Cell and Tissue Biology

In order to improve the patient’s condition, the transfusion of blood products is used, however, there are problems with blood transfusion including infections, immune suppression, allergic reactions and anaphylactic shock-related to leukemia and plasma product. Cytokine can now be an appropriate choice to reduce the need for blood transfusion. Various types of cytokines have been studied in human clinical trials and some of them have a substantial function to decrease blood transfusion. In this study, we have investigated the important cytokines involved in reducing cells (Red blood cells and platelets) transfusion. First, we have investigated the cytokines involved in erythropoiesis and thrombopoiesis and covered some clinical studies about them. The second side effects of the administration of these cytokines have been addressed, and finally, we have suggested some recommendations about cytokine therapy.

The effect of X-ray radiation (0.1 Gy) on the expression of a number of genes and regulatory RNA (miRNA and long noncoding RNA) in human lymphocytes and T-lymphoblastic cells (Jurkat cells) was studied. One hour after cell irradiation with a low dose of radiation, lymphocytes displayed p53 expression and a decreased level of mature miR-27a and miR-181a having mRNA of gene p53 as target. These cells also had inhibited NFkB activity, which was found from reduced mRNA content of RhoAcdc42 and IL6 genes. By 4 h, their expression was normalized. Unlike in the case of normal cells, increased content of mRNA of NFkB gene (p65) and mRNA of its target IL6 gene was observed in Jurkat cells during this period. Repeated irradiation of cells with 5 Gy carried out after 4 h showed the radiation adaptive response (AO) according to the criterion of lymphocyte survival and its absence in Jurkat cells. The difference between groups of 5 Gy and 0.1 + 5 Gy in lymphocytes that survived after 20 h revealed common AO features (mRNA of p53 gene, NEAT1, miR-181a, miR-107). The results indicate the activation of various intracellular systems after the stress with low doses of radiation on lymphocytes and Jurkat cells. This approach can be used to optimize the efficacy of radiation therapy when preirradiation with a small dose of radiation increases the radioresistance of normal tissues surrounding a tumor.

The cerebellum is a part of the brain that is very sensitive to the toxic effects of general anesthetics. The aim of this work was to evaluate the morphological response of neurons and microgliocytes in all layers of the cerebellar cortex to prolonged (6 h) exposure of sevoflurane (general anesthetic). It was shown that, after prolonged exposure of Wistar male rats to sevoflurane, structural and functional rearrangement were observed in all layers of the cerebellar cortex. In the molecular and ganglion layers the total density of neurons decreased. The number of morphologically altered cells of the molecular layer and Purkinje cells increased to 250 and 300%, respectively, due to both direct toxic effects of the anesthetic and disruption of interneuron connections. In the granular layer, the total density of the neuron population did not change and the number of morphologically altered neurons did not increase significantly. The number of microgliocytes revealed immunohistochemically increased significantly, and activation in response to neuronal death was weakly present. The absence of excessive activation of microgliocytes after prolonged exposure to sevoflurane is a positive result, since neuroinflammatory mediators are produced to a lesser extent and neurons do not experience additional damaging effects from microglia.

The pharmacokinetics of two substances containing recombinant insulin-like growth factor 1 (IGF1) as an active substance was considered. The first substance (IGF₁) contains the actual recombinant IGF1, and the second (IGF₂) contains IGF1, which is translated from plasmid DNA encoding the IGF-1 gene for this protein. It was established that, with intramuscular administration of IGF₁, the delay time for its entry into the bloodstream is 1.5–2 h, while that with IGF₂ is 24–25 h. This indicates the presence of various mechanisms of accumulation of these substances in the systemic circulation. The maximum concentration of IGF₁ in the blood was determined 5 h after administration and that of IGF₂ was determined 125 h after administration. The maximum concentrations of these substances are comparable to each other. The concentration of IGF₁ in the blood decreases to its initial value 12 h after its administration, and the concentration of IGF₂ does so after 216 h. The clearance parameters (Cl) and elimination constants (Kel) of the considered substances also had significant differences, which confirms the presence of fast and slow dynamics of the decrease of their maximum concentrations after intramuscular injection. The different dynamics of the accumulation of substances in the blood and their elimination from the bloodstream after administration, as well as the different values ​​of the area parameters under the pharmacokinetic curve (AUC_t, AUC_∞) demonstrate that IGF₂ has been in the systemic circulation for a longer time than IGF₁. This is essential for the appearance and severity of pharmacodynamic effects.

Mechanically gated cation channels that are activated upon deformation of the membrane are key participants in the transmission of mechanical signals from the cell surface to cytoplasmic structures. It has remained unclear how mechanically dependent reactions involving ion channels in native cells are realized. In this study, we analyzed the development of the activity of single channels in K562 human myeloid leukemia cells in response to the delivery of a mechanical stimulus, this being stretching of the membrane area. The registration of ion currents using the classic versions of the patch–clamp method allowed revealing the functional clustering and interaction of various types of channels in the plasma membrane during mechanotransduction. In K562 cells, coupled activation of mechanosensitive calcium-conducting channels and calcium-activated potassium channels was detected. Real-time recordings of currents demonstrate that the calcium influx from the extracellular medium into the cytoplasm through mechanically gated channels activates potassium channels that are colocalized with them, which do not have their own mechanosensitivity. In experiments on K562 cells and transformed 3T3-SV40 fibroblasts, the preservation of the functional conjugation of the channels during their mechanically gated activation after the action of the microfilament destructor cytochalasin D was shown. The results suggest that clusters including potassium SK channels and Piezo1/2 proteins forming stretch-activated cation channels function in the plasma membrane of K562 and 3T3-SV40 cells.