Influence of a 1000 Times Weakened Magnetic Field on Embryogenesis and Ontogenesis of the Japanese Quail in Several Generations.

The paper presents experimental data on the influence of a 1000-fold weakening of the Earth's magnetic field on the embryonic and postembryonic development of the Japanese quail in three generations. It has been shown that the weakening of the earth's magnetic field by a factor of 1000 affects the formation of blood vessels in Japanese quail embryos, in particular, causing a decrease in angiogenesis in seven-day-old embryos of both the first generation (F1) and the next two ones (F2 and F3). Pathological and anatomical studies of embryos of different ages in three generations have revealed various pathologies associated with vascular system disorders, as well as disorders in the development of the beak and eyes. In the ontogenesis of F3 quails, there is a decrease in the hatchability of chicks.

The Biological Threat: The Threat of Planetary Quarantine Failure as a Result of Outer Space Exploration by Humans.

The paper presents the results of experiments with spore-forming bacteria and microscopic fungi performed in the framework of the Russian Research Program outside the International Space Station. It has been found that microorganisms not only survive in this extreme environment, but also retain reproductive ability. Moreover, most microorganisms exhibit an increase in biochemical activity and resistance to antimicrobial agents, specifically antibiotics. These findings are of obvious interest to the developers of both planetary quarantine methods and biomedical safety systems for manned space exploration missions. In addition, they demonstrate the necessity of experiments on the exposure of bio-objects to simulated environmental factors beyond Earth's magnetosphere.

Effect of microgravity on glial cell line-derived neurotrophic factor and cerebral dopamine neurotrophic factor gene expression in the mouse brain.

UNLABELLED: Mice were exposed to 1 month of space flight on the Russian biosatellite BION-M1 to determine its effect on the expression of genes involved in the maintenance of the mouse brain dopamine system. The current article focuses on the genes encoding glial cell line-derived neurotrophic factor (GDNF) and cerebral dopamine neurotrophic factor (CDNF). Space flight reduced expression of the GDNF gene in the striatum and hypothalamus but increased it in the frontal cortex and raphe nuclei area. At the same time, actual space flight reduced expression of the gene encoding CDNF in the substantia nigra but increased it in the raphe nuclei area. To separate the effects of space flight from environmental stress contribution, we analyzed expression of the investigated genes in mice housed for 1 month on Earth in the same shuttle cabins that were used for space flight and in mice of the vivarium control group. Shuttle cabin housing failed to alter the expression of the GDNF and CDNF genes in the brain structures investigated. Thus, actual long-term space flight produced dysregulation in genetic control of GDNF and CDNF genes. These changes may be related to downregulation of the dopamine system after space flight, which we have shown earlier. © 2015 Wiley Periodicals, Inc. SIGNIFICANCE: Our results provide the first evidence of microgravity effects on expression of the GDNF and CDNF neurotrophic factor genes. A considerable decrease in mRNA level of GDNF and CDNF in the nigrostriatal dopamine system was found. Because both GDNF and CDNF play a significant role in maintenance and survival of brain dopaminergic neurons, we can assume that this dysregulation in genetic control of GDNF and CDNF genes in substantia nigra could be among the reasons for the deleterious effects of space flight on the dopamine system.

Effect of actual long-term spaceflight on BDNF, TrkB, p75, BAX and BCL-XL genes expression in mouse brain regions.

Mice of C57BL/6J strain were exposed to 1-month spaceflight on Russian biosatellite Bion-M1 to determine the effect of long-term actual spaceflight on the expression of genes involved in the processes of neurogenesis and apoptosis. Specifically, we focused on the genes encoding proapoptotic factor BAX, antiapoptotic factor BCL-XL, brain-derived neurotrophic factor (BDNF) and BDNF receptors TrkB and p75. Spaceflight reduced the expression of the antiapoptotic BCL-XL gene in the striatum and hypothalamus, but increased it in the hippocampus. To estimate environmental stress contribution into spaceflight effects we analyzed spaceflight-responsive genes in mice housed for 1 month on Earth in the same shuttle cabins that were used for spaceflight, and in mice of the laboratory control group. It was shown that 1-month shuttle cabin housing decreased BCL-XL gene expression in the striatum but failed to alter BCL-XL mRNA levels in the hippocampus or hypothalamus. Spaceflight failed to alter the expression of the proapoptotic BAX gene in all investigated brain structures, although the insignificant increase of the BAX mRNA level in the hippocampus of spaceflight mice was found. At the same time, shuttle cabin housing produced insignificant decrease in BAX gene expression in the hippocampus. In contrast to the BCL-XL gene, genes encoding BAX, BDNF as well as TrkB and p75 receptors did not respond to 30-day spaceflight. Thus, long-term spaceflight (1) did not affect the expression of genes encoding BDNF as well as TrkB and p75 receptors, (2) produced dysregulation in genetic control of the neuronal apoptosis, (3) implicated BCL-XL as the risk factor for spaceflight-induced behavioral abnormalities.

[Development of paste-type food for experiments with mice onboard unmanned spacecrafts].

One of the crucial issues of handling animals in space flight is availability of food and water supply systems. However, water supply for animal experiments onboard unmanned spacecrafts is a particularly formidable problem. To maintain laboratory mice in space flight, a paste-type food was formulated on the basis of the standard extruded combined food for laboratory rodents (PK-120) with a 18.3% wet protein content and moisture raised to 68%. Food manufacturing technology for rodents reproduces essentially the one for quails that had shown good advantages in an experiment with adult birds aboard orbital complex Mir. The proposed food was tested with white laboratory mice (males and females) of mature 37-d age. According to the data of blood clinical analysis and visceral organs morphology investigations, feeding with the paste-type food without additional water over 21 days did not change the life weight of mice or food digestibility and availability. These biological test results gave go to feed mice in the Bion-M1 and synchronous ground experiments with the paste-type food.

[The BION-M1 project: overview and first results].

Biosatellite BION-M1 was launched on April 19 and landed on May 19, 2013. The mission program was largely a continuation of the earlier flown 11 BION projects, FOTON-M2 and FOTON-M3. The biosatellite was inhabited by a great variety of living organisms used for experiments and studies in gravitational physiology, gravitational biology, biotechnology, astrobiology and radiation biology, dosimetry and spectrometry. This was the first time in the history of national biology and physiology when male mice C57bl/6 were chosen for a long-term space experiment focused upon molecular biology investigations. Unfortunately, because of technical failures during the flight a part of the animals were lost. However, the major objectives were attained through reconsideration of biomaterial division among investigators and completion of virtually the total scope of investigations.

[Experimental studies with mice on the program of the biosatellite BION-M1 mission].

Purpose of the BION-M1 project was laying the evidence and technological basis for addressing the medical issues of future remote space exploration missions by humans. The program of researches with the use of mice was focused on elicitation of cellular and molecular mechanisms of the muscular, cardiovascular and immune reactions to extended exposure in microgravity. The comprehensive studies combined lifetime measurements with investigations of mice tissues and cells by dint of the cutting-edge morphological, biochemical and molecular biology techniques. Males of mice C57/BL6 aged 4 to 5 months were chosen as the object of studies. They were distributed into the flight, ground control and two vivarium (laboratory control) groups and investigated immediately on return and after 7 days of readaptation. Some of the physiological functions were recorded throughout the flight. To ensure wellbeing of the animals in the experiments and to enhance data quality, prior to launch the mice were specially trained so as to accommodate to the group living, eating space food, and in-flight stress factors. Those of the mice that were designated for lifetime investigations were tested and received training pre-launch.

[Space experiments on the development of biological systems for the human life].

Over the past 22 years, the Institute of Biomedical Problems has stubbornly continued the investigations with higher plants aimed at the development of cultivation technologies suitable for the conditions of space flight. Analysis of the results of 24 plant experiments performed aboard orbital complex MIR and the ISS Russian segment evidenced the ability of higher plants to grow, develop and reproduce inside spacecraft living compartments. Space crops were normal as compared with the laboratory controls. Microbial contamination of the plants was within the normal limits; no pathogen has been detected on plant surfaces. Plants did not change genetically, at least in four space generations. It should be noted that the presence of greenhouse on board the ISS also has a marked positive effect on wellbeing of people living in the close environment and isolation from Earth's biosphere. In the context of the above, the higher plants might become a secure and beneficial part of the life support system for crews on space exploration missions.

[Histogenesis of Japanese quail bone and cartilage tissues at the final stages of embryonic development in microgravity].

The article reports the results of comparative histological studies of skeleton development in 14- and 16-day Japanese quail embryos grown in space flight and in the 1-g gravity. The investigation revealed retardation of cartilage replacement by bone in the femur and tibia in space embryos as compared with their controls. Perichondral ossification metaphysis was reached by day 14 both in the space and control embryos. Destruction of cartilaginous diaphysis advanced onto the proximal and distal parts including the periphery. Tibia and femur cartilaginous cores in space embryos were destructed worse than in the controls in consequence of insufficient minerals supply. Perichondral ossification in the 16-day space and control embryos was alike close to completion spreading as far as the epiphysis. Long bones metaphysis was abundant in monomorphic cells as a growth bank existing during and 7 days post hatching. However, absence, in contrast to the controls, of osteogensis sites in long bones epiphysis, suggested retardation of chondrocytes calcification in these areas, as well as of ossification in space embryos.

[Spinal marrow development in Japanese quail embryos and chicklings in the spaceflight environment].

Results of studying spinal marrow histogenesis in age varying Japanese quail embryos and chicklings whose embryonic development took place in microgravity are presented. Histological comparison of spinal cords showed retardation of morphogenetic development, lumber spine especially, in the group of flight embryos manifested as incomplete proliferative activity during migration processes and nervous cell differentiation. Like on earth, in space microgravity chicklings also hatched on day 17 of embryogenesis. Multiform marrow changes in space chicklings were a nervous cells reaction to the functional state of organism in the micro-g environment.

[Early bone and cartilage histogenesis in embryonic Japanese quails in the conditions of microgravity].

The article presents the results of a comparative histological investigation of skeletal bones genesis in Japanese quail embryos developed in the spaceflight microgravity (space group) and laboratory (control group). Total preparations of 4-day-old embryos from both groups demonstrated clearly that the cartilaginous anlage of the femoral bone had central, dyaphisial, 2 epiphysial and 2 proliferation zones. By day 7 of embryogenesis, cartilaginous anlages had grown in size in both groups due to intensive chondrocytes multiplication and gain in the intercellular substance mass. Tibial cuff in space embryos measured half and femoral cuff was 2.3 times smaller in comparison with these parameters in the control group. In addition, intensity of chondrocyte multiplication was reduced Histological profiles of the femur and tibia in 10-day old embryos of the control pointed to enhancement of osteogenesis. The metaphysis zone contained distinct mitosis figures on different stages of division. Bone deposition could be seen below the peristoma. The osteogenesis cuff spread up to the femoral anlage metaphysis; cartilage was calcined. Space embryos display retard osteogenesis. There were ingrown blood vessels in the region of cartilage destruction; however, vessels grown in the periosteum were less in number as compared with the laboratory control. Also, the perichondral ossification layer was considerably thinner, whereas the osseous cuff was 1.3 and 1.45 times shorter in the femur and tibia, respectively. To sum up, the histological investigation of bones from 4-, 7- and 10- day old Japanese quail embryos demonstrated retardation of osteogenesis in the conditions of microgravity.

[Histogenesis of the liver of Japanese quail embryos developed in the conditions of microgravity].

The article reports the results of histogenetic studies of the liver extracted from Japanese quail embryos and hatchlings that had passed the embryonic and postnatal development in microgravity. Comparison of the liver from the flight and laboratory embryos showed identity of organogenesis and histogenesis processes in space and on Earth. However, it should be noted that early histogenetic development of the space embryo liver had been retarded which was concluded from an enlarged lumen in bile capillaries and loose arrangement of epithelium girders as compared with the control On day-10 post flight, the flight embryos exhibited liver induration through narrowing of sinusoids lumen. Haemopoietic zones became less numerous. By the end of embryonic development these histological differences of the flight and control liver disappeared essentially. Organogenesis and histogenesis of the liver obtained from the hatchlings that had spent 5 days in microgravity did not depart from the norm. However, inability of the birds to adapt to microgravity and ensuing starvation was the reason for dystrophic changes in the liver, venous congestion and increased histoleucocytic activity. All the liver changes faded away soon alter the hatchlings were settled in favorable life conditions

[Dynamics of calcium utilization for skeleton formation in Japanese quail embryos under the microgravity condition].

Tibia and femur osteogenesis was studied in embryonic lower limbs developed in microgravity. The maximal difference in ash content of shell in the flight and control groups was registered on days 4, 10 and 14 amounting to 10.27; 9.56 and 12.95% respectively. Shell analysis for calcium showed the largest difference between the groups on day 4 (8.94 mg). However, this difference was not seen already on days 14 and 16. Hence, according to the results of the investigation, although shell calcium utilization and osteogenesis in flight embryos of the Japanese quail were retarded, by the time of hatching the morphological and chemical parameters of the lower limb bones were essentially same as of the ground controls.

[Stress response genes expression analysis of barley Hordeum vulgare under space flight environment].

Transcriptome of barley Hordeum vulgare grown aboard International Space Station (ISS) was analyzed by means of microarray. It was revealed 500 genes with mRNA level, changed more than two folds in space environment. Among them are genes encoding stress response proteins, videlicet Heat Shock Proteins (HSP), Pathogenesis-Related Proteins (PR) and Antioxidant Proteins. Further analysis of these genes by real time PCR showed enhanced transcription level of Reactive oxygen Species (ROS) scavenging genes. The mRNA level of superoxide dismutase (sod) was 6 folds higher in space environment when compare to Earth conditions. Glutamyl transferase gene expression was enhanced 24 times in space. Transcription of catalase gene (cat) was increased 18 times and of ascorbate peroxidase was increased 3 times in space in comparison with ground control. For the first time it was shown that space flight environment may induce oxidative stress in plants.

[Effect of thermal shock on seed resistance].

The work was performed in order to evaluate possible negative effect of high temperatures on biological properties of seeds in space experiment BIORISK-MSN-2. Laboratory experiments showed that seeds possess high enough resistance to extended exposure to high temperatures. Results of studying effects of positive temperatures in the region of 40 to 100 degrees C on biological properties of seeds of 19 species and varieties of higher plants belonging to three different families lead to the conclusion that heat stability of seeds depends on temperature value, period of exposure, species, variety and individual peculiarities of plants. These data can be useful in future interpretation of results of experiment BIORISK-MSN-2.

[Histogeny of the visceral organs of embryonic Japanese quails developed in the micro-g environment].

The paper reports the results of organogenic and histogenic investigations of the visceral organs of embryonic Japanese quails incubated in the microgravity aboard orbital complex Mir. Investigations of the gastrointestinal tract failed to reveal macrodeviations in the organs' structure, differentiation of layers or cells along the full length of the entodermal canal. However, comparison with the ground controls exhibited poor development of stromal connective tissues in the flight embryos evidencing loose arrangement and small number of fibers. Local hyperplasia in the duodenal epithelium was due to the proliferation rather than differentiation processes; it could affect food intake and parietal digestion in the flight chicks. Though the Japanese quail embryos developed in space microgravity had some deviations, their digestic apparatus was mature to uptake and assimilate food.

[Biological systems of life support for space crews: some results and anticipations].

One of the challenges for space biology and medicine is resolution of lots of problems of biomedical support of humans in the extreme environment of space flight. These problems include also designing of robust and efficiently functioning life support systems (LSS). The paper gives an overview of the investigations of ground-based BLSS with human subjects conducted in Russia and other countries. Also, it contains the basic data of studying the BLSS photoautotrophic components (higher plants) in the series of experiments with the total duration of 630 days fulfilled on orbital complex Mir and the series of experiments with the total duration of 820 days in the ISS Russian segment. Analysis of the results from the BLSS investigations on Earth and in space flights drives to the conclusion that some of the BLSS components, greenhouses specifically, can be integrated even now into the currently used systems of space crew life support.

[Effect of hypoxic argon-containing gas mixtures on developing organism].

Experimental data on the impact of argon-containing hypoxic breathing mixtures on the early embryonic growth of Japanese quail (Coturnix coturnix japonica dom.) are presented. The results showed that after 4-day incubation of eggs in an oxygen-nitrogen breathing mixture (13.8-15.0% O2, 86.2-85.0% N2) only 16.7% embryos reached the stage of 3.5-d development. Introduction in the hypoxic mixture of argon (55% of the total volume) increased the number of embryos on the stage of 3.5-4 day of normal morphological development to 60%. It became evident from the analysis that embryos developed in the O2-Ar-N2 hypoxic mixture had 25% less pathologies as compared with embryos developed in a largely nitrogen-containing gas mixture. Hence, we received the experimental evidence that 55% of argon in a hypoxic breathing mixture with up to 15% of oxygen is favorable to the early embryonic growth of Japanese quail stimulating metabolic processes in germ organism.

[The study of the genetic effects in generation of pea plants cultivated during the whole cycle of ontogenesis on the board of RS ISS].

Results of studies on growth and development of offspring of two genetically marked dwarf pea lines planted during the whole ontogenesis cycle in the Lada space greenhouse on board of Russian Segment of International Space Station (RS ISS) are presented. The offspring of M1 and M2 plants grown from seeds formed during space flight was examined under conditions of Earth-based. Cultivation. It had been shown that growth and developmental characteristics, frequency of chromosome aberrations in primary root meristem and level of molecular polymorphism revealed in individual plants via RAPD method show no significant differences between offspring of "space-grown" and control seeds.