The microbiome of buried soils demonstrates significant shifts in taxonomic structure and a general trend towards mineral horizons.

Burial mounds represent a challenge for microbiologists. Could ancient buried soils preserve microbiomes as they do archaeological artifacts? To investigate this question, we studied the soil microbiome under a burial mound dating from 2500 years ago in Western Kazakhstan. Two soil profile cuts were established: one under the burial mound and another adjacent to the mound surface steppe soil. Both soils represented the same dark chestnut soil type and had the same horizontal stratification (A, B, C horizons) with slight alterations. DNA samples isolated from all horizons were studied with molecular techniques including qPCR and high throughput sequencing of amplicon libraries of the 16S rRNA gene fragment. The taxonomic structure of the microbiome of the buried horizons demonstrated a deep divergence from ones of the surface, comparable to the variation between different soil types (representatives of the soil types were included in the survey). The cause of this divergence could be attributed to diagenetic processes characterized by the reduction of organic matter content and changes in its structure. Corresponding trends in the microbiome structure are obvious from the beta-diversity pattern: the A and B horizons of the buried soils form one cluster with the C horizons of both buried and surface soil. This trend could generally be designated as 'mineralization'. Statistically significant changes between the buried and surface soils microbiomes were detected in the number of phylogenetic clusters, the biology of which is in the line of diagenesis. The trend of 'mineralization' was also supported by PICRUSt2 functional prediction, demonstrating a higher occurrence of the processes of degradation in the buried microbiome. Our results show a profound shift in the buried microbiome relatively the "surface" microbiome, indicating the deep difference between the original and buried microbiomes.

[Infusion therapy with reamberin in military conditions]. / Ispol'zovanie rastvora reamberina v voennykh usloviyakh.

The article is devoted to infusion therapy with Reamberin 1.5% (Meglumine sodium succinate) in patients with various traumatic injuries. The authors substantiate the relevance and significance of this issue. Specifics and purposes of therapy are considered. The authors reviewed national studies devoted to clinical aspects and determined the main directions of therapy in emergency patients with mechanical injuries, burns, traumatic brain injury and cognitive impairment caused by combat trauma. Moreover, experimental studies of protective properties of Reamberin under combined action of cold, vibration and immobilization, as well as acute massive blood loss are analyzed.

Application of real-time PCR to significantly reduce the time to obtain recombinant MVA virus.

Obtaining a pure recombinant Modified Vaccinia Ankara (MVA) virus is a multistage, time-consuming procedure. We describe a novel single-tube real-time PCR which enables determination of the amount of wild type and recombinant viruses and their ratio in plaques. Use of the real-time PCR significantly reduce the time and efforts needed to obtain purified recombinant MVA. The new approach has been applied to generate recombinant MVAs encoding different SARS-COV-2 antigens.

Hepatitis C virus: The role of N-glycosylation sites of viral genotype 1b proteins for formation of viral particles in insect and mammalian cells.

Hepatitis C virus (HCV) is characterized by considerable genetic variability and, as a consequence, it has 6 genotypes and multitude of subtypes. HCV envelope glycoproteins are involved in the virion formation; the correct folding of these proteins plays the key role in virus infectivity. Glycosylation at certain sites of different genotypes HCV glycoproteins shows substantial differences in functions of the individual glycans (Goffard et al., 2005; Helle et al., 2010) [1], [2]. In this study, differential glycosylation sites of HCV genotype 1b envelope proteins in insect and mammalian cells was demonstrated. We showed that part of glycosylation sites was important for folding of the proteins involved in the formation of viral particles. Point mutations were introduced in the protein N-glycosylation sites of HCV (genotype 1b) and the mutant proteins were analyzed using baculovirus expression system in mammalian and insect cells. Our data showed that, in contrast to HCV 1a and 2a, the folding of HCV 1b envelope proteins E2 (sites N1, N2, N10) and E1 (sites N1, N5) was disrupted, however that did not prevent the formation of virus-like particles (VLP) with misfolded glycoproteins having densities typical for HCV particles containing RNA fragments. Experimental data are supported by mathematical modeling of the structure of E1 mutant variants.

The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells.

The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. The detailed structure of the infectious particle of HCV remains poorly understood. Moreover, the virion assembly and release of virions by the cell are the least understood processes. It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. N-glycans of glycoproteins can influence viral particle formation, virus binding to cell surface, and HCV pathogenesis. We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells.

[EFFICACY OF CLASSICAL CAROTID ENDARTERECTOMY USING TEMPORARY SHUNT].

The study generalized the 15-year experience of classical carotid endarterectomy performance with application of a temporary carotid shunt and an autovenous path in 325 patients. There were analyzed the more frequent complications in standard groups. Advantages of given method were determined and it was specified the ways, which facilitated to reduction of intraoperative risks and increase of technical quality realization of different stages of surgery.

[Role of N-linked glycans in HCV glycoprotein E1 in the folding of structural proteins and formation viral particles].

Envelope proteins of HCV play a major role in virus lifecycle. These proteins are main components of the virion. They are involved in virus assembly. Envelope proteins are modified by N-linked glycosylation which is supposed to play a role in their stability, in the assembly of the functional HCV glycoprotein heterodimer, protein folding and viral entry. The role of N-linked glycosylation sites in HCV E1 protein in structural proteins assembly was analyzed by site-directed mutagenesis in a model system--insect cells producing three viral structural proteins with formation of virus-like particles. Removing of single N-linked glycosylation sites in HCV E1 protein does not affect the efficiency of its expression in insect Sf9 cells. E1 electrophoretic mobility is increasing in parallel with decreasing the number of glycosylation sites. The destroying of glycosylation sites N1 or N5 in E1 influences the assembly of noncovalent glycoprotein heterodimer E1E2--the prototype of natural complex incorporated in virion. The lack of glycans in N1 and N5 sites of E1 was shown to affect the efficiency of its expression in mammalian HEK293 T cells.

[Characterization of hepatitis C virus structural proteins and HCV-like particles produced in recombinant baculovirus infected insect cells].

Three proteins, namely: "core" protein C and glycoproteins E1 and E2, are main structural proteins forming a hepatitis C vius (HCV) virion. The virus structure and assembly, a role of the structural proteins in virion morphogenesis remain unknown because of the lack of an efficient culture system for HCV to be grown in vitro. Using recombinant baculoviruses expressing HCV structural protein genes in insect cells the specific structural proteins at the level of 25-35% relative to a common cell protein content, heterodimers of the glcoproteins, and HCV-like particles have been obtained. It has been demonstrated that recombinant proteins C, E1, and E2 go through the posttranslation modification, the glycoproteins form the non-covalent heterodimer, and HCV-like particles are located in endoplasmatic reticulum membrains of infected cells. An ability of the expressed proteins for forming E1E2 dimers and HCV-like particles was used for studying the role of E1 protein glcosylation upon expression and processing of the glycoproteins.

[Neoangiogenesis and coronary atherosclerosis: diagnostic value of a novel biochemical marker placenta growth factor in patients with ischemic heart disease].

BACKGROUND: Placenta growth factor (PlGF), a member of the vascular endothelial growth factor family, promotes neoarteriogenesis and triggers intraplaque inflammation thereby stimulating atherosclerotic plaque progression and plaque rupture. OBJECTIVE: To investigate prognostic significance of circulating placenta growth factor (PlGF) in coronary artery disease (CAD) patients. METHODS: 78 patients, aged 44-81 years (mean age 61.6+/-13.1 years) with acute myocardial infarction (AMI) (n=19), unstable angina (UA) (n=23), stable effort angina (n=23), and with no evidence of CAD (n=13) were followed-up for at least 48 months. Death, AMI, any revascularization, and hospitalization for UA or progressive effort angina were considered as end points. Plasma levels of PlGF, C-reactive protein (CRP), interleukin-6 (IL-6), neopterin, tumor necrosis factor alpha (TNF-a), haptoglobin and homocysteine were measured at primary admission. RESULTS: During follow up (617+/-263 days) 3 deaths, 1 nonfatal AMI, 4 UA, and 7 angina progression related hospitalizations occurred. Mean event-free survival periods differed significantly between subgroups of patients with low (<7.5 pg/ml), medium (7.5-20.5 pg/ml), and high (>20.5 pg/ml) PlGF levels (1038+/-56, 729+/-55, and 578+/-63 days, respectively). Logrank survival in patients with low PlGF was significantly better than in high PlGF subgroup (p=0.038). PlGF levels did not correlate with age, lipid levels, blood pressure and smoking. A significant positive correlation was found between PlGF and haptoglobin (r=0.34, p=0.028), homocysteine (r=0.455, p=0.044), neopterin (r=0.31, p=0.048), and carotid intima-media thickness. CONCLUSION: Elevated PlGF plasma levels predict worse prognosis in CAD patients; PlGF levels correlate with haptoglobin, neopterin, and homocysteine blood levels and with the carotid artery intima-media thickness.

[Vasculopathy of the grafted heart: the synergism of proinflammatory and proatherogenous factors and viral infection].

The authors analyze the pathogenetic significance of hyperhomocysteinemia, antiphospholipid syndrome, the hyperexpression of cell adhesion molecules, inflammation, and oxidative disorders for, as well as the role of viral infections in the development of coronary artery disease of the grafted heart. The paper shows that viral infections in recipients lead to the development of proinflammatory, proatherogenous, and prothrombogenous status, expressing themselves in an increase in the corresponding laboratory markers in recipients' blood plasma, and points out the role of viral infection in the pathogenesis of coronary artery disease of the transplanted heart. Control and treatment of viral infections, as well as pharmacocorrection of proinflammatory, proatherogenous, and prothrombogenous status would made it possible to influence the development of coronary artery disease of the grafted heart.

[Inflammation and apoptosis in relation with the effectiveness of bone marrow cell transplantation to patients with chronic heart failure].

The authors analyze the pathogenetic significance of inflammation, immune activation, and apoptosis in the development of heart failure and the functional regeneration of left ventricular myocardium after treatment including autologic bone marrow cell transplantation. The paper shows that autologic bone marrow cell transplantation influences the same mechanisms that cause heart failure, namely inflammation, apoptosis, and neurohumoral mechanisms. Investigations have made it possible to formulate guidelines for the prediction and monitoring of the effects of treatment including autologic bone marrow cell transplantation in patients with heart failure.

[Prognostic value of C-reactive protein and neopterin in autologous bone marrow cells transplantation].

Relationship between levels of C-reactive protein (CRP) and neopterin and changes of left ventricular function was studied in 23 patients with heart failure (mean age 48.5+/-11.5 years) subjected to transplantation of autologous bone marrow cells (TABMC). Cells were administered intracoronary irrespective of coronary revascularization (in 11 patients) or during angioplasty (in 5 patients), intracoronary or intra-cardially during aorto coronary bypass surgery (in 7 patients). Reverse correlation was observed between left ventricular ejection fraction (EF) and stroke volume (SV) and blood plasma levels of CRP and neopterin. Efficacy of functional regeneration of the myocardium assessed 1 month after TABMC was related to blood plasma levels of CRP and neopterin before treatment. In all patients with normal initial levels of markers of inflammation EF and SV rose. Among patients with initially elevated levels of CRP and neopterin more positive changes of EF and SV occurred in those subjects in whom lowering of inflammation markers took place during 1 month after TABMC.