Differentiation of cartilaginous anlagen in entire embryonic mouse limbs cultured in a rotating bioreactor.

Mechanisms involved in development of the embryonic limb have remained the same throughout eons of genetic and environmental evolution under Earth gravity (1 g). During the spaceflight era it has been of interest to explore the ancient theory that form of the skeleton develops in response to gravity, and that changes in gravitational forces can change the developmental pattern of the limb. This has been shown in vivo and in vitro, allowing the hypergravity of centrifugation and microgravity of space to be used as tools to increase our knowledge of limb development. In recapitulations of spaceflight experiments, premetatarsals were cultured in suspension in a bioreactor, and found to be shorter and less differentiated than those cultured in standard culture dishes. This study only measured length of the metatarsals, and did not account for possible changes due to the skeletal elements having a more in vivo 3D shape while in suspension vs. flattened tissues compressed by their own weight. A culture system with an outcome closer to in vivo and that supports growth of younger limb buds than traditional systems will allow studies of early Hox gene expression, and contribute to the understanding of very early stages of development. The purpose of the current experiment was to determine if entire limb buds could be cultured in the bioreactor, and to compare the growth and differentiation with that of culturing in a culture dish system. Fore and hind limbs from E11-E13 ICR mouse embryos were cultured for six days, either in the bioreactor or in center-well organ culture dishes, fixed, and embedded for histology. E13 specimens grown in culture dishes were flat, while bioreactor culture specimens had a more in vivo-like 3D limb shape. Sections showed excellent cartilage differentiation in both culture systems, with more cell maturation, and hypertrophy in the specimens cultured in the bioreactor. Younger limb buds fused together during culture, so an additional set of E11.5 limb buds was cultured with and without encapsulation in alginate prior to culturing in the bioreactor. Encapsulated limbs grown in the bioreactor did not fuse together, but developed only the more proximal elements while limbs grown in culture dishes formed proximal and distal elements. Alginate encapsulation may have reduced oxygenation to the progress zone of the developing limb bud resulting in lack of development of the more distal elements. These results show that the bioreactor supports growth and differentiation of skeletal elements in entire E13 limb buds, and that a method to culture younger limb buds without fusing together needs to be developed if any morphometric analysis is to be performed.     

Teratogenic effects of gravitational changes.

In investigating the effect of gravitational changes on development, it is instructive to think of altered gravity (delta g) as a teratogen--that is, an environmental factor influencing development. Observed effects on skeletal development include: suppression of morphogenesis in centrifuged mouse limb buds; advanced fusion stages in centrifuged mouse palates; smaller crown rump lengths (CRL) and decreased number of pregnancies in centrifuged rats and mice; altered differentiation of growth plates in young growing rats in space; and decreased length of calcified long bone regions in fetal rats exposed to microgravity in utero. These studies show that delta g is able to alter development in vivo and in vitro and suggest that delta g operates, at least in part, at the cellular level.     

Exposure to altered gravity affects all stages of endochondral cartilage differentiation.

Chondrogenesis has a number of well-defined steps: (1) condensation, which involves cell aggregation, adhesion and communication; (2) activation of cartilage genes, which is accompanied by rounding up of the cells and intracellular differentiation; and (3) production and secretion of cartilage specific matrix molecules. Our studies show that each of these steps is affected by exposure to gravitational changes. Clinorotation and centrifugation affected initial aggregation and condensation. In the CELLS experiment, where cells were exposed to microgravity after some condensation occurred preflight, intracellular differentiation and matrix production were delayed relative to controls. Once cartilage has developed, in rats, further differentiation (hypertrophy, matrix production) was also affected by spaceflight and hind limb suspension. For the process of chondrogenesis to proceed as we know it, loading and other factors present at 1g are required at each step of the process. This requirement means that not only will skeletal development and bone healing, processes involving chondrogenesis, be altered by long term exposure to microgravity, but that continuous intervention will be necessary to correct any defects produced by altered gravity environments.     

Do heavy ions cause microlesions in cell membranes?

Heavy ions are a hazard in manned deep space missions. It has been theoretically postulated that when they interact with cells, localized damage in the forms of "microlesions" may occur. Purported morphological evidence of these lesions, however, has not been confirmed in the most extensively studied tissue so far, the cornea. Recent morphological evidence from rat corneas demonstrated that holes in membranes do not form as consequence of heavy ion irradiation. This does not mean, however, that some other form of damage is excluded. For example such damage may be physiological in nature, impairing the ability of cells or tissues to function properly. In order to uncover any physiological effects, we investigated the microlesion question by monitoring the electrical potential difference across the endothelium of rat corneas in vitro before, during, and after irradiation. When the corneas were exposed to 1 Gy of 56Fe ions (450 and 600 MeV/a.m.u.), we detected no effect on this parameter. These results suggest that direct physical damage to cell membranes, as predicted by the microlesion theory, does not take place.     

空间科学任务协同设计论证平台

空间科学任务协同设计论证存在设计方案耦合强,数据一致性差,数据变更难,数据与流程脱节等问题.为了解决上述问题,建立了典型空间科学任务的多层数字化模型;采用图形化方式对论证流程进行建模,并建立流程与数据的映射关系;通过共享数据池的方式为多岗位用户提供多方案数据协同机制;采用消息总线对数据变更进行及时提醒;利用方案依赖关系矩阵来判别方案耦合关系,最终自动合并任务总体设计方案.作为一个分布式平台,空间科学任务协同设计平台采用Eclipse RCP和Spring技术架构,整合了Hibernate、工作流Activiti5等中间件,提供统一门户,支持多岗位、多任务、多方案、多版本的管理能力,提供论证流程监控、数据协同交互等功能.结合某空间科学任务论证验证了该平台的有效性.      

Chondrogenesis in aggregates of embryonic limb cells grown in a rotating wall vessel.

Previous studies in this lab have shown that chondrogenesis is affected in growth plates of rats exposed to microgravity, and in micromass cultures of embryonic limb mesenchyme differentiating in space. In order to provide a three dimensional aspect not seen in the micromass system, and a tissue homogeneity not possible with explants of limb or limb elements, and to alleviate certain difficulties regarding crew time and stowage, we began culturing embryonic limb cells in Rotating Wall Vessels (RWV). First, these cells were attached to beads, and grown for up to 65 days in a type of RWV known as STLV at the Johnson Space Center. During this time, the cells and beads aggregated and the aggregates continued to increase in size, and differentiated into Alcian blue staining chondrocytes. Because our intent was to use these aggregates for implanting into bony defects in addition to their use in studies of chondrogenic regulation at 1g and microgravity, aggregates of these cells without beads were grown in the commercially available version of the STLV, and their ability to ossify when subcutaneously implanted assessed.     

Changes in the central nervous system during long-duration space flight: implications for neuro-imaging.

The purpose of this paper is to review the potential functional and morphological effects of long duration space flight on the human central nervous system (CNS) and how current neuroimaging techniques may be utilized to study these effects. It must be determined if there will be any detrimental changes to the CNS from long term exposure to the space environment if human beings are to plan interplanetary missions or establish permanent space habitats. Research to date has focused primarily on the short term changes in the CNS as the result of space flight. The space environment has many factors such as weightlessness, electromagnetic fields, and radiation, that may impact upon the function and structure of the CNS. CNS changes known to occur during and after long term space flight include neurovestibular disturbances, cephalic fluid shifts, alterations in sensory perception, changes in proprioception, psychological disturbances, and cognitive changes. Animal studies have shown altered plasticity of the neural cytoarchitecture, decreased neuronal metabolism in the hypothalamus, and changes in neurotransmitter concentrations. Recent progress in the ability to study brain morphology, cerebral metabolism, and neurochemistry in vivo in the human brain would provide ample opportunity to investigate many of the changes that occur in the CNS as a result of space flight. These methods include positron emission tomography (PET), single photon emission computed tomography (SPECT), and magnetic resonance imaging (MRI).     

Biological dosimetry in Russian and Italian astronauts.

Large uncertainties are associated with estimates of equivalent dose and cancer risk for crews of long-term space missions. Biological dosimetry in astronauts is emerging as a useful technique to compare predictions based on quality factors and risk coefficients with actual measurements of biological damage in-flight. In the present study, chromosomal aberrations were analyzed in one Italian and eight Russian cosmonauts following missions of different duration on the MIR and the international space station (ISS). We used the technique of fluorescence in situ hybridization (FISH) to visualize translocations in chromosomes 1 and 2. In some cases, an increase in chromosome damage was observed after flight, but no correlation could be found between chromosome damage and flight history, in terms of number of flights at the time of sampling, duration in space and extra-vehicular activity. Blood samples from one of the cosmonauts were exposed in vitro to 6 MeV X-rays both before and after the flight. An enhancement in radiosensitivity induced by the spaceflight was observed.     

Development of countermeasures for medical problems encountered in space flight.

By the turn of this century, long-duration space missions, either in low Earth orbit or for got early planetary missions, will become commonplace. From the physiological standpoint, exposure to the weightless environment results in changes in body function, some of which are adaptive in nature and some of which can be life threatening. Important issues such as environmental health, radiation protection, physical deconditioning, and bone and muscle loss are of concern to life scientists and mission designers. Physical conditioning techniques such as exercise are not sufficient to protect future space travellers. A review of past experience with piloted missions has shown that gradual breakdown in bone and muscle tissue, together with fluid losses, despite a vigorous exercise regimen can ultimately lead to increased evidence of renal stones, musculoskeletal injuries, and bone fractures. Biological effects of radiation can, over long periods of time increase the risk of cancer development. Today, a vigorous program of study on the means to provide a complex exercise regimen to the antigravity muscles and skeleton is under study. Additional evaluation of artificial gravity as a mechanism to counteract bone and muscle deconditioning and cardiovascular asthenia is under study. New radiation methods are being developed. This paper will deal with the results of these studies.     

Global approach to simulation: a gateway to long-term human presence in space.

The establishment of an autonomous European manned space capability is an objective set up by the ESA Council Meeting at the ministerial level, in 1985/1987. ESA's Long-Term Programme Office (LTPO), charged of the preparation of the programme for a European Manned Space Infrastructure (EMSI), started during 1988 to build up an intellectual framework in the domain of long-duration manned space missions. EMSI scope was eventually extended to embrace Moon/Mars missions and bases. Several exploratory studies on problems related to human factors in long-duration space missions were initiated by LTPO. The work of an ad-hoc group of experts (SIMIS Group) has been focused during 1989/1990 on the planning for simulation of such missions with a broad mandate, covering the physiological, psychological and operational aspects of long-duration exposure to microgravity and isolation/confinement. Preliminary results of SIMIS activities are reported. The HYDREMSI experiment, carried out in a terrestrial, analogous environment for 72 days during 1989, is described as an example of the envisaged simulations.     

Materials trade study for lunar/gateway missions.

The National Aeronautics and Space Administration (NASA) administrator has identified protection from radiation hazards as one of the two biggest problems of the agency with respect to human deep space missions. The intensity and strength of cosmic radiation in deep space makes this a 'must solve' problem for space missions. The Moon and two Earth-Moon Lagrange points near Moon are being proposed as hubs for deep space missions. The focus of this study is to identify approaches to protecting astronauts and habitats from adverse effects from space radiation both for single missions and multiple missions for career astronauts to these destinations. As the great cost of added radiation shielding is a potential limiting factor in deep space missions, reduction of mass, without compromising safety, is of paramount importance. The choice of material and selection of the crew profile play major roles in design and mission operations. Material trade studies in shield design over multi-segmented missions involving multiple work and living areas in the transport and duty phase of space mission's to two Earth-Moon co-linear Lagrange points (L1) between Earth and the Moon and (L2) on back side of the moon as seen from Earth, and to the Moon have been studied. It is found that, for single missions, current state-of-the-art knowledge of material provides adequate shielding. On the other hand, the choice of shield material is absolutely critical for career astronauts and revolutionary materials need to be developed for these missions. This study also provides a guide to the effectiveness of multifunctional materials in preparation for more detailed geometry studies in progress.     

Neuroplasticity changes during space flight.

Neuroplasticity refers to the ability of neurons to alter some functional property in response to alterations in input. Most of the inputs received by the brain and thus the neurons are coming from the overall sensory system. The lack of gravity during space flight or even the reduction of gravity during the planned Mars missions are and will change these inputs. The often observed "loop swimming" of some aquatic species is under discussion to be based on sensory input changes as well as the observed motion sickness of astronauts and cosmonauts. Several reports are published regarding these changes being based on alterations of general neurophysiological parameters. In this paper a summing-up of recent results obtained in the last years during space flight missions will be presented. Beside data obtained from astronauts and cosmonauts, main focus of this paper will be on animal model system data.     

Operations of a spaceflight experiment to investigate plant tropisms

Plants will be an important component in bioregenerative systems for long-term missions to the Moon and Mars. Since gravity is reduced both on the Moon and Mars, studies that identify the basic mechanisms of plant growth and development in altered gravity are required to ensure successful plant production on these space colonization missions. To address these issues, we have developed a project on the International Space Station (ISS) to study the interaction between gravitropism and phototropism in Arabidopsis thaliana. These experiments were termed TROPI (for tropisms) and were performed on the European Modular Cultivation System (EMCS) in 2006. In this paper, we provide an operational summary of TROPI and preliminary results on studies of tropistic curvature of seedlings grown in space. Seed germination in TROPI was lower compared to previous space experiments, and this was likely due to extended storage in hardware for up to 8 months. Video downlinks provided an important quality check on the automated experimental time line that also was monitored with telemetry. Good quality images of seedlings were obtained, but the use of analog video tapes resulted in delays in image processing and analysis procedures. Seedlings that germinated exhibited robust phototropic curvature. Frozen plant samples were returned on three space shuttle missions, and improvements in cold stowage and handing procedures in the second and third missions resulted in quality RNA extracted from the seedlings that was used in subsequent microarray analyses. While the TROPI experiment had technical and logistical difficulties, most of the procedures worked well due to refinement during the project.     

Thin film bioreactors in space.

Studies from the Skylab, SL-3 and D-1 missions have demonstrated that biological organisms grown in microgravity have changes in basic cellular functions such as DNA, mRNA and protein synthesis, cytoskeleton synthesis, glucose utilization and cellular differentiation. Since microgravity could affect prokaryotic and eukaryotic cells at a subcellular and molecular level, space offers us an opportunity to learn more about basic biological systems with one important variable removed. The thin film bioreactor will facilitate the handling of fluids in microgravity, under constant temperature and will allow multiple samples of cells to be grown with variable conditions. Studies on cell cultures grown in microgravity would enable us to identify and quantify changes in basic biological function in microgravity which are needed to develop new applications of orbital research and future biotechnology.     

Clinorotation reduces number, but not size, of cartilaginous nodules formed in micromass cultures of mouse limbbud cells.

In previous studies we used a ground based model to investigate the cellular responses to microgravity by exposing micromass cultures of embryonic limb cells to simulated weightlessness on a clinostat. Cultures set up in T-flasks and rotated at 30 rpm showed that clinostatted cultures had less chondrocyte differentiation than stationary or rotation controls, as assessed by number of nodules/culture stained with cartilage specific Alcian blue. In the current study, nodule size and shape of these nodules was assessed by interactive measurement of area, perimeter, circularity, and equivalent diameters, using the Optimas imaging software. Results show no significant difference in any of the measurements, indicating that clinorotation has no effect on expansion of the nodules either by differentiation of cells within the nodule, or by recruitment of cells into the nodule. The reduction in number of nodules without an alteration in size and shape indicates that the effect of simulated microgravity is to reduce the cell interactions required for the initial condensation of cells into a nodule, probably by interference with cell adhesion molecules.     

Chromosome aberrations in astronauts

A review of currently available data on in vivo induced chromosome damage in the blood lymphocytes of astronauts proves that cytogenetic biodosimetry analyses on blood collected within a week or two of return from space provides a reliable estimate of equivalent radiation dose and risk after protracted exposure to space radiation of a few months or more. Recent studies indicate that biodosimetry estimates from single spaceflights lie within the range expected from physical dosimetry and biophysical models, but very large uncertainties are associated with single individual measurements and the total sample population remains low. Retrospective doses may be more difficult to estimate because of the fairly rapid time-dependent loss of “stable” aberrations in blood lymphocytes. Also, biodosimetry estimates from individuals who participate in repeated missions, or very long (interplanetary) missions, may be complicated by an adaptive response to space radiation and/or changes in lymphocyte survival and repopulation. A discussion of published data is presented and specific issues related to space radiation biodosimetry protocols are discussed.     

Medical survey of European astronauts during Mir missions

This paper reviews the medical operations performed on six European astronauts during seven space missions on board the space station Mir. These missions took place between November 1988 and August 1999, and their duration ranged from 14 days to 189 days. Steps of pre-flight medical selection and flight certification are presented. Countermeasures program used during the flight, as well as rehabilitation program following short and long-duration missions are described. Also reviewed are medical problems encountered during the flight, post-flight physiological changes such as orthostatic intolerance, exercise capacity, blood composition, muscle atrophy, bone density, and radiation exposure.     

Embryogenesis, hatching and larval development of Artemia during orbital spaceflight.

Developmental biology studies, using gastrula-arrested cysts of the brine shrimp Artemia franciscana, were conducted during two flights of the space shuttle Atlantis (missions STS-37 and STS-43) in 1991. Dehydrated cysts were activated, on orbit, by addition of salt water to the cysts, and then development was terminated by the addition of fixative. Development took place in 5 ml syringes, connected by tubing to activation syringes, containing salt water, and termination syringes, containing fixative. Comparison of space results with simultaneous ground control experiments showed that equivalent percentages of naupliar larvae hatched in the syringes (40%). Thus, reactivation of development, completion of embryogenesis, emergence and hatching took place, during spaceflight, without recognizable alteration in numbers of larvae produced. Post-hatching larval development was studied in experiments where development was terminated, by introduction of fixative, 2 days, 4 days, and 8 days after reinitiation of development. During spaceflight, successive larval instars or stages, interrupted by molts, occurred, generating brine shrimp at appropriate larval instars. Naupliar larvae possessed the single naupliar eye, and development of the lateral pair of adult eyes also took place in space. Transmission electron microscopy revealed extensive differentiation, including skeletal muscle and gut endoderm, as well as the eye tissues. These studies demonstrate the potential value of Artemia for developmental biology studies during spaceflight, and show that extensive degrees of development can take place in this microgravity environment.     

3UPS-S并联机构单支链驱动奇异分析

并联机构进行单支链驱动时,由于受到其他支链的约束,不能实现全行程范围运动.为了分析并联机构单支链驱动受限的本质原因,采用旋量互易积的方法,计算单支链驱动的轴向速度分量.从计算结果可知,轴向速度为零时,单支链驱动到达边界.并联机构在输入空间发生驱动奇异,同时在工作空间发生位姿奇异,因此并联机构的驱动奇异就是位姿奇异.通过分析输入输出速度关系,并联机构到达输入空间边界.最后给出了并联机构单支链驱动的一般策略.      

航天器用可变发射率热控器件的研究进展

随着航天技术的发展,卫星的微型化对热控技术提出了挑战。可变发射率热控器件作为一种重要的航天器热控技术,对于航天器减小负载和体积,适应复杂多变的空间热环境具有重要的意义。基于热致变色技术的智能可变发射率热控器件可以根据环境温度实现智能热控,其结构简单,能最大限度地减小热控系统的体积和质量,是一种非常有潜力的航天器热控技术。概述了主动型和被动型两类可变发射率热控器件的基本原理和进展,并对钒氧化物基热致变色可变发射率热控器件的研究进展、存在问题予以了重点介绍,展望了未来航天器用可变发射率热控器件的发展趋势。