Adaptation and Changes in Actin Dynamics and Cell Motility as Early Responses of Cultured Mammalian Cells to Altered Gravitational Vector.

Cultured mammalian cells have been shown to respond to microgravity (µG), but the molecular mechanism is still unknown. The study we report here is focused on molecular and cellular events that occur within a short period of time, which may be related to gravity sensing by cells. Our assumption is that the gravity-sensing mechanism is activated as soon as cells are exposed to any new gravitational environment. To study the molecular events, we exposed cells to simulated µG (SµG) for 15 min, 30 min, 1 h, 2 h, 4 h, and 8 h using a three-dimensional clinostat and made cell lysates, which were then analyzed by reverse phase protein arrays (RPPAs) using a panel of 453 different antibodies. By comparing the RPPA data from cells cultured at 1G with those of cells under SµG, we identified a total of 35 proteomic changes in the SµG samples and found that 20 of these changes took place, mostly transiently, within 30 min. In the 4 h and 8 h samples, there were only two RPPA changes, suggesting that the physiology of these cells is practically indistinguishable from that of cells cultured at 1 G. Among the proteins involved in the early proteomic changes were those that regulate cell motility and cytoskeletal organization. To see whether changes in gravitational environment indeed activate cell motility, we flipped the culture dish upside down (directional change in gravity vector) and studied cell migration and actin cytoskeletal organization. We found that compared with cells grown right-side up, upside-down cells transiently lost stress fibers and rapidly developed lamellipodia, which was supported by increased activity of Ras-related C3 botulinum toxin substrate 1 (Rac1). The upside-down cells also increased their migratory activity. It is possible that these early molecular and cellular events play roles in gravity sensing by mammalian cells. Our study also indicated that these early responses are transient, suggesting that cells appear to adapt physiologically to a new gravitational environment.

Depression of Bone Density at the Weight-Bearing Joints in Wistar Hannover Rats by a Simulated Mechanical Stress Associated With Partial Gravity Environment.

The partial gravity environment in space can negatively affect bone health. This survey aimed to study the reaction of different parts of the lower limb bones of rats to partial gravity and the effects of different degrees of gravity on these bony parts. We used 15 8-week-old male Wistar Hannover rats were used at the beginning of the experiment. The degree of mechanical stress was modified, but the ankle joint was maintained at ∼30°, ∼120°, or ∼160° with or without plaster fixation during 10-day hindlimb suspension. Computed tomography was performed to measure the bone parameters [bone mineral density (BMD), trabecular BMD, cortical BMD, and cortical thickness] of each studied group of the whole, proximal, middle, and distal femur and distal tibia. BMD, trabecular BMD, and cortical thickness of the distal femur and proximal tibia of the simulated mechanical stress associated with partial gravity groups were significantly lower than those of the control group; the effect of different degrees of gravity on the same area of hindlimb bone had no significant difference. The simulated mechanical stress associated with partial gravity had the most significant effect on the bone close to the knee joint, with the largest weight-bearing response.

Combined Environment Simulator for Low-Dose-Rate Radiation and Partial Gravity of Moon and Mars.

Deep space exploration by humans has become more realistic, with planned returns to the Moon, travel to Mars, and beyond. Space radiation with a low dose rate would be a constant risk for space travelers. The combined effects of space radiation and partial gravity such as on the Moon and Mars are unknown. The difficulty for such research is that there are no good simulating systems on the ground to investigate these combined effects. To address this knowledge gap, we developed the Simulator of the environments on the Moon and Mars with Neutron irradiation and Gravity change (SwiNG) for in vitro experiments using disposable closed cell culture chambers. The device simulates partial gravity using a centrifuge in a three-dimensional clinostat. Six samples are exposed at once to neutrons at a low dose rate (1 mGy/day) using Californium-252 in the center of the centrifuge. The system is compact including two SwiNG devices in the incubator, one with and one without radiation source, with a cooling function. This simulator is highly convenient for ground-based biological experiments because of limited access to spaceflight experiments. SwiNG can contribute significantly to research on the combined effects of space radiation and partial gravity.

Simultaneous Exposure of Cultured Human Lymphoblastic Cells to Simulated Microgravity and Radiation Increases Chromosome Aberrations.

During space travel, humans are continuously exposed to two major environmental stresses, microgravity (µG) and space radiation. One of the fundamental questions is whether the two stressors are interactive. For over half a century, many studies were carried out in space, as well as using devices that simulated µG on the ground to investigate gravity effects on cells and organisms, and we have gained insights into how living organisms respond to µG. However, our knowledge on how to assess and manage human health risks in long-term mission to the Moon or Mars is drastically limited. For example, little information is available on how cells respond to simultaneous exposure to space radiation and µG. In this study, we analyzed the frequencies of chromosome aberrations (CA) in cultured human lymphoblastic TK6 cells exposed to X-ray or carbon ion under the simulated µG conditions. A higher frequency of both simple and complex types of CA were observed in cells exposed to radiation and µG simultaneously compared to CA frequency in cells exposed to radiation only. Our study shows that the dose response data on space radiation obtained at the 1G condition could lead to the underestimation of astronauts' potential risk for health deterioration, including cancer. This study also emphasizes the importance of obtaining data on the molecular and cellular responses to irradiation under µG conditions.

Development of a new image manipulation system based on detection of electroencephalogram signals from the operator's brain: a feasibility study.

Physicians require an adequate display system with a console within arm's reach to view images during surgical operations and interventional radiological examinations. However, manipulation of the console by physicians themselves may not be possible because their hands may be otherwise engaged. In this study, an image manipulation system using an electroencephalogram (EEG) sensor mounted on the operator's head was developed. In this system, data acquired by the device is used to manipulate images, and the output can be converted to commands for various actions such as paging, which can be controlled by the operator's eye-blink, and zooming of a region indicated by the cursor, which can be controlled by the operator's mental concentration. In this study, the MindWave Mobile headset was used as EEG sensor, and AZEWIN for the display system. Ten observers were enrolled and fitted with EEG device to determine the threshold values of blink strength and attention; threshold value of 100 for blink strength and 65 for attention were determined. Thirty-one observers were enrolled and fitted with EEG device to investigate average response-time; the average response time for detecting paging was 0.43 ± 0.02 s, and that for zooming was 5.85 ± 0.56 s. Thus, the proposed image manipulation system using the operator's EEG signals enabled physicians to assess and manipulate images without using their hands.