The biological clock of Neurospora in a microgravity environment.

The circadian rhythm of conidiation in Neurospora crassa is thought to be an endogenously derived circadian oscillation; however, several investigators have suggested that circadian rhythms may, instead, be driven by some geophysical time cue(s). An experiment was conducted on space shuttle flight STS-9 in order to test this hypothesis; during the first 7-8 cycles in space, there were several minor alterations observed in the conidiation rhythm, including an increase in the period of the oscillation, an increase in the variability of the growth rate and a diminished rhythm amplitude, which eventually damped out in 25% of the flight tubes. On day seven of flight, the tubes were exposed to light while their growth fronts were marked. Some aspect of the marking process reinstated a robust rhythm in all the tubes which continued throughout the remainder of the flight. These results from the last 86 hours of flight demonstrated that the rhythm can persist in space. Since the aberrant rhythmicity occurred prior to the marking procedure, but not after, it was hypothesized that the damping on STS-9 may have resulted from the hypergravity pulse of launch. To test this hypothesis, we conducted investigations into the effects of altered gravitational forces on conidiation. Exposure to hypergravity (via centrifugation), simulated microgravity (via the use of a clinostat) and altered orientations (via alterations in the vector of a 1 g force) were used to examine the effects of gravity upon the circadian rhythm of conidiation.     

The expression of a circadian rhythm in two strains of Chlamydomonas reinhardii in space.

During the D1 mission the endogenous circadian rhythm of the photoaccumulation response persisted in two strains of Chlamydomonas. The amplitude was about twice as high in space as on the ground indicating that a larger fraction of cells was able to contribute to the expression of the rhythm. On the ground, cells usually enter the light cone of the illuminated area in the recording cuvette on the upper edge and leave it, due to gravity, on the lower one in a pulsating manner. This sometimes produces high frequency oscillations of light extinction on the ground. In space there were no such fluctuations; instead, cells swam into the light and stayed there harvesting more light energy for photosynthesis than did control cells. This probably enhanced the survival rate and increased the fraction of motile cells which contribute to the photoaccumulation. A more sophisticated evaluation technique allowed determination of the phase in the short period strain; it was delayed by two hours compared to the control. In an acetate free wildtype sample a rhythm with a period of about 24 hours was also detected.     

24小时头低位卧床对人体生理功能的影响

为了探讨航天飞行初期人体生理功能的变化及“空间适应性综合症”的机理,实验观察了12名受试者在头低位6°卧床期间心率、呼吸、体温和脑电的变化。实验中所有指标均同时记录在卧床实验室配套的生理测试处理系统上,并用此系统对各生理指标的变化进行计算机分析。分析结果表明头低位卧床中人体内的一些重要生理功能如心率、呼吸、脑电的昼夜节律仍存在,但出现一定的紊乱,表现在:①心率昼夜变化双峰波消失或模糊,时相推迟。②体温昼夜节律变化不明显;额部皮肤温度无明显的昼夜节律变化,腿部皮肤温度在卧床初期明显增加,之后波动较大。③卧床中脑电α波段功率谱值下降,而δ、θ波功率谱值上升。卧床中R-R 间期频谱分析和脑电功率谱分析的结果表明卧床中心脑的调节和皮层的兴奋性也发生一定变化。     

Circadian rhythms in a long-term duration space flight.

In order to maintain cosmonaut health and performance, it is important for the work-rest schedule to follow human circadian rhythms (CR). What happens with CR in space flight? Investigations of CR in mammals revealed, that the circadian phase in flight is less stable, probably due to a displacement of the range of entrainment, resulting from internal period change (the latter was confirmed on insects). The circadian period may be a gravity-dependent parameter. If so, the basic biological requirement for the day length might be different in weightlessness. On this basis, a higher risk of desynchronosis is expected in a long-duration space flight. As a countermeasure, a non-24-hr day length could be suggested, being close to the internal circadian period (in humans about 25 hr). Taking into account a possible displacement of period in weightlessness, it seems reasonable to establish a flexible work-rest schedule, capable to follow the body temperature CR by means of biofeedback.     

空间里的时间:微重力等环境下的生物节律研究

生物钟是地球上的生物为适应环境周期性变化经历长期演化而来的内在机制.在分子水平上受生物钟基因及其他相关基因的调节;在组织水平上,生物钟由主生物钟和外周生物钟组成.生物钟对于各种生物的生理、认知和行为等具有重要功能,是生物适应环境的决定因素之一.空间环境下的微重力、辐射、光照条件、社会性因素等与地面存在很大差异,这些因素均可能导致节律紊乱,影响生物的生理及环境适应性.因此,对地外生命的研究也应该考虑生物钟因素.对航天员而言,节律紊乱可引起睡眠障碍,并且对骨肌系统、神经系统、心血管系统及内分泌系统等造成不利影响,导致人的认知和工效水平下降.在未来空间生命探索以及航天员健康保障研究中,生物钟是一个不可忽视的重要因素.      

Gravitational biology and the mammalian circadian timing system.

Mammals have evolved under the influence of many selective pressures. Two of these pressures have been the static force of gravity and the daily variations in the environment due to the rotation of the earth. It is now clear that each of these pressures has led to specific adaptations which influence how organisms respond to changes in either gravity or daily time cues. However, several unpredicted responses to altered gravitational environments occur within the homeostatic and circadian control systems. These results may be particularly relevant to biological and medical issues related to spaceflight. This paper demonstrates that the homeostatic regulation of rat body temperature, heart rate, and activity become depressed following exposure to a 2 G hyperdynamic field, and recovers within 5-6 days. In addition, the circadian rhythms of these same variables exhibit a depression of rhythm amplitude; however, recovery required a minimum of 7 days.     

Influence of gravity on the circadian timing system.

The circadian timing system (CTS) is responsible for daily temporal coordination of physiological and behavioral functions both internally and with the external environment. Experiments in altered gravitational environments have revealed changes in circadian rhythms of species ranging from fungi to primates. The altered gravitational environments examined included both the microgravity environment of spaceflight and hyperdynamic environments produced by centrifugation. Acute exposure to altered gravitational environments changed homeostatic parameters such as body temperature. These changes were time of day dependent. Exposure to gravitational alterations of relatively short duration produced changes in both the homeostatic level and the amplitude of circadian rhythms. Chronic exposure to a non-earth level of gravity resulted in changes in the period of the expressed rhythms as well as in the phase relationships between the rhythms and between the rhythms and the external environment. In addition, alterations in gravity appeared to act as a time cue for the CTS. Altered gravity also affected the sensitivity of the pacemaker to other aspects of the environment (i.e., light) and to shifts of time cues. Taken together, these studies lead to the conclusion that the CTS is indeed sensitive to gravity and its alterations. This finding has implications for both basic biology and space medicine.     

Transfer of space science and technology: A third world point of view

In contrast to the more common discussions on the how, where and when of transfer of space science and technology, we open a general discussion on the consequences of the space era in the developing countries and draw attention to the fact that space science and technology create a scientific and economic dependence on the industrially developed countries. Recommendations are made to render the space programs more beneficial to the Third World. These include : formation of local groups of high level space scientists and experts; instead of sophisticated transferred technology the use of space technologies appropriate to local economic and social status; services to the poorest fraction of the population and educational programs that protect the indigenous cultures.     

Temporal variation of the arterial pressure in healthy young people and its relation to geomagnetic activity in Mexico

We present a study of the temporal behavior of the systolic (SBP) and diastolic (DBP) blood pressure for a sample of 51 normotensive, healthy volunteers, 18 men and 33 women with an average age of 19 years old in Mexico City, Mexico, during April and May, 2008. We divided the data by sex along the circadian rhythm. Three geomagnetic storms occurred during the studied time-span. The strongest one, a moderate storm, is attributed to a coronal hole border that reached the Earth. The ANOVA test applied to the strongest storm showed that even though we are dealing with a moderate geomagnetic storm, there are statistically significant responses of the blood pressure. The superposed epoch analysis during a three-day window around the strongest storm shows that on average the largest changes occurred for the SBP. Moreover, the SBP largest increases occurred two days before and one day after this storm, and women are the most sensitive group as they present larger SBP and DBP average changes than men. Finally, given the small size of the sample, we cannot generalize our results.     

Planetarium Inversum -- a space vision for Earth education.

In a planetarium, the visitor is sitting on Earth and looking into an imaginary space. The Planetarium Inversum is the opposite: visitors are sitting in a space station, looking down on Mother Earth. It is a scientifically-based information show with visitors involvement, its elements being partially virtual (Earth in space has to be projected with highest possible resolution) but also containing real structures, such as the visitors' Earth observatory with adjacent biological systems (plant cultures and other ecological life support components). Its main message concerns the limits and the vulnerability of our home planet, its uniqueness, beauty and above all, its irreplaceableness: Earth does not have an emergency exit. The Earth observatory is part of a ring shaped, rotating space station of the type designed by Wernher von Braun decades ago. Visitors are told that gravity is being substituted by centrifugal force. Both types of life support systems are being demonstrated--self regenerative life based ones and technical ones as a backup (solar electric splitting of water and chemical absorption of respiratory CO2).     

Dose rate and repair effects on cell damage in Earth orbit.

Radiobiology experiments performed in space will encounter continuous exposures to the cosmic rays and fractionated exposures to trapped protons which accumulate to several hundred dose fractions in a few weeks. Using models of track structure and cellular kinetics combined with models of the radiation environment and radiation transport, we consider calculations of damage rates for cell cultures. Analysis of the role of repair mechanisms for space exposures for the endpoints of survival and transformation is emphasized.     

Gravimorphogenesis and ultrastructure of the fungus Flammulina velutipes grown in space, on clinostats and under hyper-g conditions.

The D-2-mission provided the facilities to cultivate the higher basidomycete Flammulina velutipes (Agaricales) in space for about 8 days. Gravimorphogenesis of developing fruiting body primordia in weightlessness was documented in comparison to cultures incubated on a 1xg reference centrifuge in space. Chemical fixation of fruiting bodies took place for later ultrastructural analysis. The microgravity grown fruiting bodies exhibited random orientation compared to the 1xg-cultures where fruiting bodies showed exactly negative gravitropic orientation. Weightlessness did not impair fruiting body morphogenesis and growth although flat and helically twisted stipes were observed. Ultrastructural analyses of microgravity-, 1xg- and 20xg-samples did not reveal sedimentable cell components. Gravitropic bending involves growth inhibition at the upper side of a horizontally oriented transition zone, the graviperceptive region of the stipe. The fastest ultrastructural response to the altered direction of the accelerational force is the accumulation of cytosolic vesicles at the lower part of this region. They contribute to the expansion of the central vacuole and therefore to the differential enlargement of the lower side of the stipe.     

Mortality and morphological anomalies related to the passage of cosmic heavy ions through the smallest flowering aquatic plant Wolffia arrhiza

Radiobiological effects of single cosmic heavy ions on individual, actively metabolizing test organisms, plants of Wolffia arrhiza, have been explored in an experiment flown aboard the Russian Biosatellite 10. Mortality induced during space flight, population dynamics during subsequent cultivation, and morphological anomalies occurring in the plants of these cultures were investigated. Correlation of these effects with the passage of a heavy ion was achieved by inserting monolayers of plants in a stack of surrounding plastic nuclear track detectors (BIO-STACK). Enhanced initial mortality and delayed decline of induced anomalies have been significantly associated with the passage of single heavy ions, in particular if ions penetrated the budding region of the plants. The prolonged persistence of anomalies in filial generations as an indication of delayed genetic damage has been detected for the first time as the consequence of the hit by a single heavy ion. Regarding radiation protection of space crew during prolonged missions, especially outside the magnetosphere, this appears to be a significant finding.     

In vivo and in vitro studies of cartilage differentiation in altered gravities.

The in vivo model our laboratory uses for studies of cartilage differentiation in space is the rat growth plate. Differences between missions, and in rat age and recovery times, provided differing results from each mission. However, in all missions, proliferation and differentiation of chondrocytes in the epiphyseal plate of spaceflown rats was altered as was matrix organization. In vitro systems, necessary complements to in vivo work, provide some advantages over the in vivo situation. In vitro, centrifugation of embryonic limb buds suppressed morphogenesis due to precocious differentiation, and changes in the developmental pattern suggest the involvement of Hox genes. In space, embryonic mouse limb mesenchyme cells differentiating in vitro on IML-1 had smoother membranes and lacked matrix seen in controls. Unusual formations, possibly highly ruffled membranes, were found in flight cultures. These results, coupled with in vivo centrifugation studies, show that in vivo or in vitro, the response of chondrocytes to gravitational changes follows Hert's curve as modified by Simon, i.e. decreased loading decreases differentiation, and increased loading speeds it up, but only to a point. After that, additional increases again slow down chondrogenesis.     

Somatic embryos of daylily in space

Poor growth and nuclear abnormalities observable in some space-grown plants have been hypothesized as due to a combination of factors such as degree of development, the specific way the plants are grown and the way they experience multiple stresses, some of which are space-specific. Data from a 132-day experiment on ‘Mir’ using embryogenic cell cultures of daylily (Hemerocallis) allow seemingly contradictory evidence from earlier Shuttle missions to be harmonized: a) the more developed an embryo the less likely it is to suffer catastrophic cell stress during growth, whereas the less developed it is, the greater its vulnerability; (b) the extent to which the stress becomes manifest is also dependent on the extent of pre-existing stresses imposed by suboptimal growing conditions; (c) an appropriate, albeit undesirable, ‘stress match’ with other non-equilibrium determinants, much like a ‘tug of war’, can result in genomic variations in space. It is not understood what is/are the feature(s) of the space environment that cause the various cell division perturbations but they have not yet been mimicked on earth. The stress symptoms were found only in space materials and, as predicted, they were most frequently encountered in smaller, less-developed materials grown under non-optimized conditions. It is concluded that, while any substantial deviation from ‘optimum’ can be a ‘stress’, spaceflight subjects vulnerable materials to cell division or DNA-repair stress(es) that appear distinctive, but remain elusive so far. Fastidiously-controlled growing environments must be devised to resolve the matter of direct versus indirect effects of space. On a practical level, it is predicted that adapting plant biotechnologies to space conditions will not be a casual matter.     

Radiation effects in nematodes: results from IML-1 experiments.

The nematode Caenorhabditis elegans was exposed to natural space radiation using the ESA Biorack facility aboard Spacelab on International Microgravity Laboratory 1, STS-42. For the major experimental objective dormant animals were suspended in buffer or on agar or immobilized next to CR-39 plastic nuclear track detectors to correlate fluence of HZE particles with genetic events. This configuration was used to isolate mutations in a set of 350 essential genes as well as in the unc-22 structural gene. From flight samples 13 mutants in the unc-22 gene were isolated along with 53 lethal mutations from autosomal regions balanced by a translocation eT1(III;V). Preliminary analysis suggests that mutants from worms correlated with specific cosmic ray tracks may have a higher proportion of rearrangements than those isolated from tube cultures on a randomly sampled basis. Right sample mutation rate was approximately 8-fold higher than ground controls which exhibited laboratory spontaneous frequencies.     

Biorhythms on cellular and organismic level (introduction).

The regular change of day and night, of light and darkness during millions of years has strongly affected the development of life on earth. Many organisms adapted themselves to this environmental condition and, finally, evolved an endogenous timer which usually is in phase with the earth's rotation and causes many functions to perform one oscillation per day. Such circadian rhythms (derived from circa dies i.e. about 1 day) were found in almost all classes of plants and animals, and even in protozoans. They persist in a constant environment and, therefore, are independent of any known external trigger signals. Since even unicells perform circadian rhythms which are similar to those observed in highly developed multicellular organisms many scientists favor the existence of a basic mechanism common to all kinds of biological clocks that is located somewhere in the single cell and probably comprises many different biochemical reactions. One purpose of this topical meeting was to discuss how organisms respond to the absence of gravity and terrestrial zeitgeber and how they may react to the imposing of hypergravity fields. Another aim was to develop model-mechanisms appropriate to describe these responses.     

Radiation-induced chromosomal instability in human mammary epithelial cells.

Karyotypes of human cells surviving X- and alpha-irradiation have been studied. Human mammary epithelial cells of the immortal, non-tumorigenic cell line H184B5 F5-1 M/10 were irradiated and surviving clones isolated and expanded in culture. Cytogenetic analysis was performed using dedicated software with an image analyzer. We have found that both high- and low-LET radiation induced chromosomal instability in long-term cultures, but with different characteristics. Complex chromosomal rearrangements were observed after X-rays, while chromosome loss predominated after alpha-particles. Deletions were observed in both cases. In clones derived from cells exposed to alpha-particles, some cells showed extensive chromosome breaking and double minutes. Genomic instability was correlated to delayed reproductive death and neoplastic transformation. These results indicate that chromosomal instability is a radiation-quality-dependent effect which could determine late genetic effects, and should therefore be carefully considered in the evaluation of risk for space missions.     

基于EEG信号特征的脑力疲劳快速检测方法

空间站飞行过程中航天员容易产生脑力疲劳，其是影响作业效率和引起失误的主要因素。为此，研究人体脑力疲劳的快速检测方法，将有利于保障在轨运行安全。脑电波(EEG)的特征变化能够反映出大脑疲劳状态，但现有EEG方法分析脑力疲劳时需要多个导联的信号，这严重限制了其在空间站环境中的实际应用。通过地基实验，采用36 h睡眠剥夺的方式成功诱发出45名受试者的多种脑力疲劳状态。针对EEG信号的非平稳性，设计的8层db4小波变换结构，有效分解出了δ、θ、α和β脑节律波。先使用方差分析( ANOVA)和Logistic回归筛选出脑力疲劳敏感特征，再依据脑力疲劳敏感特征数量进一步筛选出脑力疲劳敏感导联，应用6个敏感导联的特征分别构建了随机森林回归模型。加权融合6个导联处的回归模型，形成脑力疲劳快速检测模型，其平均精确率高达85.25%。      

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.