S. aureus and E. coli change the force and work of adhesion between P- and E-selectins of endothelial cells and ligands of neutrophil granulocytes.

Thanks to the modification of the force spectroscopy method, when a neutrophil is fixed on the tip, and an endotheliocyte culture is grown on the substrate, the exact indicators of the adhesion force and adhesion work between cells have been investigated. The high variability of adhesion contacts in different donors associated with different expression profiles of neutrophils. It was found by flow cytometry that the EA.hy926 cell line actively expresses VCAM-1, as well as P- and E-selectin under the Staphylococcus aureus influence after 60 min of co-incubation. At the same time, the integral indicators of the adhesion force and adhesion work in the "neutrophil - endothelial cell" interaction were significantly inhibited by S. aureus in all studied donors. Since the VCAM-1 receptor is not involved in the adhesion bonds between neutrophils and endothelial cells, the suppression of the interaction is associated with the inhibition of P- and E-selectins, but direct receptors removal from the endothelial cells surface of the EA.hy926 cell line does not occur. Escherichia coli causes multidirectional effects in the system of interaction "neutrophil - endothelial cell", depending on the expression profile of the donor's neutrophils. However, the cumulative effect of interaction from all donors shows that in general, under the influence of E. coli, there is an increase in adhesion force and a suppression of adhesion work.

Serum Protein Corona Abolishes Changes in the Expression of Proinflammatory Genes Induced by Quantum Dots in Human Blood Mononuclear Cell.

We studied changes in the transcription of genes encoding cytokines (TNF, IL-6, IL-10, and IL-32), cell activation markers (ICAM1, CD38, Fas, and FCGRIII), ROS production catalyst (NOX2), autophagy (Beclin 1, LC3B, and p62) and apoptosis (BAX, BCL2) regulators in peripheral blood mononuclear cells upon contact with quantum dots CdSe/ZnS-MPA and CdSe/CdSeZnS/ZnS-PTVP. Up-regulation of TNF, ICAM1, Fas, p62 mRNA and down-regulation of the FCGRIII and NOX2 mRNA in response to the presence of quantum dots were revealed. The formation of serum protein corona on the surface of quantum dots abolished this effect.

Differences between Neutrophilic Granulocytes and Lymphocytes in Fixation of Quantum Dots of Different Composition.

Studies by flow cytometry, scanning electron microscopy, and X-ray microanalysis have shown that fixation/internalization of quantum dots differ in lymphocytes and neutrophilic granulocytes. Punctate fixation of quantum dots to cell surface is characteristic of lymphocytes, while phagocytosis with perinuclear location of quantum dot aggregations is characteristic of neutrophils. The presence of complex functional groups on the surface of quantum dots can inhibit significantly the fixation/internalization of quantum dots by blood cells.

Characteristics of quantum dots phagocytosis by neutrophil granulocytes.

This article describes the main morphological and biochemical features of neutrophil granulocytes in the process of phagocytosis of two types of quantum dots (CdSe/ZnS-MPA and CdSe/CdSeZnS/ZnS-PTVP). This study is the first to observe the transfer of large aggregates of quantum dots through neutrophils. During this process the cells demonstrate high chemotactic activity. Furthermore, neutrophil pseudopodia underwent alterations during the early stages of phagocytosis. The findings demonstrated that the biochemical profile of neutrophils is practically identical after phagocytosis of both types of quantum dots, but differs significantly from neutrophil metabolism after bacterial phagocytosis.

[Bispecific Antibodies: Formats and Areas of Application].

Bispecific antibodies capable of simultaneously binding two targets have been studied for many years with a view to their implementation in clinical practice. Unique biological and pharmacological properties, as well as the diversity of their formats, make it possible to consider bispecific antibodies as promising agents for use in various procedures: from visualization of intracellular processes to targeted anticancer therapy. Bispecific antibodies help to determine more precisely the therapeutic target, thereby increasing the efficiency of therapy and reducing the probability of side effects. The present review describes the main formats of bispecific antibodies, methods for their generation, and possibilities for practical application.

Microbiota Induces Expression of Tumor Necrosis Factor in Postnatal Mouse Skin.

Tumor necrosis factor (TNF) is a pleiotropic cytokine that regulates many important processes in the body. TNF production in a physiological state supports the structure of lymphoid organs and determines the development of lymphoid cells in hematopoiesis. However, chronic TNF overexpression leads to the development of various autoimmune disorders. Sites of TNF production in the naïve state remain unclear due to the lack of in vivo models. In the present study, we used TNF-2A-Kat reporter mice to monitor the expression of TNF in different tissues. Comparative analysis of tissue fluorescence in TNF-2A-Kat reporter mice and wild type mice revealed constitutive expression of TNF in the skin of naïve adult mice. In the skin of TNF-2A-Kat reporter mouse embryos, no statistically significant differences in the expression of TNF compared to wild type animals were observed. Furthermore, we established that local depletion of microflora with topical antibiotics leads to a reduction in the fluorescence signal. Thus, we assume that the skin microflora is responsible for the expression of TNF in the skin of mice.

Interactions of quantum dots with donor blood erythrocytes in vitro.

The effects of quantum dots CdSe/ZnS-mercaptopropionic acid, (CdSe/CdZnS)ZnS-polyT, and CdSeCdSZnS/polyT/SiO2-NH2 on human erythrocytes were studied. The nanomaterials reduced signifi cantly the erythrocyte sedimentation rate and modified the erythrocyte membrane resistance to induced (acid and hypo-osmotic) hemolysis. Evaluation of the erythrocyte morphology by atomic force microscopy in the control and after exposure to quantum dots showed significant differences in erythrocyte size and changes in their morphology as a result of exposure to the nanomaterials.

[Research of biocompatibility of nanoparticles with fluorescent domen Er/Yb in the neutrophilic granulocytes system].

Fluorescent tags are extensively used for the diagnostic, labeling and marking in vivo and in vitro systems. In this study, fluorescent nanoparticles with Er/Yb lightning center were tested on neutrophilic granulocytes. The main purpose was to idenfity possible toxic effect. The negative impact of fluorophores on the metabolism of neutrophilic and their enzyme systems, on the receptor-mediated cell responses and on the rigidity of cell membranes was shown. The viability of neutrophils (estimated with the use of propidium iodide) after 2 hours of incubation with the fluorescent nanoparticles in concentrations of 10(-4) and 10(-3) mM was 27.0 +/- 6.6 and 19.07 +/- 3.34 %, respectively.

Immunodetection and fluorescent microscopy of transgenically expressed hordeivirus TGBp3 movement protein reveals its association with endoplasmic reticulum elements in close proximity to plasmodesmata.

The subcellular localization of the hydrophobic TGBp3 protein of Poa semilatent virus (PSLV, genus Hordeivirus) was studied in transgenic plants using fluorescent microscopy to detect green fluorescent protein (GFP)-tagged protein and immunodetection with monoclonal antibodies (mAbs) raised against the GFP-based fusion expressed in E. coli. In Western blot analysis, mAbs efficiently recognized the wild-type and GFP-fused PSLV TGBp3 proteins expressed in transgenic Nicotiana benthamiana, but failed to detect TGBp3 in hordeivirus-infected plants. It was found that PSLV TGBp3 and GFP-TGBp3 had a tendency to form large protein complexes of an unknown nature. Fractionation studies revealed that TGBp3 represented an integral membrane protein and probably co-localized with an endoplasmic reticulum-derived domain. Microscopy of epidermal cells in transgenic plants demonstrated that GFP-TGBp3 localized to cell wall-associated punctate bodies, which often formed pairs of opposing discrete structures that co-localized with callose, indicating their association with the plasmodesmata-enriched cell wall fields. After mannitol-induced plasmolysis of the leaf epidermal cells in the transgenic plants, TGBp3 appeared within the cytoplasm and not at cell walls. Although TGBp3-induced bodies were normally static, most of them became motile after plasmolysis and displayed stochastic motion in the cytoplasm.