various emergency situations. All patients had initial dose estimations: one by EPR signal in the tooth enamel (1.12 Gy) and three by dicentric frequencies (1.9, 3.2, and 4.3 Gy). There was a strong significant linear correlation between the observed translocation frequencies and the initial dose estimations. Retrospective indication of the dose by translocation frequencies was carried out with the help of the dose–effect curve, obtained on the basis of experiments with the irradiation of blood of healthy donors in vitro. At the same time, the dose estimates in the long term after irradiation in three of these patients with the greatest lesions were significantly lower than they had been established earlier (1.26, 2.06, and 2.47 Gy). Coincidence of the initial and repeated results was observed only in one remaining patient.

Tumor necrosis factor (TNF) plays an important role in the pathogenesis of breast cancer (BC). The -308(G/A)TNF polymorphism allele -308A is part of the autoimmune ancestral haplotype AH8.1 associated with elevated serum sTNF levels. Previously, we showed a decrease in overall survival for carriers -308A/AH8.1-. The aim of the study was to assess the effect of haplotypes AH8.1 on the level of sTNF and clinical and morphological characteristics of BC patients. The study included 100 people with adjuvant radiotherapy treatment. The concentration of sTNF was determined by the ELISA. Additionally, 30 archival DNA samples of primary BC patients carrying the allele -308A with a known sTNF level were genotyped. Alleles -308(G/A)TNF and marker alleles of haplotype AH8.1 (HLA-A*01, HLA-B*08 and HLA-DRB1*03) were determined by allele-specific PCR. Based on the polymorphism of -308(G/A)TNF and marker alleles of haplotype AH8.1, three comparison groups were identified: -308GG carriers of the TNF gene independently of haplotype AH8.1 (76

Purpose: Study of chromosomal aberration yields with a three-color FISH method in the peripheral blood lymphocyte cultures of patients with breast cancer before and after local irradiation. Material and methods: Venous blood was obtained from 24 patients aged from 33 to 79 years which suffering from cancer of the left or right breast after the implementation of a sectoral or radical resection of an affected organ with subsequent local adjuvant distant radiation therapy (DRT) to the scar area. Also it could be preceded by adjuvant polychimotherapy (PCT). A linear accelerator of the Trilogy system based on the Clinac iX platform was used for local irradiation. SOD was installed at 50 Gy per 25 fractions (daily 2 Gy). venous blood sampling produced to PCT and/or DRT and after 1 and/or 3 months after irradiation. A set of DNA probes to 1, 4 and 12 pairs of chromosomes was used for three-color FISH staining. Results: Levels of translocations, dicentrics and acentrics (per 100 cells) in peripheral blood lymphocyte cultures of patients up to DRT did not differ significantly with and without PCT and were amounted to average 0.63 ± 0.10; 0.10 ± 0.04 and 0.17 ± 0.06, respectively. The frequency of these chromosome rearrangements significantly increased after radiation therapy by an average of up to 4.62 ± 0.46; 1.48 ± 0.19 and 0.95 ± 0.14. The differences between 1 and 3 months of taking material after irradiation were absent. One of the patients, unlike the rest, was cytogenetically examined 11 years after a similar local irradiation, and the frequencies of translocations and dicentrics in her lymphocyte culture were 4.56 and 0.19 (per 100 cells), respectively, while they were varied from 1.63 to 10.00 for translocations and from 0.27 (0 in one patient) to 4.06 for dicentrics for other patients. Conclusion: When using a three-color FISH method there was a significant increase in the frequencies of translocations, dicentrics and acentrics in peripheral blood lymphocyte cultures of patients after DRT of breast cancer. At the same time there were more translocations than dicentrics and, apparently, they remain at the initial level for a long time. The frequency of translocations per 1 cell with translocations in the peripheral blood lymphocyte cultures in most cases exceeds 1 after local DRT of the breast cancer. The correlation between average physical doses to the whole body and cytogenetical estimates in individual patients was statistically insignificant.

A cytogenetic study of cell lines of mesenchymal multipotent stromal cells (MMSCs) isolated from human bone marrow, adipose tissue, placenta, and gingival mucosa was carried out. Karyological analysis using two chromosome-staining methods (GTG and mFISH) showed a normal diploid karyotype of most MMSC lines up to the fifth passage of cultivation. Clonal chromosomal translocations were detected in 3 of the 22 investigated MMSCs, which is 14%. Analysis of 22 MMSC lines revealed a rather low frequency of cells with chromosomal aberrations (CAs) (within 5%). Among cells with chromosomal damage (3–5%) identified by mFISH analysis of 1440 metaphase plates, unstable chromosomal aberrations (fragments, dicentric chromosomes) account for about 65% of all damage, and stable (translocations) account for 35%. These results confirm the importance of cytogenetic studies of MMSC cell lines intended for medical purposes. Gathering data on chromosomal and karyotypic variability at different periods of cultivation will make it possible to determine the acceptable limits for the selection of genetically stable diploid MMSC cell lines in order to prevent undesirable consequences of their use for therapeutic purposes.

Purpose: to create a protocol for complex cryoconservation of blood vessels in polydimethylsiloxane to create tissue-specific matrices. Material and Methods. The material ofthe study was the iliac arteries from postmortem donors, seized in the multi-organ donation, rejected for unclaimed and disposed of later. Polydimethylsiloxane was used as a coolant (PDMS) with a viscosity of 5-25 centistokes (Spectroplast, Russia). Bench testing of cooling and heating ofthe blood vessel in PDMS, testing of heating and cooling of PDMS up to operating temperatures (-80°C) in different ways and variants of its thermal insulation, assessment of different cooling rates ofthe vessel during cryoconservation, modeling of 3 types of working chambers for cryoconservation, sterilization and defrosting of up to 10 vessels, modeling of cryoconservation and thawing of vessels in PDMS under different conditions, selection of substrates from silicone and plastic for uniform fixation of vessels which are of size or size in the working chambers or without them. Results. The developed protocol allows cryopreserved from 1 to 10 or more vessels of all basic sizes both immediately after removal and after transportation to the laboratory, to vary the cooling rate from 1.5 to 300°C/min and above, to be used with different levels of laboratory equipment, as with liquid nitrogen, dry ice, and only with a medical freezer. Conclusion. The developed protocol allows to apply it in cases of complex cryoconservation of blood vessels to create tissue-specific matrices and can be recommended for preclinical testing.