“Newton’s cradle” proton relay with amide–imidic acid tautomerization in inverting cellulase visualized by neutron crystallography

by on 22 August 2016in Biology & Biotechnology No comment

Hydrolysis of carbohydrates is a major bioreaction in nature, catalyzed by glycoside hydrolases (GHs). We used neutron diffraction and high-resolution x-ray diffraction analyses to investigate the hydrogen bond network in inverting cellulase PcCel45A, which is an endoglucanase belonging to subfamily C of GH family 45, isolated from the basidiomycete Phanerochaete chrysosporium. Examination of the enzyme and enzyme-ligand structures indicates a key role of multiple tautomerizations of asparagine residues and peptide bonds, which are finally connected to the other catalytic residue via typical side-chain hydrogen bonds, in forming the “Newton’s cradle”–like proton relay pathway of the catalytic cycle. Amide–imidic acid tautomerization of asparagine has not been taken into account in recent molecular dynamics simulations of not only cellulases but also general enzyme catalysis, and it may be necessary to reconsider our interpretation of many enzymatic reactions.

Related URLs:
http://advances.sciencemag.org/content/1/7/e1500263

The actin cytoskeleton is a suppressor of the endogenous skewing behaviour of Arabidopsis primary roots in microgravity

by on 22 August 2016in Biology & Biotechnology No comment

Before plants can be effectively utilised as a component of enclosed life-support systems for space exploration, it is important to understand the molecular mechanisms by which they develop in microgravity. Using the Biological Research in Canisters (BRIC) hardware on board the second to the last flight of the Space Shuttle Discovery (STS-131 mission), we studied how microgravity impacts root growth in Arabidopsis thaliana. Ground-based studies showed that the actin cytoskeleton negatively regulates root gravity responses on Earth, leading us to hypothesise that actin might also be an important modulator of root growth behaviour in space. We investigated how microgravity impacted root growth of wild type (ecotype Columbia) and a mutant (act2-3) disrupted in a root-expressed vegetative actin isoform (ACTIN2). Roots of etiolated wild-type and act2-3 seedlings grown in space skewed vigorously toward the left, which was unexpected given the reduced directional cue provided by gravity. The left-handed directional root growth in space was more pronounced in act2-3 mutants than wild type. To quantify differences in root orientation of these two genotypes in space, we developed an algorithm where single root images were converted into binary images using computational edge detection methods. Binary images were processed with Fast Fourier Transformation (FFT), and histogram and entropy were used to determine spectral distribution, such that high entropy values corresponded to roots that deviated more strongly from linear orientation whereas low entropy values represented straight roots. We found that act2-3 roots had a statistically stronger skewing/coiling response than wild-type roots, but such differences were not apparent on Earth. Ultrastructural studies revealed that newly developed cell walls of space-grown act2-3 roots were more severely disrupted compared to space-grown wild type, and ground control wild-type and act2-3 roots. Collectively, our results provide evidence that, like root gravity responses on Earth, endogenous directional growth patterns of roots in microgravity are suppressed by the actin cytoskeleton. Modulation of root growth in space by actin could be facilitated in part through its impact on cell wall architecture.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/23952736

Orthostatic Intolerance After ISS and Space Shuttle Missions

by on 22 August 2016in Biology & Biotechnology No comment

INTRODUCTION: Cardiovascular deconditioning apparently progresses with flight duration, resulting in a greater incidence of orthostatic intolerance following long-duration missions. Therefore, we anticipated that the proportion of astronauts who could not complete an orthostatic tilt test (OTT) would be higher on landing day and the number of days to recover greater after International Space Station (ISS) than after Space Shuttle missions. METHODS: There were 20 ISS and 65 Shuttle astronauts who participated in 10-min 80 degrees head-up tilt tests 10 d before launch, on landing day (R+0), and 3 d after landing (R+3). Fisher’s Exact Test was used to compare the ability of ISS and Shuttle astronauts to complete the OTT. Cox regression was used to identify cardiovascular parameters associated with OTT completion and mixed model analysis was used to compare the change and recovery rates between groups. RESULTS: The proportion of astronauts who completed the OTT on R+0 (2 of 6) was less in ISS than in Shuttle astronauts (52 of 65). On R+3, 13 of 15 and 19 of 19 of the ISS and Shuttle astronauts, respectively, completed the OTT. An index comprised of stroke volume and diastolic blood pressure provided a good prediction of OTT completion and was altered by spaceflight similarly for both astronaut groups, but recovery was slower in ISS than in Shuttle astronauts. CONCLUSIONS: The proportion of ISS astronauts who could not complete the OTT on R+0 was greater and the recovery rate slower after ISS compared to Shuttle missions. Thus, mission planners and crew surgeons should anticipate the need to tailor scheduled activities and level of medical support to accommodate protracted recovery after long-duration microgravity exposures.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/26630196

The scoring quality of astronauts’ sleeps using Fuzzy C-Means (FCM) during microgravity effect in the International Space Station (ISS)

by on 22 August 2016in Biology & Biotechnology No comment

For successful health programme of Japanese astronauts during space activity in the International Space Station (ISS), we looked into the sleep quality of astronauts by using Fuzzy C-Means (FCM). At the first step, we collected observation data by following sleep schedule of astronaut Noguchi for fifteen days (26 July 2005 to 9 August 2005) during STS-114 spaceflight. The Fuzzy C-Mean (FCM) was used to design the score quality of astronauts’ sleeps. We defined three condition of sleep quality (e.g. sleepy, sleep, deep sleep) with the minimum score (zero) and maximum score (nine). Based on the preliminary result, we found that the minimum sleep score of astronaut Noguchi was from 7 to 9 August 2005 which was due to landing process and the maximum sleep score we found was later than 27 July 2005 during docking on ISS.

Related URLs:
http://www.kemalapublisher.com/index.php/ppi-ukm/article/view/28

In vitro organogenesis from pluripotent stem cells

by on 22 August 2016in Biology & Biotechnology No comment

Pluripotent stem cells (PSCs) have the ability to spontaneously generate structured tissues in vitro reminiscent of embryonic tissue development. Recently, complex organoids such as cortical tissues, cerebral brain organoids, optical cups, intestinal tissues, and liver buds have been generated from PSCs derived from healthy individuals and patients with genetic diseases, providing powerful tools to understand morphogenesis and disease pathology. This article highlights recent advances in the state-of-art generation of organoids from PSCs, possible signaling pathways and mechanisms involved in organogenesis, and the understanding of extracellular microenvironment. Challenges involved in the organoid generation such as increasing organoid size, enhancing the tissue complexity, and improving functional maturation are also discussed.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/24762764

Isokinetic Strength Changes Following Long-Duration Space ight on the ISS

by on 22 August 2016in Biology & Biotechnology No comment

Long-duration spaceflight results in a loss of muscle strength that poses both operational and medical risks, particularly during emergency egress, upon return to Earth, and during future extraterrestrial exploration. Isokinetic testing of the knee, ankle, and trunk quantifies movement-specific strength changes following spaceflight and offers insight into the effectiveness of in-flight exercise countermeasures. We retrospectively evaluated changes in isokinetic strength for 37 ISS crewmembers (Expeditions 1–25) following 163 +/- 38 d (mean +/- SD) of spaceflight. Gender, in-flight resistance exercise hardware, and preflight strength were examined as potential modifiers of spaceflight-induced strength changes. Mean isokinetic strength declined 8–17% following space ight. One month after return to Earth, strength had improved, but small defcits of 1–9% persisted. Space flight-induced strength losses were not different between men and women. Mean strength losses were as much as 7% less in crewmembers who flew after the Advanced Resistive Exercise Device (ARED) replaced the interim Resistive Exercise Device (iRED) as the primary in-flight resistance exercise hardware, although these differences were not statistically significant. Absolute and relative preflight strength were moderately correlated (r= -0.47 and -0.54, respectively) with post flight strength changes. In-flight resistance exercise did not prevent decreased isokinetic strength after long-duration spaceflight. However, continued utilization of ARED, a more robust resistance exercise device providing higher loads than iRED, may result in greater benefits as exercise prescriptions are optimized. With reconditioning upon return to Earth, strength is largely recovered within 30 d.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/26630197

Osteoprotegerin is an effective countermeasure for spaceflight-induced bone loss in mice

by on 22 August 2016in Biology & Biotechnology No comment

Bone loss associated with microgravity exposure poses a significant barrier to long-duration spaceflight. Osteoprotegerin-Fc (OPG-Fc) is a receptor activator of nuclear factor kappa-B ligand (RANKL) inhibitor that causes sustained inhibition of bone resorption after a single subcutaneous injection. We tested the ability of OPG-Fc to preserve bone mass during 12 days of spaceflight (SF). 64-day-old female C57BL/6J mice (n=12/group) were injected subcutaneously with OPG-Fc (20mg/kg) or an inert vehicle (VEH), 24h prior to launch. Ground control (GC) mice (VEH or OPG-Fc) were maintained under environmental conditions that mimicked those in the space shuttle middeck. Age-matched baseline (BL) controls were sacrificed at launch. GC/VEH, but not SF/VEH mice, gained tibia BMD and trabecular volume fraction (BV/TV) during the mission (P<0.05 vs. BL). SF/VEH mice had lower BV/TV vs. GC/VEH mice, while SF/OPG-Fc mice had greater BV/TV than SF/VEH or GC/VEH. SF reduced femur elastic and maximum strength in VEH mice, with OPG-Fc increasing elastic strength in SF mice. Serum TRAP5b was elevated in SF/VEH mice vs. GC/VEH mice. Conversely, SF/OPG-Fc mice had lower TRAP5b levels, suggesting that OPG-Fc preserved bone during spaceflight via inhibition of osteoclast-mediated bone resorption. Decreased bone formation also contributed to the observed osteopenia, based on the reduced femur periosteal bone formation rate and serum osteocalcin level. Overall, these observations suggest that the beneficial effects of OPG-Fc during SF are primarily due to dramatic and sustained suppression of bone resorption. In growing mice, this effect appears to compensate for the SF-related inhibition of bone formation, while preventing any SF-related increase in bone resorption. We have demonstrated that the young mouse is an appropriate new model for SF-induced osteopenia, and that a single pre-flight treatment with OPG-Fc can effectively prevent the deleterious effects of SF on mouse bone. Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/26318907

Physical Training for Long-Duration Spaceflight

by on 22 August 2016in Biology & Biotechnology No comment

INTRODUCTION: Physical training has been conducted on the International Space Station (ISS) for the past 10 yr as a countermeasure to physiological deconditioning during spaceflight. Each member space agency has developed its own approach to creating and implementing physical training protocols for their astronauts. We have divided physical training into three distinct phases (preflight, in-flight, and postflight) and provided a description of each phase with its constraints and limitations. We also discuss how each member agency (NASA, ESA, CSA, and JAXA) prescribed physical training for their crewmembers during the first 10 yr of ISS operations. It is important to understand the operational environment, the agency responsible for the physical training program, and the constraints and limitations associated with spaceflight to accurately design and implement exercise training or interpret the exercise data collected on ISS. As exploration missions move forward, resolving agency differences in physical training programs will become important to maximizing the effectiveness of exercise as a countermeasure and minimizing any mission impacts.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/26630191

Understanding the effects of spaceflight on head–trunk coordination during walking and obstacle avoidance

by on 22 August 2016in Biology & Biotechnology No comment

Prolonged exposure to spaceflight conditions results in a battery of physiological changes, some of which contribute to sensorimotor and neurovestibular deficits. Upon return to Earth, functional performance changes are tested using the Functional Task Test (FTT), which includes an obstacle course to observe post-flight balance and postural stability, specifically during turning. The goal of this study was to quantify changes in movement strategies during turning events by observing the latency between head-and-trunk coordinated movements. It was hypothesized that subjects experiencing neurovestibular adaptations would exhibit head-to-trunk locking (‘en bloc’ movement) during turning, exhibited by a decrease in latency between head and trunk movement. FTT data samples were collected from 13 ISS astronauts and 26 male 70-day head down tilt bed rest subjects, including bed rest controls (10 BRC) and bed rest exercisers (16 BRE). Samples were analyzed three times pre-exposure, immediately post-exposure (0 or 1-day post) and 2–3 times during recovery from the unloading environment. Two 3D inertial measurements units (XSens MTx) were attached to subjects, one on the head and one on the upper back. This study focused primarily on the yaw movements about the subject’s center of rotation. Time differences (latency) between head and trunk movement were averaged across a slalom obstacle portion, consisting of three turns (approximately three 601 turns). All participants were grouped as ‘decreaser’ or ‘increaser’, relating to their change in head-to-trunk movement latency between pre- and post-environmental adaptation measures. Spaceflight unloading (ISS) showed a bimodal response between the ‘increaser’ and ‘decreaser’ group, while both bed rest control (BRC) and bed rest exercise (BRE) populations showed increased preference towards a ‘decreaser’ categorization, displaying greater head–trunk locking. It is clear that changes in movement strategies are adopted during exposure to an unloading environment. These results further the under- standing of vestibular–somatosensory convergence and support the use of bed rest as an exclusionary model to better understand sensorimotor changes in spaceflight.

Related URLs:
http://ntrs.nasa.gov/search.jsp?R=20140012973

Optic disc edema in an astronaut after repeat long-duration space flight

by on 22 August 2016in Biology & Biotechnology No comment

BACKGROUND: A number of ophthalmic findings including optic disc edema, globe flattening, and choroidal folds have been observed in several astronauts after long-duration space flights. The authors report the first astronaut with previously documented postflight ophthalmic abnormalities who developed new pathological changes after a repeat long-duration mission. METHODS: A case study of an astronaut with 2 long-duration (6 months) exposures to microgravity. Before and after his first long-duration space flight, he underwent complete eye examination, including fundus photography. Before and after his second flight, 9 years later, he underwent fundus photography, optical coherence tomography, ocular ultrasonography, and brain magnetic resonance imaging, as well as in-flight fundus photography and ultrasound. RESULTS: After his first long-duration mission, the astronaut was documented to have eye findings limited to unilateral choroidal folds and a single cotton wool spot. During a subsequent 6-month mission, he developed more widespread choroidal folds and new onset of optic disc edema in the same eye. CONCLUSION: Microgravity-induced anatomical changes that occurred during the first mission may have set the stage for recurrent or additional changes when the astronaut was subjected to physiological stress of repeat space flight.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/23851997