Influence of combined transplantation of multipotent mesenchymal stromal cells and stellate liver cells on its morphofunctional state after partial hepatectomy

The aim of the study was to study the changes in the morphofunctional state of the liver after the combined transplantation of multipotent mesenchymal stromal (MMSC) and stellate liver cells (ZCP) in animals with partial hepatectomy.Materials and methods. The MMSC culture was isolated from the placental chorion of 5 laboratory animals f female mice aged 3-4 months, weighing 22-23 g, gestation period of 18 days. The mononuclear fraction of the cells was obtained by sequential mechanical and enzymatic treatment of the placental tissue. The cultivation of MMSCs was carried out in a CO2 incubator at a temperature of 37 0C with a carbon dioxide content of 5% and a humidity of 90%. ZCP was isolated by collagenase-pronase perfusion of the liver, followed by cell separation in the histodense density gradient. MMSCs of the 3rd passage were introduced, and ZKP was introduced immediately after isolation. MMSCs at a dose of 4 million cells/kg and ZCP at a dose of 9 million cells/ kg were used for transplantation to labotory animals. The cells were injected 1 hour after partial hepatectomy. The biochemical parameters of peripheral blood and morphometric parameters of the liver were evaluated on the 3rd, 7th day after the administration of MMSC.Results. As a result of the study, it was found that the combined transplantation of MMSC and ZCP after partial hepatectomy leads to the restoration of the level of enzymes that characterize cytolysis and cholestasis, normalization of the protein-synthetic function of the liver, normalization morphometric parameters of the liver. A significant mechanism for restoring the morphofunctional state of the liver can be considered the influence of transplanted MMSCs and ZCP on the cell repair system, which leads to a decrease in the severity of programmed cell death of hepatocytes and the level of pathological mitoses.Discussion. Combined transplantation of MMSCs and HCP after partial hepatectomy leads to an increase in the level of hepatocyte growth factor (HGF), thus contributing to an se in the mitotic activity of hepatocytes and the restoration of liver mass. The transplanted cells also, by increasing the activity of DNA repair enzymes of the PARP family, lead to a decrease in the level of pathological mitoses, inhibition of their programmed cell death.Conclusions. The conducted studies demonstrate the ability of combined MMSC and PCR transplantation to restore the morphofunctional state of the liver after partial hepatectomy and provide the basis for conducting pilot clinical studies

Influence of Chronic Ionizing Radiation on the Variability of Mitotic Activity of Pinus sylvestris L. Tissues

The development of nuclear power engineering will increase in the future, due to the continued likelihood of radionuclides entering the environment. The relevance of studying the genetic material of conifers as a reliable bioindicator was revealed. It is known that under the influence of ionizing radiation there is an accumulation of genetic abnormalities in pine (Pinus sylvestris L.) progeny. The stages of gametogenesis, zygotes and young seedlings of plant organisms are most sensitive to ionizing radiation. It is important to determine the variability of the mitotic index (MI) of tissues of Scots pine (Pinus sylvestris L.) progeny in areas with different levels of ionizing radiation, including in the exclusion zone. The exposure dose rate was measured at the soil level and 1 m from the ground surface. Germination of seeds was carried out in a thermostat on wet filter paper. Roots of seedlings 0.5–1 cm long were fixed in a mixture of 96 % ethanol and glacial acetic acid. Root staining was carried out in a solution of acetocarmine. Tissue softening was carried out with a strong chloral hydrate solution. The total number of dividing cells, as well as pathological mitoses (PM) were counted on squash preparations using a microscope. The mitotic index and the duration of the phases of mitosis were determined. An increase in the level of radioactive contamination increases the cell division rate in prophase, anaphase, and telophase, but decreases the duration of metaphase, and also changes the relative duration of mitosis phases. With an increase in the exposure dose rate, the number of pathological mitoses naturally increases. The spectrum of mitosis disorders is represented by various abnormalities of the chromosome apparatus in anaphase: exit and lagging of chromosomes, bridges. Herewith, the number of anaphases with bridges increases significantly with simultaneous exit and lagging of chromosomes.

Cytogenetic polymorphism of seed progeny of walnut (Juglans regia L.) during introduction in the Central Chernozem Region

We studied the cytogenetic characteristics of the seed progeny of the walnut (Juglans regia L.) trees introduced to and grown within the territory of the Central Russian Upland. Three seedling groups with polymorphic cytogenetic characteristics were revealed: mutable (with a high level of pathological mitoses), low mutable (with a low level of cytogenetic disturbances), and intermediate groups. Cytogenetic characteristics (mitotic activity, parts of cells at various stages of mitosis, the level and spectrum of pathological mitoses, sizes of nucleoli and the spectrum of their types, the occurrence of cells with a persistent nucleolus in the stages of meta-, ana-, and telophase) in each of the selected groups were described; homeostatic mechanisms at the cellular level were discussed. The sizes of polymorphic groups were established. The small number of seedlings with a high level of cytogenetic disturbances (7.5%) and the predominance of seedlings with medium (70%) and low (22.5%) values of pathological mitoses indicated a high degree of adaptation of the introduced walnut mother trees to the environmental conditions of the Central Russian Upland. Predictors for assigning any seedling to one of the selected model groups (mutable or low mutable) were established using ROC analysis methods. The obtained data on the qualitative and quantitative polymorphism of cytogenetic characteristics can be used for the development of recommendations for improving the system of seed production and the selection of new forms of walnut in the Central Chernozem Region.

Effect of Beta-lactam Antibiotics on Microscopic Parameters in the Allium-test

The work examines the effect of beta-lactam antibiotics (cefotaxime, ampicillin, augmentin) on the pathology of mitosis in the Allium–test. Research methods: Allium–test, cytogenetic analysis, statistical analysis. It was established that the use of individual tested beta-lactam antibiotics increases the percentage of pathological mitoses in the cell by 1.8–3.3 times compared with the value in the control. With the combined use of cefotaxime and Augmentin, synergism appeared, as a result, the value of mitosis pathology turned out to be at the level of the number in the control; minimally represented pathologies indicating damage to the mitotic apparatus. It was revealed that all three beta-lactam antibiotics tested had a pronounced statmokinetic effect. At the same time, with the joint use of cefotaxime and Augmentin, k-mitosis was not registered in dividing cells. Comparison of the spectrum of pathological mitoses in the variants of the experiment showed that the pathology ‘chromosome runaway/backlog’ in anaphase of mitosis dominates in all variants. An increase in the concentration of Augmentin and ampicillin caused the suppression of pathological processes in onion meristematic cells, a decrease in PM values was observed. An increase in Augmentin concentration does not affect the composition and spectrum of pathological mitoses; ampicillin has a decrease in the level of most of the recorded pathologies of mitosis.