Neutrophil Extracellular Traps (NETs): Opportunities for Targeted Therapy.

Antitumor therapy, including adoptive immunotherapy, inevitably faces powerful counteraction from advanced cancer. If hematological malignancies are currently amenable to therapy with CAR-T lymphocytes (T-cells modified by the chimeric antigen receptor), solid tumors, unfortunately, show a significantly higher degree of resistance to this type of therapy. As recent studies show, the leading role in the escape of solid tumors from the cytotoxic activity of immune cells belongs to the tumor microenvironment (TME). TME consists of several types of cells, including neutrophils, the most numerous cells of the immune system. Recent studies show that the development of the tumor and its ability to metastasize directly affect the extracellular traps of neutrophils (neutrophil extracellular traps, NETs) formed as a result of the response to tumor stimuli. In addition, the nuclear DNA of neutrophils - the main component of NETs - erects a spatial barrier to the interaction of CAR-T with tumor cells. Previous studies have demonstrated the promising potential of deoxyribonuclease I (DNase I) in the destruction of NETs. In this regard, the use of eukaryotic deoxyribonuclease I (DNase I) is promising in the effort to increase the efficiency of CAR-T by reducing the NETs influence in TME. We will examine the role of NETs in TME and the various approaches in the effort to reduce the effect of NETs on a tumor.

[The choice of antibiotic according the results of standard bacteriological analysis and express-method without isolation of pure cultures.]

The efficiency of choosing antibiotics using two methods was analyzed. The first method is based on standard laboratory analysis including isolation of pure cultures of prospective agent and detection of their individual sensitivity to antibiotics. The second method is based on a new express-technique permitting to simultaneously evaluate effect of pharmaceuticals on cultivated and not cultivated yet bacteria without isolation of pure cultures. At that, the second method permits to determine efficiency of antibiotics namely in those concentrations that can be made in the given nidus of infection. The study demonstrated existence of discrepancy between results obtained by using the given methods. The cause of discrepancy can be presence of not cultivated yet bacteria in pathologic sample. In favor of this assumption testify the received data concerning samples from patients where were presented a significant number of spore-former bacteria having additional mechanisms of resistance to antibiotics. Furthermore, the implemented study established antibiotics characterized by the greatest discrepancy of results obtained by applied methods that indicates the necessity of revision of schemes of implementation of start or empiric therapy.

Effects of Multicide, Antibacterial Drug, on Staphylococcus Biomembranes.

Drug penetration into bacterial biomembranes is one of the most important factors determining the efficiency of antibacterial therapy. Multicide, antibacterial drug, is a nanomolecule 1.3-2.0 nm in size, easily penetrating into staphylococcus biomembranes and causing rapid death of bacteria. The drug efficiency depends on its concentration and duration of exposure. Bacteria die as a result of cell wall perforation, which is associated with changes in its morphology and release of DNA from bacterial cell into the environment. Our results indicate the efficiency of primary damage to bacterial wall leading to elimination of biomembranes.

[The choice of antibiotic in microbiological analysis of phlegm.]

The efficiency of application of the test-system "Vy`borAntibiotika" (AntibioticChoice) in incubation of a maximal possible number of bacteria from pathologic material in case of pneumonia was studied. The results of meta-genome analysis permitted to establish that test-system support incubation of practically all bacteria detected in phlegm, including those attributed to so far non-incubated ones. The comparison of the results was carried out concerning a standard detection of sensitivity of bacteria to antibiotics and choice of efficient medicinal according the results of application of test-system "Vy`borAntibiotika". The obtained data demonstrates that test-system permits to choose antibiotic during 6-20 hours without isolation of pure strain.

[The choice of antibiotic in microbiological study of pyoinflammatory processes.]

The efficiency of application of test-system "Antibiotic Choice" was examined concerning evaluation of sensitivity to medications of maximal possible number of bacteria from pathological samples at burn trauma without isolation of pure culture. The results of metagenome analysis demonstrated that test-system permits supporting factually all bacteria discovered in wound discharge, including ones related to not cultivated yet. The comparison was carried out concerning results of standard identification of sensitivity of bacteria to antibiotics and efficient medication according the results of application of test-system "Antibiotic Choice". The obtained results demonstrate that test-system permits choosing antibiotic during 6-20 hours without separation of pure culture.

[CHOOSING ANTIBIOTIC IN MICROBIOLOGICAL ANALYSIS OF INFECTIONS OF URINARY EXCRETION SYSTEM].

The effectiveness of application of test-system "Choice of antibiotic" was evaluated as a tool for incubation of maximal amount of bacteria from pathological material under acute cystitis. The results of meta-genome analysis established that test-system permits supporting growth of practically all bacteria detected in urine, including ones relating to "uncultivated for the present". The comparison of results of standard detection of sensitivity of bacteria to antibiotics and identification of effective pharmaceutical according the results of application of test-system "Choice of antibiotic" as well was implemented It is demonstrated that test- system permits choosing antibiotic during 6-20 hours wiihout isolation of pure strain.

Optimization of the Protocol for the Isolation and Refolding of the Extracellular Domain of HER2 Expressed in Escherichia coli.

Receptor 2 of the human epidermal growth factor (HER2/neu, c-erbB2) is a 185 kDa proto-oncogene protein characterized by an overexpression in some oncological diseases, including 30% of mammary glands cancers, as well as tumors in the ovary, stomach and other organs of the human body. Since HER2- tumor status testing is the essential part of a successful cancer treatment, the expression and purification of substantial amounts of the extracellular domain (ECD) of HER2 is an important task. The production of ECD HER2 in Escherichia coli has several advantages over the use of eukaryotic expression systems, but the bulk of the recombinant product in bacteria accumulates as insoluble protein inclusion bodies. In this study, we obtained ECD HER2 in Escherichia coli as insoluble inclusion bodies and elaborated a simple, efficient, and fast protocol for the solubilization, refolding, and isolation of the protein in soluble form.

Characteristics of bacterial biofilms during long-term culturing.

We studied properties of bacterial biofilms during long-term culturing. The biofilms were preserved for more than 90 days under the chosen conditions. At the same time, amounts of matrix and cells underwent periodic changes. The biomass of the biofilms decreased approximately every 7 days and then returned to the previous values.