Influence of a Passive Tilt Test on the Proteomic Composition of the Blood of Healthy Humans.

In order to identify changes in the blood proteome of healthy volunteers after passive tilt test carried out on day 19 of head-down bed rest, a chromato-mass-spectrometric analysis of samples of dried blood spots was carried out. It was revealed that the body's response to the tilt test was characterized by a decrease in the level of HDL and kininogen-1. After the tilt test, we observed an increase in the level of vimentin, vitamin K-dependent protein C, Wnt signaling pathway proteins, proteins involved in autophagy and adaptive immune response, focal adhesion proteins, vascular damage marker S100A8, PEDF regulator, and some proteins of the heart: cardiac actin ACTC1 and transcription factor GATA4. The obtained results lay the foundation for future research in the framework of identifying the risks of developing cardiovascular changes in astronauts after space flights.

Characteristics of blood plasma proteome changes associated with the hemorrhagic purpura of cosmonauts on the first day after long-term space missions.

The interest in the role of the gravitational factor during landing after long-term space flights (SF) leads to the search for various innovative approaches to assessing the compliance of external changes observed by clinicians. The results of special research methods such as Omics technologies that may reflect physiological responses to the conditions created during landing are of great interest. Our purpose is to compare the blood plasma proteome changes associated with the trauma and endothelial dysfunction processes prior to launch and on the day of landing, as well as the groups of cosmonauts with and without the secondary hemorrhagic purpura. In our study, the concentrations of 125 plasma proteins in 18 Russian cosmonauts, measured using targeted proteomic analysis based on liquid chromatography and tandem mass spectrometry were analyzed. The results reveal the trends of 12 proteins participating in the processes that trigger hemorrhagic purpura under the effect of re-entry g-forces. Exposure to intense g-forces and return to the gravity are the key factors for external manifestations of changes in the body systems induced by a long-term stay in space microgravity. Our results may be useful for further research to experts in gravitational physiology, aviation and space medicine.

Relationship of collagen as the component of the extracellular matrix with the mechanisms of autonomic regulation of the cardiovascular system under simulated conditions of long-term isolation.

The study of proteins - potential markers, associated signal transduction pathways, and their targets - provides a new understanding of the fundamental mechanisms occurring at the level of regulatory processes in the cardiovascular system (CVS), especially in space flight, as well as in model experiments that reproduce its individual effects on the human body. The article presents the results of studies in an experiment with 120-day isolation within the framework of the SIRIUS project in which 6 volunteers aged 28 to 44 years (three men and three women) participated. SIRIUS (Scientific International Research in Unique Terrestrial Station) is the international research project, which studies the issues of biomedical and psychological support of long-term manned space flights. The possible involvement of collagen different types, an extracellular matrix protein, in the mechanisms of autonomic regulation of the CVS was studied. Using chromatic mass spectrometry in urine samples and analysis of heart rate variability, we have established that the extracellular matrix collagen, which is present, in particular, in the structure of the blood vessel wall, are markers associated with the modulating effect of the autonomic nervous system on the regulatory mechanisms of blood circulation. We hypothesized that these proteins may be a biomarker of the autonomic balance in the regulatory mechanisms of the circulatory system. In addition, these proteins can also be markers of the aging process, which increases the risks of developing autonomic dysfunction of the cardiovascular system (dominance of sympathicotonia) and changes in the quality of the tissue of the heart muscle and blood vessels, provoking the development of prenosological conditions and diseases of the cardiovascular system.

[Age features of heart rate variability in cosmonauts according to urine proteomics data.]

As the life of the International Space Station (ISS) is lengthening, Russian crews are dominated by cosmonauts over 40 years of age who have repeatedly made a space flight (SF) and have a long stay on the ISS. In order to study the age-related features of the regulation of heart rate variability (HRV) in cosmonauts, based on proteomics data, we analyzed simultaneously the obtained data on the proteomic composition of urine samples and electrocardiogram (ECG) parameters, followed by an analysis of heart rate variability (HRV) in cosmonauts in the age group of 46,5±3,4 (n=12). The survey was carried out in the period of preparation for the next flight. The urine proteome was studied by mass spectrometry and analyzed by bioinformatic methods. Heart rate variability was assessed by software methods for analyzing ECG fragments. For the first time, data based on proteomics on the age-related features of the regulation of heart rate variability in cosmonauts are presented. According to age periodization, cosmonauts are in the middle age of the second period. The concentrations of mucin 1, type VI collagen, and cadherin 13 were shown to change with age. A decrease in HRV was also observed with increasing age. There was an increase in sympathetic influence and a progressive decrease in parasympathetic influence.

[Characteristics of age-dependent changes in the urine´s proteom composition of the of the healthy persons (experimental and theoretical study).]

For the first time proteins are described, reliably increasing and decreasing in urine with age in the range of 20 to 60 years. The combinations of proteins associated with changes in immune processes, violation of blood reology, including the risk of coagulopathy, anticancer defense mechanisms, insulin signaling pathway, changes in cell characteristics are characterized division and quality of the newly formed fabric. Thus, the age dynamics of the main processes triggers a cascade of reactions manifested in the closure of «pathological biochemical circles¼ that form the prerequisites for the development of diseases and, over time, clinical manifestations.

[Chromato-mass spectrometric analysis of urine proteins associated with the functions of Toll-receptors in a healthy person under conditions of 17-day isolation.] / Хромато-масс-спектрометрический анализ белков мочи, ÑÐ¾Ð¿Ñ€ÑÐ¶ÐµÐ½Ð½Ñ‹Ñ Ñ функциями Toll-рецепторов у здорового человека в ÑƒÑÐ»Ð¾Ð²Ð¸ÑÑ 17-суточной изоляции.

Under controlled conditions of 17-day isolation (Sirius-17 experiment), the protein composition of urine was studied in 6 healthy test volunteers-3 women and 3 men. Collection of samples in the form of a second freely separated morning urine fraction was carried out in the background (seven days before the experiment), as well as 1 day after the end of exposure. Chromatographic-mass-spectrometric semi-quantitative analysis of the protein composition of samples was performed on a system consisting of an Agilent 1100 chromatograph and an LTQ-FT Ultra hybrid mass spectrometer using bioinformatics resources UniProtKB, GeneOntology. An asymptomatic change in the immune defense system of kidney tissue after isolation in a closed hermetic object is associated with a change in the content of 7 proteins that provide functional activity of the TLR tubules of the kidneys - FcRIII, MUC1, Galectin-3, Ficolin-2, APOA1, FLNA, FCGR3A and Clusterin. These proteins are found to be useful biomarkers in the study of physiology and kidney diseases. They can be attributed to candidates for protein markers of the initial stages of impaired recognition by the epithelium of renal tubules of bacteria with known pathogenic potential.

Secretome of Cultured Human Endothelial Cells in Simulated Microgravity.

The secretome of human umbilical vein endothelial cells (HUVEC) cultured under static conditions and in modeled microgravity for 24 h was studied by chromatography-mass spectrometry. In the secretome of cells exposed to microgravity, we identified a group of microtubule proteins including many structural elements of microtubules and regulatory proteins interacting with Rho-GTPases. Hence, reorganization of actin cytoskeleton and microtubules induced by microgravity is under complex regulation mediated by Rho proteins.

[Features of the proteome of the urine in chronic pyelonephritis.]

Clinical observation and examination of 12 patients with chronic pyelonephritis (CPN) were performed. The first group (GI) included patients with exacerbation of the disease. In the comparison group (GII)- the same patients after 1.5-3 months after completion of treatment, without clinical manifestations of exacerbation of CPN. Laboratory signs of acute renal damage were not revealed in all examined patients. Additionally, urine was collected in the afternoon after Breakfast, in the form of a freely separated 2nd fraction and its sample preparation, consisting of the stages: recovery, alkylation, protein deposition and proteolysis using trypsin. The resulting polypeptide mixture was separated by liquid chromatography in three repetitions and analyzed on a system consisting of Agilent 1100 chromatograph and ltq-FT ultra hybrid mass spectrometer. A list of proteins was obtained, indicating the number of peptides by which they were identified, and the parameters of its reliability. Most of the information about the obtained proteins was obtained from UniProt databases. Identified and analyzed 10 proteins that differ significantly in occurrence in the clinical group of patients in the period of exacerbation of PN. The appearance of these proteins in urine in 1patients with exacerbation of chronic PH allows us to consider them as potential biomarkers directly associated with inflammation and damage to the epithelial lining of the renal tubules.

[Analysis of Urine Proteome in Patients With Postinfarction Cardiosclerosis Combined With Hypertensive Disease for Assessing Endothelial Dysfunction].

In our study urine protein composition of 18 healthy volunteers was compared with that of 18 patients with ischemic heart disease and concomitant hypertension. Liquid chromatography-mass-spectrometry (LC-MS) analysis of the second fraction of morning urine was carried out using nano-line high performance liquid chromatograph and hybrid mass spectrometer. The analysis revealed 23 proteins expressed in the endothelium, according to the information contained in the database Bgee, and 49 proteins, with direct functional link with the processes in the endothelium in the reconstruction of associative networks using ANDSystem program. Comparison of urine proteome of healthy people and patients with postinfarction cardiosclerosis revealed proteins specific for patients with cardiovascular disease. Thus, proteins vitronectin, syndecan-4, a histidine rich glycoprotein, endothelial protein C receptor, colony stimulating factor, cathepsin D and sekretogranin-1 may be considered as potential markers for cardiovascular diseases. Further research in this area should be conducted for clinical and experimental verification of these hypotheses.

Effect of Different Levels of Salt Consumption on Urine Protein Composition during 105-Day Isolation Analyzed with the Use of opoSOM Software.

We investigated changes in the urine protein composition of healthy volunteers in controlled conditions during 105-day isolation (Mars-500 program) at different levels of salt consumption. We used newest proteomic techniques based on chromatography-mass spectrometry and various methods of bioinformatics including opoSOM. The period of observation can be divided into three intervals with different dynamics of protein excretion: early (week 1-6), intermediate (week 7-11) and late interval (week 12-15). We identified about 10 different groups of co-detected proteins, which are directly affected by periods with different-levels of salt consumption. We also determined the biological functions of these proteins, tissue specificity and signaling pathways that involve them.

PROTEOMIC ANALYSIS OF URINE UNDER CONTROLLED SALT INTAKE IN PROJECT <>.

Specifics of urine proteome is sensitive to a multitude of factors. One is nutrition or entrance in organism of main nutrients including salt (NaCI). Purpose of the investigation was to study the proteomic composition of healthy human urine in the controlled environment of a 105-day isolation experiment (project <>) with various levels of salt intake. Analysis was performed using the present-day proteomics techniques based on chromatography-mass spectrometry and bio-informatics options. An attempt was made to correlate changes in processes and physiological systems with the controlled salt intake. As a result, a list of proteins directly responsible for different salt intake during the experiment and then a list of tissues where these proteins express predominantly were compiled; besides, analysis of the processes these proteins are involved in was performed.