Quantitative target proteomics of chromosome 13 human blood plasma proteins.

Quantitative proteomic analysis of 50 blood plasma samples of healthy volunteers who underwent a comprehensive medical examination and were found eligible for space flights was performed. As a result of directed mass spectrometric analysis, signals for 128 proteins, which accounted for nearly 40% of the total number of chromosome 13 gene products, were detected. The analysis of interindividual variation of concentrations of chromosome 13 proteins showed the presence of a pool comprising 41 proteins with a low variation (CV < 30%), which can potentially be used as biomarkers.

[The Status of Hemostasis System in Hypoxic Nitrogen-Oxygen and Argon-Oxygen Diving Gases].

In this study the effect of factors of hermetic chamber with modified gas medium on the hemostasis system is analyzed in order to estimate and to compare different diving breathing gases. The parameters characterizing pro-, anticoagulant as well as fibrinolytic components of hemostasis were determined using clotting, chromogenic and immunological methods. The applied exposure did not affect the activity and regulatory potential of hemostasis significantly; however, the nitrogen-oxygen and argon-oxygen diving gases have a different effect on the hemostasis functioning, especially in the recovery period.

[COMPREHENSIVE ASSESSMENT OF THE HEMOSTASIS SYSTEM IN TEST-SUBJECTS IN EXPERIMENT "MARS-500"].

The paper is devoted to the hemostasis studies in the course of long-term (520 d) isolation in an airtight chamber. Measured parameters included activated partial thromboplastin time (APTT), international normalized ratio (INR), thrombin time (TT); concentrations of fibrinogen (FBG), plasminogen (PG), Willebrand factor (WF), tissue factor pathway inhibitor (TFPI), and tissue plasminogen activator (TPA), thrombomodulin (TM); activities of the coagulation cascade factors (II, V, VII, X, VIII, IX, XI, XII), antithrombin III (ATIII), protein C (PC), C1-inhibitor (C1), α2-antiplasmin (AP), TPA and TFPI. The investigation revealed a diversity of changes in the plasma fibrinogen concentration, slower blood coagulation in the intrinsic pathway and the final stage, and a relative rise in the activities of ATIII and PC-inhibited factors. The rest parameters showed different trends.

[INVESTIGATION OF IRON, ZINC, STABLE STRONTIUM AND LITHIUM CONTENT IN BIOLOGICAL FLUIDS AND TISSUES DURING EXPERIMENTAL SPACE FLIGHT SIMULATION].

In an experiment with space flight simulation, atomic emission spectral analysis with inductively coupled argon plasma was employed to measure iron, zinc, stable strontium and lithium in blood serum and its ultrafiltered fraction, and excretion with daily urine and hair. Monitoring of serum iron and its ultraviolet fraction showed good balance of these parameters in all periods of the experiment. Blood serum was found to contain exclusively iron bound with protein carriers. In the experiment, serum zinc was distinctively dependent on the nutrient status. Stable strontium excretion with daily urine can be a biological indicator of its homeostasis. The experiment factors had no effect on the lithium form in serum, i.e. lithium was invariably present in the ionized form and in quantities equal to ultrafiltered lithium in all blood samples in all periods of the experiment.

Functional changes in neutrophils and psychoneuroendocrine responses during 105 days of confinement.

The innate immune system as one key element of immunity and a prerequisite for an adequate host defense is of emerging interest in space research to ensure crew health and thus mission success. In ground-based studies, spaceflight-associated specifics such as confinement caused altered immune functions paralleled by changes in stress hormone levels. In this study, six men were confined for 105 days to a space module of ~500 m(3) mimicking conditions of a long-term space mission. Psychic stress was surveyed by different questionnaires. Blood, saliva, and urine samples were taken before, during, and after confinement to determine quantitative and qualitative immune responses by analyzing enumerative assays and quantifying microbicide and phagocytic functions. Additionally, expression and shedding of L-selectin (CD62L) on granulocytes and different plasma cytokine levels were measured. Cortisol and catecholamine levels were analyzed in saliva and urine. Psychic stress or an activation of the psychoneuroendocrine system could not be testified. White blood cell counts were not significantly altered, but innate immune functions showed increased cytotoxic and reduced microbicide capabilities. Furthermore, a significantly enhanced shedding of CD62L might be a hint at increased migratory capabilities. However, this was observed in the absence of any acute inflammatory state, and no rise in plasma cytokine levels was detected. In summary, confinement for 105 days caused changes in innate immune functions. Whether these changes result from an alert immune state in preparation for further immune challenges or from a normal adaptive process during confinement remains to be clarified in future research.

520-d Isolation and confinement simulating a flight to Mars reveals heightened immune responses and alterations of leukocyte phenotype.

During interplanetary exploration, chronic stress caused by long term isolation and confinement in the spacecraft is one of the major concerns of physical and psychological health of space travelers. And for human on Earth, more and more people live in an isolated condition, which has become a common social problem in modern western society. Collective evidences have indicated prolonged chronic stress could bring big influence to human immune function, which may lead to a variety of health problems. However, to what extent long-term isolation can affect the immune system still remains largely unknow. A simulated 520-d Mars mission provided an extraordinary chance to study the effect of prolonged isolation. Six healthy males participated in this mission and their active neuroendocrine and immune conditions were studied with saliva and blood samples from all participants on chosen time points during the isolation period. As a typical neuroendocrine parameter, stress hormone cortisol was measured in the morning saliva samples. Immune phenotype changes were monitored through peripheral leukocyte phenotype analysis. Using an ex vivo viral infection simulation assay we assessed the immune response changes characterized by the ability to produce representative endogenous pro-inflammatory cytokines. The results of this study revealed elevated cortisol levels, increased lymphocyte amount and heightened immune responses, suggesting that prolonged isolation acting as chronic stressors are able to trigger leukocyte phenotype changes and poorly controlled immune responses.

[Status of the osteoclast-activating system in cosmonauts after long-duration missions to the International Space Station].

The results of studying the system of osteoprotegerin/ receptor activator of nuclear factor kappa-B ligand (OPG/RANKL) in 22 cosmonauts after long-duration (124 to 199 days) ISS missions are presented. Immediately on return to 1 g, changes were observed in OPG and RANKL serum levels and the ability to produce unstimulated and stimulated PGA of peripheral blood mononuclear cells in vitro. Individual variability of these changes was noticed. Our findings suggest that the cytokine OPG/RANKL-system is involved in bone remodeling in members of long-duration space missions.

[Effect of physical countermeasures against support load deficiency on the hemostasis system in an experiment with 7-day immersion].

In an experiment with 7-d dry immersion for testing the mechanic foot and electrical muscle stimulation procedures of compensating the support load deficiency the following biochemical parameters were studied: activated partial thromboplastin time, prothrombin time, prothrombin index, international normalized ratio, thrombin time (TT), fibrinogen, soluble fibrin-monomer complexes, D-dimer (DD), plasminogen (PG); antithrombin III activity (ATIII), protein C (PC) and alpha2-antiplasmin (AP). Control subjects (immersed w/o stimulation) did not exhibit significant shifts in the hemostasis system: however, they decreased ATIII, PC and PG in the post-experimental period. Mechanic foot stimulation did not have a noteworthy effect on the hemostasis system functioning during immersion, whereas electromyostimulation increased procoagulant and fibrinolytic activity in blood plasma.

[Status of the water-soluble component of the antioxidant defense system in the conditions of 520-day isolation].

In the 520-d chamber experiment within the international project Mars-500 blood samples of 6 male test-subjects of 28 to 39 years of age were analyzed for water-soluble antioxidants: total bilirubin and uric acid; in addition, total antioxidant capacity of blood plasma was determined. Maximal values of these parameters were associated with the most stressful periods of the experiment, i.e. adaptation to the life in isolation and confinement, simulation of the egress onto Martian surface, and change of the diet. On attainment of the homeostatic equilibrium the parameters stabilized on levels slightly lower relative to baseline (pre-isolation) values. Therefore, dynamics of the water-soluble antioxidants reflected adequately the homeostatic reactions to and compensation by organism of the effects of the 520-day life in isolation and confinement.

[Immunological aspects of a piloted mission to Mars].

The paper deals with the results of the effects of 520-day isolation and confinement modeling some elements of a mission to Mars on the immune system. Longitudinal analyses revealed that the mechanisms of adaptive response of the human immune system to the conditions of extremely long isolation led to a change of the parameters, characterizing innate and adaptive immunity. Among them the most important are: changes in the signaling PRRs--TLR, manifested in the reduction of the percentage of circulating monocytes and granulocytes expressed on its own surfaces TLR2, TLR4 and TLR6, decreases early NK cell activation potential, increases in the percentage T- and B-lymphocytes, that expressed early activation marker CD69 after adequate stimulation, and in production of cytokines in response to PHA stimulation. The active mobilization of the mechanisms of adaptive immunity, the implementation of the function of the level of immunity to a qualitatively different level, apparently, should be taken as a sign of adaptive adjustment of an organism in response to the complex influence of unfavorable factors, aimed at the preservation of immune homeostasis.

[Reference values of Russian cosmonauts blood biochemical indices].

Within the period of 2000 to 2012 the values of 40 diagnostically significant biochemical indices in venous blood were being determined during the pre-flight clinical and physiological examination of 28 Russian cosmonauts aged 35 to 54, members of main and back-up crews of expeditions to the International Space Station. The above examination was conducted 45-30 days prior to the launch. Taking account of the fact that each of the most of the cosmonauts performed several flights over the mentioned period and they were repeatedly included in back-up crews over and over again, each of cosmonauts participated in pre-flight examinations 1 to 5 times. Reference values were calculated for each of the studied indices. It was found that reference interval boundaries for 15 indices were narrower as compared to the generally accepted ranges. The upper boundaries of activity reference ranges of a number of enzymes, as well as of concentration of some metabolites of energy and plasticity metabolism were raised relative to mean population values. Thus, it was found that Russian cosmonauts being a specific professional team featured inherent reference values which differed from mean population values in a number of blood biochemical indices. It is conditioned by peculiarities of selection, physical training and psychic and emotional status of crew members during a pre-flight period.

[Investigation of the copper content in blood serum and its ultrafiltrate in the conditions of experimental space flight simulation].

In experiment MARS-500 on remote space flight simulation, atomic emission spectral analysis with inductively coupled argon plasma was used to measure serum copper and its ultrafiltration fraction. It was shown that, in contrast to baseline data, the Cu serum level varied within the normal physiological boundaries throughout the entire simulation experiment. These variations were sync in all the serum samples. In several periods in the experiment reductions in serum Cu within the baseline range were equivalent to simultaneous reductions in ceruloplasmin. Amount of the free Cu (ultrafiltration) fraction, i.e. not bound to proteins in blood serum, was constant at all times. Since the simulation experiment is not a perfect analogy of space flight because of nonreproducibility of some factors, ionizing radiation specifically, we should take into consideration shifts in the serum copper balance caused by the action of space radiation on human organism. Radiation-induced imbalance is not only a marker of disturbed oxidase function of blood but also points to a mechanism of potential pathological outcome of toxic excessive copper accumulated in the brain, liver and kidney.

[Intensity of lipid peroxidation and antioxidant protection system indices in the course of a 3-day "dry" immersion experiment].

In the course of a 3-day "dry" immersion experiment involving participation of five male test volunteers aged 25 to 40, in the blood serum of participants were determined the content of LPO (lipid peroxidation) products, namely diene conjugates, malon dialdehyde, shiffbases, as well as the values of the antioxidant protection system (AOP) indices - the concentration of tocopherol as the main lipid antioxidant and the level of general antioxidant activity. During the immersion action no deviations of indices under study from the background values were revealed, with the exception of a certain increase in tocopherol concentration within two hours after experiment beginning. At the restoring period after two hours upon immersion completion a reliable increase of lipid peroxidation products, particularly diene conjugates in participants' blood serum was made evident. At this, the tocopherol content was significantly below the background values. It is therefore concluded that the test participants' adaptation to the simulated weightlessness conditions shows no pronounced stress component, whereas bringing back to normal vital functions after exposure to immersion occurs with significantly pronounced stress reaction development, as demonstrated by considerable growth of LPO processes against the background of AOP system functional activity reducing.

[Magnesium level in human organism during 105-day isolation].

Total and ionized magnesium in blood serum and in daily urine were determined before (baseline values) and on days 30, 60 and 105 of the experiment with 105-d isolation and confinement (Mars-105)/ Magnesium in hair was investigated before (baseline values) and on day-105 of the experiment. The investigations were performed using atomic emission spectrometry with inductively coupled argon plasma. Changes in magnesium were most significant over the initial 30 days in the experiment. Reduction of serum magnesium was accounted for by the fall in the ionized fraction. In organism magnesium is controlled by the ion-regulatory function of the kidney and varies with individuals. Levels of ionized magnesium in blood serum and excreted with daily urine can serve as indicators of stress resistivity .

[Metabolic effects of physical countermeasures against deficient weight-bearing in an experiment with 7-day immersion].

Metabolic effects of physical countermeasures against deficient weight-loading were studied in three groups of 21-30 y.o. volunteers for 7-d dry immersion. Blood serum was investigated for 38 biochemical parameters that characterize myocardium, skeletal musculature, hepatobiliary system, kidney, pancreas, GI tract, prostate, and protein-nucleic, carbohydrate, electrolyte and mineral metabolism. Seven-day DI w/o countermeasures (n = 5) increased concentration of conjugated bilirubin, suppressed activities of muscular (creatine phosphokinase MM) and myocardial enzymes (CPK MB, OBDH), and caused an upward trend in cholesterol, its atherogenic LDP fraction and triglycerides. Mechanic sole stimulation (n = 6) intensified, within the physiological norm, erythrocyte hematolysis raising total bilirubin and potassium. Despite the stimulation, activity of muscle and myocardial enzymes made a decrease. Blood creatinine decreased to a less extent than in the immersed group w/o stimulation, however, lipid parameters did not rise. High-frequency stimulation of the lower leg and hip muscles in the course of immersion (n = 5) was noted to heighten the activity of muscle enzymes and potassium level in blood beyond the physiological norm. Change in creatinine did not reach a statistical significance and lipid metabolism parameters were not different from baseline values. Application of these physical methods of counteracting deficiency of weight bearing did not interfere with redistribution of body liquids due to immersion. Values of the parameters under study were mostly within the normal limits throughout the experimental exposure suggesting absence of pathological developments during DI or in consequence of physical stimulation. Therefore, the reactions were obviously of normal adaptive character.

[Dynamics of blood concentration of neurospecific proteins and risk of neuropathy development in the conditions of 105-day confinement].

Six male volunteers (aged 25 to 40 years) were subjects in all-round psychophysiological, hormonal and immunological studies before, in and after 105-day isolation and confinement. Blood was drawn and the 16-factorial Cattell personality inventory was filled out every 30 days. Concentrations of blood hormones, neurospecific proteins and cytokines point to a close interrelation between antibody titers to myelin-associated glycoprotein and changes in the parameters of metabolism and reproduction-related hormones, as well as cytokines and individual psychophysiology (extra-introversion, dominance, intropunitiveness, social contact selectivity, etc.), and suggest a minimum risk of demyelinizing neuropathy due to exposure to the conditions of isolation and confinement.

[Congenital human immunity during 5-day dry immersion].

The system of congenital immunity was studied in 12 essentially healthy males 18 to 26 years of age subjected to 5-day dry immersion without use of countermeasures. Peripheral blood was analyzed for monocytes, granulocytes and lymphocytes expressing the TLR2+, TLR4+, TLR6+, CD11b+, CD14+, CD16+, CD18+, CD24+, CD36+, CD54+, CD56+ and CD206+ receptors. Expression of early activation marker CD69 on lymphocytes-natural killers was studied in unstimulated and interleukin-2 activated mononuclear cell cultures. The negative shifts in the congenital immunity system in some volunteers at the end of immersion and during recovery can be considered as warnings about depletion of the system reserve and increase of the risk of infectious diseases such as caused by normal microflora which typically does not provoke pathological reactions of the host.

[Dynamics of the body composition, neurohumoral and psychophysiological status of humans in the conditions of 105-day isolation and confinement].

Before, in and after the experiment with 105-day isolation and confinement, 6 male volunteers from 25 to 40 years of age rationed NaCl and performed integral impedancimetric, psychological and hormonal investigations. Every 30 days blood collection for hormonal measurements was combined with filling of Cattell's 16 personal factor questionnaire. Parameters of total body fluid, body mass, basic exchange, specific hydration and basic exchange were determined. The results showed that the experimental conditions did not affect significantly body composition, metabolism or neurohumoral regulation; the metabolic variations were largely associated with motivation for and value orientation in accommodation, to the permissible extent, of the controlled diet and work/rest schedule to personal needs. In addition, it was found that evolution of the psychophysiological status of humans in isolation and confinement is governed primnarily by personality characteristics and, to a less degree, specifics and length of exposure to the artificial environment; thus, in the opinion of the volunteers normoxic, normobaric and slightly hypercapnic (0.15-0.65% CO2) atmosphere was comfortable and harmless to health. Analysis of the whole data array verified the expressed interrelation of neuroendocrine and psychophysiological parameters as well as shifts in body basic exchange and mass, salt intake and hydration rate in the conditions of isolation and confinement.

[Interrelation of dynamics of blood lipids and bone mineral density in humans during 370-day bed rest].

Comparison of bone mineral density and fatty-acid blood content in 9 human subjects exposed to 370-d bed rest revealed correlation of the loss in femoral neck density with parameters of lipid exchange. On day-46 of BR, the absolute lipids content in erythrocyte membranes and blood serum decreased considerably (1.5-2 times) when compared with baseline data. At the end of the experiment, lipids content in serum, on the contrary, surpassed baseline values 2-3 times: however, it remained lowered in erythrocyte membranes of the control group till day-280 of BR. Arachidonic acid correlated with prostaglandins PGE2 and PGF2alpha involved in regulation of osteoclasts and osteoblasts activities. Correlation of decreases in femoral neck density and unsaturated fatty acids in blood serum and erythrocyte membranes varied with the human subjects.

[Intensity of lipid peroxidation and antioxidant protection system indices during re-adaptation period after long-term space flights at the international space station].

In the blood serum of seventeen members of crews which participated in 14 orbital expeditions to the International Space Station with the duration of 125 to 217 days, during the pre-flight period and on the day of landing on the 1st, 7th and 14th days of the rehabilitation period (RP) the content of LPO products was determined, namely diene conjugates (DC), malon dialdehyde (MDA), shiffbases (SB) and the main lipid oxidant - tocopherol (TP). The group of astronauts who made landing in the Space Shuttle spacecraft (8 persons) and the group of astronauts who accomplished space mission in the Soyus TM spacecraft (9 persons) demonstrated a decrease in DC and MDA levels with a rise in TF concentration in the course of the rehabilitation period. Changes in the group of the American spacecraft astronauts were more pronounced. LPO inhibition during the rehabilitation period is recognized [treated] as an adequate reaction to the stress caused by re-adaptation to the ground conditions. Also are discussed probable mechanisms of intergroup differencies in LPO intensity degree: biomembrane phase state changing under the influence of overloads during de-orbiting and stress response intensity during landing in different types of spacecraft.