Comparative Study of the Levels of IL-1ß, IL-4, IL-8, TNFα, and IFNγ in Stable Course and Exacerbation of Chronic Obstructive Pulmonary Disease of Varying Severity.

In patients with chronic obstructive pulmonary disease, the levels of cytokines IL-1ß, IL-4, IL-8, TNFα, and IFNγ depended on the degree of bronchial obstruction and severity and period of the disease. The maximum levels of IL-4, IL-8, and TNFα were observed in severe chronic obstructive pulmonary disease during exacerbation. The highest concentration of IL-1ß and IFNγ were recorded during activation of inflammation in patients with moderate bronchial obstruction. The revealed correlations between the tested cytokines and spirometry parameters make it possible to consider the levels of these proteins as quantitative markers of systemic inflammation progression.

[Enteroviral (Picornaviridae: Enterovirus) (nonpolio) vaccines].

Non-polio enteroviruses (NPEVs) are ubiquitous and are one of the main causative agents of viral infections in children. NPEVs most commonly infect newborns and young children, due to their lack of antibodies. In children, clinical manifestations can range from acute febrile illness to severe complications that require hospitalization and lead in some cases to disability or death. NPEV infections can have severe consequences, such as polio-like diseases, serous meningitis, meningoencephalitis, myocarditis, etc. The most promising strategy for preventing such diseases is vaccination. No less than 53 types of NPEVs have been found to circulate in Russia. However, of epidemic importance are the causative agents of exanthemic forms of the disease, aseptic meningitis and myocarditis. At the same time, the frequency of NPEV detection in the constituent entities of the Russian Federation is characterized by uneven distribution and seasonal upsurges. The review discusses the epidemic significance of different types of enteroviruses, including those relevant to the Russian Federation, as well as current technologies used to create enterovirus vaccines for the prevention of serious diseases.

[Detection SARS-CoV-2 (Coronaviridae: Coronavirinae: Betacoronavirus: Sarbecovirus) in children with acute intestinal infection in Nizhny Novgorod during 2020-2021].

INTRODUCTION: The novel coronavirus infection COVID-19 is a major public health problem worldwide. Several publications show the presence of gastrointestinal (GI) symptoms (nausea, vomiting, and diarrhea) in addition to respiratory disorders.The aim of this study was the monitoring of RNA of COVID-19 pathogen, coronavirus SARS-CoV-2 (Coronaviridae: Coronavirinae: Betacoronavirus; Sarbecovirus) in children hospitalized with acute intestinal infection (AII), with following molecular-genetic characterization of detected strains. MATERIAL AND METHODS: Fecal samples of children with AII hospitalized in infectious hospital of Nizhny Novgorod (Russia) in the period from 01.07.2020 to 31.10.2021 were used as material for the study. Viral RNA detection was performed by real-time polymerase chain reaction (RT-PCR). The nucleotide sequence of S-protein gene fragment was determined by Sanger sequencing. RESULTS AND DISCUSSION: SARS-CoV-2 genetic material was detected in 45 out of 2476 fecal samples. The maximum number of samples containing RNA of the virus occurred in November 2020 (detection rate of 12.2%). In 20.0% of cases, SARS-CoV-2 RNA was detected in combination with rota-, noro-, and adenoviruses. 28 nucleotide sequences of S-protein gene fragment complementary DNA (cDNA) were determined. Phylogenetic analysis showed that the studied SARS-CoV-2 strains belonged to two variants. Analysis of the S-protein amino acid sequence of the strains studied showed the absence of the N501Y mutation in the 2020 samples, which is a marker for variants with a high epidemic potential, called variants of concern (VOC) according to the World Health Organization (WHO) definition (lines Alpha B.1.1.7, Beta B.1.351, Gamma P.1). Delta line variant B.1.617.2 was identified in two samples isolated in September 2021. CONCLUSION: The detection of SARS-CoV-2 RNA in the fecal samples of children with AII, suggesting that the fecal-oral mechanism of pathogen transmission may exist, determines the necessity to optimize its monitoring and to develop an algorithm of actions with patients with signs of AII under the conditions of a novel coronavirus infection pandemic.

Messenger RNA of FCGR3A and FCGR3B Genes as Monitoring Markers of Clear Cell Renal Adenocarcinoma (a Pilot Study).

The aim of the study was to assess the capabilities of mRNA genes encoding CD16a (FCGR3A) and CD16b (FCGR3B) in tumor samples from patients with renal cancer, and characterize the tumor process in relation to clinical and morphological factors. Materials and Methods: We used 125 tumor samples from patients with a histologically confirmed diagnosis of renal cancer T1-4N0-1M0-1. A method described by Chomczynski and Sacchi was used to isolate nucleic acids. The mRNA levels were determined using a reverse transcription polymerase chain reaction and calculated according to ΔΔCt formula, taking into account the reaction efficiency. Results: mRNA of the FCGR3A gene was detected in all tumor tissue samples under study; in contrast, mRNA of the FCGR3B gene was found only in 92.0% (115/125) of cases. In tumors classified as pT1, the mRNA content of the FCGR3A gene was significantly lower than that in tumor samples of pT3 size. There was the significant increase in the mRNA content of both genes with an increase in tumor grade, as well as in the cases with distant metastases. The presence of a tumor thrombus in the inferior vena cava system was accompanied by a significant increase in the mRNA content of the FCGR3A gene. Conclusion: In tumor tissue samples from patients with clear cell renal cancer, the predominant production of the FCGR3A mRNA was observed in comparison with the FCGR3B mRNA. The revealed relationship of an increased amount of the FCGR3A mRNA and, in some cases, the FCGR3B mRNA with a number of clinical and morphological factors enables to consider the mRNA level of the genes as new monitoring biomarkers.

[Construction of norovirus (Caliciviridae: Norovirus) virus-like particles containing VP1 of the Echovirus 30 (Picornaviridae: Enterovirus: Enterovirus B)].

INTRODUCTION: Enterovirus (nonpolio) infection is widespread all over the world, registered as sporadic cases and large-scale outbreaks and can cause severe lesions such as serous meningitis. Epidemiological studies have shown that enterovirus (Picornaviridae; Enterovirus) variant Echovirus 30 (E30) is the most frequently detected variant in patients with enterovirus meningitis in the Russian Federation. However, no vaccines to prevent the disease caused by this pathogen have been developed so far. One of the promising modern trends in terms of creating vaccine preparations is the use of virus-like particles (VLPs), including chimeric ones containing the biological structures of viruses belonging to different species.The aim of this work was to obtain norovirus (Caliciviridae; Norovirus) VLPs displaying enterovirus Echovirus E30 full-length VP1 on the surface. MATERIAL AND METHODS: The nucleotide sequences of VP1 protein of norovirus genotype GII.4 and VP1 E30 of genotype h circulating in Russia were used. The SN-VP1E30 protein was constructed, in which the shell (S) and the hinge regions of the norovirus VP1 are fused into one molecule with the full-length VP1 of the E30 virus. The protein was expressed in E. coli, purified using affinity chromatography, and characterized by polyacrylamide gel electrophoresis (PAGE) and immunoblotting. VLPs were visualized by electron microscopy. RESULTS: The S N-VP1E30 protein expressed in E. coli as insoluble form, so the conditions for SN-VP1E30 solublisation were defined. Sucrose has been shown to significantly increase the efficiency of renaturation. Electrophoretic mobility comparison of denatured and non-denatured SN-VP1E30 demonstrated that most monomers form high molecular weight compounds. Electron microscopy showed that renatured SN-VP1E30 spontaneously forms empty virus-like particles about 50 nm in diameter. CONCLUSION: Chimeric protein SN-VP1E30 self-assemble into VLPs displaying the VP1 protein of E30 variant that is highly prevalent in Russia. Further immunological research is necessary to characterize VLPs potential for development of the vaccine for enteroviral meningitis prevention.

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.

Influence of Magnetite Nanoparticles and Quantum Dots on the Expression of Reference Genes in Peripheral Blood Cells.

We studied the influence of magnetite nanoparticles (FeO•Fe2O3) and quantum dots (CdSe/ZnS coated with mercaptopropionic acid) on the expression of 5 common reference genes (BA, B2M, PPIA, UBC, and YWHAZ) in peripheral blood cells from 20 volunteers by reverse transcription PCR method. The stability of the expression of reference genes varied depending of the cells type and chemical structure of nanoparticles. The level of YWHAZ mRNA after exposure by nanoparticles demonstrated highest stability in lymphocytes, neutrophils, and monocytes. Stability of YWHAZ expression was confirmed by Western blotting. Our findings suggest that YWHAZ is the most suitable as the reference gene.

[Cancer-testis genes in colon cancer]. / Rakovo-testikuliarnye geny pri rake tolstoi kishki.

The expression of cancer-testis (CT) genes varies with tumor type. There are tumors with high, low, and intermediate gene expressions. Tumor cells of different origin are characterized by ST gene co-expression. The expression of ST genes increases in later stages of tumor development in the presence of metastases. In colon cancer, the tumor samples showed most frequently MAGE-A and SSX mRNA. The peripheral blood samples displayed most commonly XAGE, MAGE-C, and SSX mRNA. In patients with colon cancer, the expression of TSP50, MAGE-A(1-6), and SSX1,2,4 genes was associated with a poor prognosis, that of MAGE-C1 and XAGE1 was related to a favorable prognosis.

[The correlation of expression of MUC1, ICAM1, IL32, FcyR3A and FoxP3 in tumor niduses of patients with breast cancer].

The hypo-glycated mucin I (MUC I) frequently is redundantly expressed in cells of breast carcinoma that is considered as an unfavorable prognostic marker for patients with breast cancer. The purpose of study is to evaluate in tumor areas of patients with breast cancer the interrelationships between expression of MUC I and genes participating in implementation of immune reactions. The technique of polymerase chain reaction with reverse transcription in real-time was applied to investigate expression of genes of MUC I, Fas, ICAMI, FcyR3A, FcyR3B, FoxP3, IL2Ra, IL32 and TNF in tumor areas of 40 patients with breast cancer. The level of mRNA of each of analyzed genes was evaluated relatively to three genes of household. The rate of detection of mRNA MUC I amounted to 80%, mRNA Fas - 95%, mRNA ICAM I - 97.5%, mRNA FcyR3A - 95%, mRNA FcyR3B - 32.5%, mRNA FoxP3 - 85%, mRNA IL2Ra - 87.5%, mRNA IL32 - 75%, mRNA TNF - 95%. The direct correlation was established between levels of mRNA MUC I, ICAM I, IL32, FcyR3A and FoxP3. The most strong relationship was registered between levels of expression of genes MUC I and IL32 (r-0.72; p=0.0001). In comparison with MUC I negative breast carcinoma, MUC I positive breast carcinoma characterized by higher level of mRNA of pro-inflammatory cytokines IL32 and ЕТА, molecules of adhesion ICAM I and molecular marker of T-regulative cells FoxP3. The given profile of expression of genes in tumors of patients with breast cancer demonstrates relationship of expression of MUC I with inflammation and its modulation by T-regulative cells. The effect of expression of analyzed genes on phenotype of cancer cells and course of disease are discussed.

Genetic Instability in Locus rs5498 E469K (A/G) of ICAM-1 Gene in Patients with Colorectal Cancer and Breast Cancer.

Single-nucleotide polymorphism rs5498 E469K (A/G) in ICAM-1 gene modulates functional activity of this protein and is associated with the risk of some diseases. Tumor cell rs5498 E469K (A/G) genotypes were compared with genotypes of patients with colorectal and breast cancer, residents of the Nizhniy Novgorod region. Specimens of peripheral blood and tumor foci from patients with colorectal and breast cancer and peripheral blood specimens from normal subjects were analyzed. The frequency of genotypes of single-nucleotide rs5498 E469K (A/G) sequence in the Nizhny Novgorod population coincide with the frequency of genotypes characteristic of residents of Germany. Comparison of genotypes of peripheral blood and tumor cells from patients with colorectal and breast cancer showed loss of cancer cell heterozygosity. The frequency of heterozygosity loss increases with colorectal cancer progress from one stage to another.

[Role of MUC1 Expression in Tumor Progression].

Mucin 1 (MUC1) is a multistructural and multifunctional protein that is involved in regulating diverse cellular activities. This strongly glycosylated transmembrane protein forms a mucous gel on the surface of epithelial cells that protects the cells from injury. MUC1 acts as a signaling molecule and transcription factor modulating metabolism and resistance to bacterial-induced inflammation. This article presents a review of the relationship between structural and functional changes of the MUC1 and the characteristics of cancer cells. The alteration in MUC1 expression level, a number of structural forms, protein glycosylation and localization occurs in cancer cells. These alterations lead to metabolic reprogramming associated with proliferation, resistance to hypoxia and angiogenesis which affects the survival of cancer cells. Furthermore, cancer cells can take advantage of MUC1 interaction with adhesion molecules for invasion and metastasis. Thus, MUC1 plays a key role both in the homeostasis of epithelial cells and in cancer progression. Understanding the role of MUC1 expression in tumor cells survival is important for the development of new monitoring and therapeutic approaches for the treatment MUC1 positive maligancies.

Levocarnitine normalizes elevated blood level of soluble Fas mRNA in patients with acute myocardial infarction.

Fas-induced apoptosis plays an important role in the mechanisms of tissue injury in myocardial infarction. The level of membrane Fas mRNA was elevated during the postinfarction period in the blood of patients and did not change in response to levocarnitine. The mRNA level of soluble Fas, inhibiting Fas-dependent apoptosis, remained normal during the first 7 days, but increased 14 days after myocardial infarction, which corresponded to previously detected increase of serum level of soluble Fas molecules. Addition of levocarnitine, inhibiting Fas-dependent apoptosis, to therapy caused no changes in the level of soluble Fas mRNA, presumably because of similar effects of soluble Fas and levocarnitine on the apoptotic processes in myocardial infarction.

[Heterogeneous expression of CD38 gene in tumor tissue in patients with colorectal cancer].

The CD38 gene codes a membrane protein which takes part in cell adhesion and catalyzes the formation of cyclic ADP-ribose. Using RT-PCR method we tested the presence of full-size and alternative forms of mRNA CD38 in samples of tumor tissue of patients with colorectal cancer and in tumor cell lines. It was shown that there are the cells in the tumor tissue which expressed CD38 gene. In tumor tissue of patients the alternative form of mRNA CD38 was detected less frequently than full-size form. Cells of lines Colo-205, T-84, HCT15 and HCT116 contained mRNA CD38, in cells of lines Caco-2 and SW-620 mRNA CD38 was absent. In cells of tumor tissue on the first stage of colorectal cancer CD38 gene was expressed in 100% of cases. On the second, third and fourth stages of the disease gene expression was observed less often. The frequency of mRNA CD38 detection not depend on tumor localization, tumor grade and presence of metastases. Using method of restriction analysis CpG methylation was detected in binding sites of transcription factor Sp1 and receptor of retinoic acid (RARE) in all tested samples of tumor tissue independently of the presence or absence of mRNA CD38. The obtained data suggest that in the tumor cells of patients with colorectal cancer the expression of the CD38 gene is heterogeneous.

[Early detection of cancer/testis mRNAs in tumor cells circulating in the peripheral blood of colorectal cancer patients].

Recent studies have suggested that mRNAs transcribed from cancer/testis genes might be used as biomarkers of cancer cells migrating through the bloodstream. Using RT-PCR we evaluated the expression of several cancer/testis mRNAs (MAGEA1-6, GAGE1-8, NY-ESO-1, SSX1, 2, and 4, XAGE1 and MAGEC1) in primary tumors and peripheral blood samples of colorectal cancer patients. The detection rate of at least one of the transcripts was 95% (37/39 samples) for primary tumors and 81% (52/64 samples) for the peripheral blood. Moreover, selected mRNAs were detectable in cellular fraction of the peripheral blood at all stages the disease (14 out of 14 cases), whereas in extracellular fraction of plasma were found only at stages III and IV. Obtained data open a possibility of early diagnosis of colorectal cancer in people at high risk by peripheral blood analysis.

Fluorescence detection of white-beam X-ray absorption anisotropy: towards element-sensitive projections of local atomic structure.

Projections of the atomic structure around Nb atoms in a LiNbO(3) single crystal were obtained from a white-beam X-ray absorption anisotropy (XAA) pattern detected using Nb K fluorescence. This kind of anisotropy results from the interference of X-rays inside a sample and, owing to the short coherence length of a white beam, is visible only at small angles around interatomic directions. Consequently, the main features of the recorded XAA corresponded to distorted real-space projections of dense-packed atomic planes and atomic rows. A quantitative analysis of XAA was carried out using a wavelet transform and allowed well resolved projections of Nb atoms to be obtained up to distances of 10 Å. The signal of nearest O atoms was detected indirectly by a comparison with model calculations. The measurement of white-beam XAA using characteristic radiation indicates the possibility of obtaining element-sensitive projections of the local atomic structure in more complex samples.

Study of DNA interaction with cobalt ferrite nanoparticles.

Interaction of cobalt ferrite nanopowder and nucleic acid was investigated. Superparamagnetic cobalt ferrite nanoparticles (6-12 nm) were prepared by mechanochemical synthesis. Structure of the nanopowder was characterized using X-ray diffraction. It was shown that cobalt ferrite nanoparticles were associated with ssDNA and dsDNA in Tris-buffer resulting in bionanocomposite formation with mass weight relation nanoparticles: DNA 1:(0.083 +/- 0.003) and 1:(0.075 +/- 0.003) respectively. The mechanism of interaction between a DNA and cobalt ferrite nanoparticles was considered basing on the whole set of obtained data: FTIR-spectroscopy, analyzing desorption of DNA from the surface of the particles while changing the chemical content of the medium, and on the modeling interaction of specific biomolecule fragments with surface of a inorganic material. It was supposed that the linkage was based on coordination interaction of the phosphate groups and oxygen atoms heterocyclic bases of DNA with metal ions on the particle surface. These data can be used to design specific magnetic DNA-nanoparticles hybrid structures.

[Genetic diversity of human caliciviruses detected in schoolchildren with gastroenteritis in Nizhni Novgorod].

A total of 2312 children admitted to the Nizhni Novgorod infectious disease hospitals for diagnosed acute gastroenteritis were examined using the RT-PCR. The detection rate of human caliciviruses was found to be 6.5% (range 1.2 to 11.2%). The population of calicivuruses presented with noroviruses genogroups I (16%) and II (81%), and sapoviruses (3%). Noroviruses genogroup II included genotypes GII.2 and GII.4 with a preponderance of the variant CII.4-2006b. The detected sapoviruses were clustered with sapoviruses GI.1, GI.2, and GII.1 by phylogenetic analysis.

[Detection rate of MAGE-A1-A6 mRNA in the peripheral blood of cancer patients].

The mRNA expression of 6 MAGE-A (MAGE-A1-A6) antigens by the tumor focal cells and cancer cells circulating in blood was studied in solid malignant and benign neoplasms. MAGE- A1-A6 mRNA expression was detected in the tumor focal cells in more than 90% of cases of cancer of the breast, lung, and stomach and melanoma cell lines. The detection rate of peripheral blood MAGE-A1-A6-positive tumor cells was 95% in lung cancer, 53% in corpus uteri cancer, 67% in gastric cancer, 63% in breast cancer, 33% in melanoma, and 42% in uterine myoma. MAGE-A3 and MAGE-A4 mRNA expression was more commonly observed.