The Level of Markers Stimulating Growth Factor 2 (ST2), N-Terminal Prohormone of Brain Natriuretic Peptide (NT-proBNP), and D-Dimer in Healthy Volunteer Testers in 21-Day Head-Down -6° Tilt Bed Rest as a Model of Hypodynamia.

The paper presents an analysis of the proteomic composition in relation to both the risk of thrombosis and changes in the state of cardiomyocytes associated with the risk of cardiac fibrosis and heart failure. We examined 12 practically healthy male volunteers exposed to head-down -6° tilt bed rest (HDBR) for 21 days. The revealed decrease in the level of stimulating growth factor 2 (ST2) on days 10 and 21 relative to the initial values (background; 5 days before HDBR) indicated a decrease in the myocardial load and cardiomyocyte extensibility. The level of N-terminal prohormone of brain natriuretic peptide (NT-proBNP) increased on day 2, decreased on days 10 and 21 of HDBR relative to the background levels, and returned to baseline values after the recovery period (5 days after HDBR). The revealed changes in the level of NT-proBNP reflected the increase in circulating blood volume corresponding to HDBR duration and the role of the gravity component in increasing the functional load on the myocardium. Unchanged blood level of D-dimer at all points of the study indicates that there is no risk of thrombosis under the conditions of this study.

[Primary Generalized Glucocorticoid Resistance: a case report].

Primary glucocorticoid resistance (OMIM 615962) is a rare endocrinologic condition caused by resistance of the human glucocorticoid receptor (hGR) to glucocorticoids (GR) and characterised by general or partial insensitivity of target organs to GK. Compensatory activation of hypothalamic-pituitary-andrenal axis results in development of a various pathological conditions caused by overstimulation of adrenal glands. Clinical spectrum may range from asymptomatic cases to severe cases of mineralocorticoid and/or androgen excess. At present time, primary generalized glucocorticoid resistance has been exclusively associated with defects in the NR3C1 gene. Here, we present a case report of an adolescent patient with clinical presentation of glucocorticoid resistance confirmed by detailed endocrinologic evaluation but no confirmed mutations in the NR3C1 gene.

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.

[Advanced glycation end products and oxidative stress as a basis for metabolic abnormalities in patients with type 1 diabetes after successful simultaneous pancreas-kidney transplantation].

AIM: To compare advanced glycation end-products (AGE, RAGE) and 3-nitrotyrosine (3-HT) in patients with DM 1 after successful simultaneous pancreas-kidney transplantation (SPK) and kidney transplantation alone (KTA). To assess relationship between levels of AGE, RAGE, 3-HT and renal transplant (RT) function, carbohydrate and mineral metabolism. MATERIALS AND METHODS: The study included 58 patients who received kidney transplantation in end-stage renal disease (ESRD). 36 patients received SPK. There were performed routine laboratory, examination of AGE, RAGE, 3-NT, parathyroid hormone (PTH), 25(OH)vitamin D, calcium, phosphorus, FGF23, osteoprotegerin (OPG), and fetuin-A levels. RESULTS: All patients after SPK reached normoglycemia (HbA1c 5.7 [5.3; 6.1] %; C-peptide 3.24 [2.29; 4.40] ng/ml) with the achievement of significant difference vs patients after KTA. Arterial hypertension (AH) was more frequent in recipients of SPK before transplantation than after (p=0.008). AH also persisted in greater number of cases in patients after KTA than after SPK. Patients after SPK had higher AGE (р=0.0003) and lower RAGE (р=0.000003) levels. OPG in patients after SPK was significantly higher (р=0.04). The correlation analysis revealed significant positive correlation between 3-HT and OPG (p0.05; r=0.30), RAGE and eGFR (r=-0.52), HbA1c (r=0.48), duration of AH (r=0.34), AGE with HbA1c (r=0.51). CONCLUSION: The results of the "metabolic memory" markers analysis may indicate their contribution to the persistence of the metabolic consequences of CKD and DM 1 after achievement of normoglycemia and renal function restoration and their possible participation in development of recurrent nephropathy, vascular calcification, and bone disorders.

[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.

[Preparation of the dialysis patient with type 1 diabetes mellitus for kidney transplantation].

Kidney transplantation is unique method of renal replacement therapy, allowing to improve quality and duration of life for patients with diabetes mellitus type 1 (DM1) and end-stage renal disease (ESRD) on dialysis therapy. Recently using of innovation technologies for diabetes management and modern immunosuppression enable achieving better results of posttransplant rehabilitation for patients with DM1, especially if kidney transplantation is performed early after initiation of dialysis. Detailed examination of patient with DM1 before potential kidney transplantation is very important to reduce of early and late postoperative complications. Kidney transplantation preparation includes effective glycemic control, adequate dialysis therapy, treatment of diabetes and ESRD complications and concomitant conditions, especially cardiovascular diseases, accounting for kidney transplantation perspective. Furthermore, diagnostics and treatment of any infectious process, timely vaccination, cancer screening are basic approaches of kidney transplantation preparation program, providing the best survival rate of kidney graft and patient.

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.

Syndrome of thyrotoxicosis. Differential diagnosis and treatment.

Since the middle of the twentieth century, there has been a significant change in methods of the diagnosis and treatment of thyroid diseases with thyrotoxicosis syndrome. Previously doctors did not have trouble just with diagnosing diseases that occur with a typical clinical presentation (the Merzeburg triad, a multinodal goiter with fibrillation) because of no possible to determine thyroid hormones. Then in the early 70s years the appearance of immunological methods for estimating hormones in the blood has led to significant changes in our understanding of the variants of thyroid pathology with thyrotoxicosis (TT). Today, the diagnosis of the fact of thyrotoxicosis as a whole is not difficult (except for the confusion of preanalytical errors), but differential diagnosis within the declared syndrome remains extremely relevant to this day. Unfortunately, in the minds of many doctors, these diseases are sometimes perceived as a whole, and in the conditions of the "century of speeds", a modern doctor, extremely limited in time, often unjustifiably prescribes thyreostatic therapy, treatment with radioactive iodine or even surgical intervention after detecting thyrotoxicosis. The old truth "remember that a patient with thyrotoxicosis is a person with a sick heart..." has not lost relevance today. It is very important for the practicing physician be able to navigate in the spectrum of pathologies manifested by the thyrotoxicosis pattern because of the influence of excess thyroid hormones on the cardiovascular system and the hemostasis system. Hereinafter we tried to show diagnostic aspects focusing on differences in pathologies with TT syndrome in a lot of thyroid diseases and even nonthyroid diseases.

Renal experiences of bariatric surgery in patients with type 2 diabetes mellitus.

The review addresses the questions of the literature devoted to the problem of the influence of bariatric surgery on the course of diabetic nephropathy in patients with diabetes mellitus type 2 after achieving a surgically induced remission. This approach was shown to have positive aspects, such as decrease in creatinine, decrease in albuminuria, an increase in GFR, normalization of glycemia and blood pressure, "incretin effect"'s influence on the kidneys. Descriptions of the currently expected pathogenetic mechanisms involved in achieving the observed improvement in microvascular complications of diabetes, namely diabetic kidney disease, are also described in details.

[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.

Label-free study of cosmonaut's urinary proteome changes after long-duration spaceflights.

During the entire time that cosmonauts stay on board the international space station, different extreme space flight factors affect their bodies. In order to find out what physiological changes occur under the influence of spaceflight, different parameters of the human body before and after flights are monitored. Analysis of the urine proteome is one of the most perspective non-invasive methods of condition monitoring. The aim of the study was to perform a comparative semi-quantitative label-free urine proteome analysis of samples collected from 21 cosmonauts before and after long-duration spaceflight at the international space station. For proteomic analysis, urine samples were collected from cosmonauts at three time periods: six months prior to the flight as a background, and on days 1 and 7 of the recovery period after landing. All probes were analyzed by LC-MS/MS, and 256 proteins were identified with more than one unique peptide. The core proteome consists of 50 proteins that are detected in more than 70% of the samples. Label-free semi-quantitative analysis enables us to find 20 proteins which were significantly changed on +1 day and +7 day with respect to background. Most of these proteins participate in the regulation of biological processes, in the regulation of the immune system and in intracellular processes also; some of these proteins are related with stress and response to stimulus. In conclusion, the proteomic analysis of cosmonauts' urine samples provides new data on the human body's adaptation to ground conditions after long-duration spaceflight.

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.