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At least 37 records · Page 2

Dual reporter transgene driven by 2.3Col1a1 promoter is active in differentiated osteoblasts

AIM: As quantitative and spatial analyses of promoter reporter constructs are not easily performed in intact bone, we designed a reporter gene specific to bone, which could be analyzed both visually and quantitatively by using chloramphenicol acetyltransferase (CAT) and a cyan version of green fluorescent protein (GFPcyan), driven by a 2.3-kb fragment of the rat collagen promoter (Col2.3). METHODS: The construct Col2.3CATiresGFPcyan was used for generating transgenic mice. Quantitative measurement of promoter activity was performed by CAT analysis of different tissues derived from transgenic animals; localization was performed by visualized GFP in frozen bone sections. To assess transgene expression during in vitro differentiation, marrow stromal cell and neonatal calvarial osteoblast cultures were analyzed for CAT and GFP activity. RESULTS: In mice, CAT activity was detected in the calvaria, long bone, teeth, and tendon, whereas histology showed that GFP expression was limited to osteoblasts and osteocytes. In cell culture, increased activity of CAT correlated with increased differentiation, and GFP activity was restricted to mineralized nodules. CONCLUSION: The concept of a dual reporter allows a simultaneous visual and quantitative analysis of transgene activity in bone.

Non-NASA Center

Hindlimb unloading alters ligament healing

We investigated the hypothesis that hindlimb unloading inhibits healing in fibrous connective tissue such as ligament. Male rats were assigned to 3- and 7-wk treatment groups with three subgroups each: sham control, ambulatory healing, and hindlimb-suspended healing. Ambulatory and suspended animals underwent surgical rupture of their medial collateral ligaments, whereas sham surgeries were performed on control animals. After 3 or 7 wk, mechanical and/or morphological properties were measured in ligament, muscle, and bone. During mechanical testing, most suspended ligaments failed in the scar region, indicating the greatest impairment was to ligament and not to bone-ligament insertion. Ligament testing revealed significant reductions in maximum force, ultimate stress, elastic modulus, and low-load properties in suspended animals. In addition, femoral mineral density, femoral strength, gastrocnemius mass, and tibialis anterior mass were significantly reduced. Microscopy revealed abnormal scar formation and cell distribution in suspended ligaments with extracellular matrix discontinuities and voids between misaligned, but well-formed, collagen fiber bundles. Hence, stress levels from ambulation appear unnecessary for formation of fiber bundles yet required for collagen to form structurally competent continuous fibers. Results support our hypothesis that hindlimb unloading impairs healing of fibrous connective tissue. In addition, this study provides compelling morphological evidence explaining the altered structure-function relationship in load-deprived healing connective tissue.

NASA Discipline Musculoskeletal

Bion-11 Spaceflight Mission

The Sensors 2000! Program, in support of the Space Life Sciences Payloads Office at NASA Ames Research Center developed a suite of bioinstrumentation hardware for use on the Joint US/Russian Bion I I Biosatellite Mission (December 24, 1996 - January 7, 1997). This spaceflight included 20 separate experiments that were organized into a complimentary and interrelated whole, and performed by teams of US, Russian, and French investigators. Over 40 separate parameters were recorded in-flight on both analog and digital recording media for later analysis. These parameters included; Electromyogram (7 ch), Electrogastrogram, Electrooculogram (2 ch), ECG/EKG, Electroencephlogram (2 ch), single fiber firing of Neurovestibular afferent nerves (7 ch), Tendon Force, Head Motion Velocity (pitch & yaw), P02 (in vivo & ambient), temperature (deep body, skin, & ambient), and multiple animal and spacecraft performance parameters for a total of 45 channels of recorded data. Building on the close cooperation of previous missions, US and Russian engineers jointly developed, integrated, and tested the physiologic instrumentation and data recording system. For the first time US developed hardware replaced elements of the Russian systems resulting in a US/Russian hybrid instrumentation and data system that functioned flawlessly during the 14 day mission.

Skidmore, M.

Life Sciences Space Station planning document: A reference payload for the Life Sciences Research Facility

The Space Station, projected for construction in the early 1990s, will be an orbiting, low-gravity, permanently manned facility providing unprecedented opportunities for scientific research. Facilities for Life Sciences research will include a pressurized research laboratory, attached payloads, and platforms which will allow investigators to perform experiments in the crucial areas of Space Medicine, Space Biology, Exobiology, Biospherics and Controlled Ecological Life Support System (CELSS). These studies are designed to determine the consequences of long-term exposure to space conditions, with particular emphasis on assuring the permanent presence of humans in space. The applied and basic research to be performed, using humans, animals, and plants, will increase our understanding of the effects of the space environment on basic life processes. Facilities being planned for remote observations from platforms and attached payloads of biologically important elements and compounds in space and on other planets (Exobiology) will permit exploration of the relationship between the evolution of life and the universe. Space-based, global scale observations of terrestrial biology (Biospherics) will provide data critical for understanding and ultimately managing changes in the Earth's ecosystem. The life sciences community is encouraged to participate in the research potential the Space Station facilities will make possible. This document provides the range and scope of typical life sciences experiments which could be performed within a pressurized laboratory module on Space Station.

Source record

scPlantAnnotate: an accurate and robust transformer-based model for plant cell type annotation

Accurate cell type annotation remains a major bottleneck in plant single-cell RNA sequencing (scRNA-seq), where existing tools are often adapted from animal studies and perform sub-optimally on plant data. The lack of plant-specific computational frameworks limits the construction of plant cell atlases and downstream biological discovery. We develop and evaluate scPlantAnnotate, a Transformer-based reference annotation framework tailored for plant scRNA-seq data, and benchmark it against state-of-the-art deep learning and conventional methods across multiple plant species. Species-specific scPlantAnnotate models were trained using curated datasets from Arabidopsis thaliana, Zea mays, Oryza sativa, and Glycine max. We compared scPlantAnnotate with leading baselines under both standard random-split evaluation and a more stringent leave-one-dataset-out setting, which tests robustness to completely unseen datasets and tissue types. scPlantAnnotate consistently outperforms existing approaches across all four species under random-split evaluation. In the leave-one-dataset-out setting for A. thaliana, where performance drops markedly for all methods due to strong batch effects and dataset heterogeneity, scPlantAnnotate nonetheless achieves the highest Accuracy, Macro-F1, Balanced Accuracy, and Macro-AUROC on average and ranks first on most held-out datasets. These results demonstrate improved robustness to dataset shifts, a critical yet underexplored challenge in plant scRNA-seq analysis. A freely accessible web server enables users to annotate their own datasets using pretrained models. scPlantAnnotate provides a plant-specific, Transformer-based framework for single-cell annotation that delivers state-of-the-art performance and enhanced robustness to unseen datasets. By addressing limitations of existing tools and enabling scalable reference-based annotation, scPlantAnnotate supports the development of comprehensive plant cell atlases and facilitates broader use of single-cell genomics in plant biology.

Bioinformatics

Waste to worth: a circular solution through lignin engineering

The inefficient management of industrial waste threatens environmental sustainability, requiring scalable resource recovery solutions. To address these challenges, we propose a novel and sustainable in-situ one-step strategy for the simultaneous recovery and functionalization of lignin from industrial black liquor, using a sustainable organic acid system assisted by choline chloride. The functionalized lignin adsorbent material (FLAM) exhibited superior adsorption performance, effectively treating animal feedlot wastewater to recover ammonia. The ammonia enriched FLAM was subsequently repurposed as a slow-release fertilizer, enhancing plant growth and demonstrating a closed-loop pathway for waste valorization and nutrient recycling. Methylene blue was used as a model cationic compound to simulate the adsorption behavior and interactions. FLAM achieved a high adsorption capacity of 725.98 mg/g across a broad pH range, significantly outperforming commercial kraft lignin. This sustainable approach advances circular economy principles by integrating remediation and resource recovery, offering a scalable, eco-friendly solution for environmental applications.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI

Producing cultivated meat cuts that match traditional meat in quality and affordability (CRADA Final Report)

Novel Farms, Inc., as a participant of the Cyclotron Road Lab-Embedded Entrepreneurship Program, investigated the performance of various animal cell lines and animal cell media formulations using its proprietary scaffolds as a growth substratum towards the development of economically-feasible cultivated meat production. As an alternative to conventionally farmed animal meat, cultivated meat production has the potential to yield a marginal carbon footprint while providing sustainable and cruelty-free animal protein. The prohibitively high cost of the growth factors necessary in animal cell culture media constitutes the primary barrier that hinders the economically viable production of cultivated meat products. Furthermore, scaffolding is required to create structured meat products, but current options are unsuitable for cost-effective large-scale production of cultivated meat. Novel Farms has developed a proprietary technology to produce scaffolding that has the potential to reduce production costs while providing an ideal 3D surface for cell growth. Developing cost-efficient methods for cultivated meat production will be vital in satisfying the growing demand for animal protein while avoiding the deforestation, carbon emissions, biodiversity loss and other negative outcomes of industrial animal farming.

59 BASIC BIOLOGICAL SCIENCES

The muscle spindle as a feedback element in muscle control

The muscle spindle, the feedback element in the myotatic (stretch) reflex, is a major contributor to muscular control. Therefore, an accurate description of behavior of the muscle spindle during active contraction of the muscle, as well as during passive stretch, is essential to the understanding of muscle control. Animal experiments were performed in order to obtain the data necessary to model the muscle spindle. Spectral density functions were used to identify a linear approximation of the two types of nerve endings from the spindle. A model reference adaptive control system was used on a hybrid computer to optimize the anatomically defined lumped parameter estimate of the spindle. The derived nonlinear model accurately predicts the behavior of the muscle spindle both during active discharge and during its silent period. This model is used to determine the mechanism employed to control muscle movement.

Andrews, L. T.

Efficiency characteristics of crescent-shaped wings and caudal fins

Caudal (tail) fins of fish and aquatic mammals that cruise long distances, and wings of certain birds, often have the shape of a crescent moon. This study investigates how the crescent shape contributes to the traveling performance of these animals. A steady-flow theory (Maskew, 1982) that correctly models the trailing wake was used to analyze lifting surface efficiency, which is dependent on the level of induced (or vortex) drag for a given lift and span of the lifting surface. This analysis shows that backward curvature of a wing improves induced efficiency to a value greater than that of the flat untwisted wing of elliptical shape considered optimal in classical wing theory (Prandt, 1921 and Munk, 1921). This increase of induced efficiency results from the nonplanar trailing vortex sheet produced by the crescent-shaped wing at a given angle of attack.

Van Dam, C. P.

A kinematic analysis of the Space Station remote manipulator system (SSRMS)

An efficient reverse analysis of three 6-degree-of-freedom (dof) subchains of the 7-dof SSRMS is presented. The first subchain is formed by locking the seventh joint. The second subchain is formed by locking the second joint, while the third subchain is formed by locking the first joint (the grounded joint is counted as the first joint in the chain). There are a maximum of eight different arm configurations in each of the three subchains, and these were determined by employing a computer-efficient algorithm, which required the rooting of only at most quadratic polynomials. The algorithms were implemented, and the SSRMS was employed in an animated environment to perform and practice a number of useful tasks for Space Station servicing. The locking of the second joint has the advantage in that an operator can choose the orientation of the plane that contains the two longest links so as to avoid collisions with obstacles. However, it has the disadvantage that when the second joint angle equals 0 deg or 180 deg, the manipulator is in a singularity configuration. This plane can also be oriented by specifying the first joint angle, so that the plane can be oriented arbitrarily and, in this, the singularity is avoided.

Crane, Carl D., III

Thermal degradation events as health hazards - Particle vs gas phase effects, mechanistic studies with particles

Experiments on animal subjects are performed to demonstrate that significant lung injury can result from the inhalation of ultrafine TiO2 or Al2O3 particles. The methods include intratracheal instillation of particles, long-term inhalation of particles, and in vitro studies of alveolar macrophages (AMs) to study the production of fibroplast growth factors. The ultrafine TiO2 particles are shown to induce more acute inflammatory reactions than larger particles and lead to persistent chronic effects in the AM-mediated clearance function of particles. The ultrafine particles also induce cytokines more readily, and the data generally suggests that the occurrence of such particles in thermal degradation events makes the fumes highly toxic. The exposure to thermal degradation products is therefore a critical concern for manned space missions with potentially degradable plastic products.

Oberdoerster, G.

Eccentric exercise training as a countermeasure to non-weight-bearing soleus muscle atrophy

This investigation tested whether eccentric resistance training could prevent soleus muscle atrophy during non-weight bearing. Adult female rats were randomly assigned to either weight bearing +/- intramuscular electrodes or non-weight bearing +/- intramuscular electrodes groups. Electrically stimulated maximal eccentric contractions were performed on anesthetized animals at 48-h intervals during the 10-day experiment. Non-weight bearing significantly reduced soleus muscle wet weight (28-31 percent) and noncollagenous protein content (30-31 percent) compared with controls. Eccentric exercise training during non-weight bearing attenuated but did not prevent the loss of soleus muscle wet weight and noncollagenous protein by 77 and 44 percent, respectively. The potential of eccentric exercise training as an effective and highly efficient counter-measure to non-weight-bearing atrophy is demonstrated in the 44 percent attenuation of soleus muscle noncollagenous protein loss by eccentric exercise during only 0.035 percent of the total non-weight-bearing time period.

Kirby, Christopher R.

Adaptation to Space: An Introduction

The cardiovascular and musculoskeletal systems are normally exposed to gradients of blood pressure and weight on Earth. These gradients increase blood pressure and tissue weight in dependent tissues of the body. Exposure to actual and simulated microgravity causes blood and tissue fluid to shift from the legs to the head. Studies of humans in space have documented facial edema, space motion sickness, decreased plasma volume, muscle atrophy, and loss of bone strength. Return of astronauts to Earth is accompanied by orthostatic intolerance, decreased neuromuscular coordination, and reduced exercise capacity. These factors decrease performance during descent from orbit and increase risk during emergency egress from the spacecraft. Models of simulated microgravity include 6 deg head-down tilt, immersion, and prolonged horizontal bedrest. Head-down tilt is the most accepted model and studies using this model of up to one year have been performed in Russia. Animal models which offer clear insights into the role of gravity on vertebrates include the developing giraffe and snakes from various habitats. Finally, possible countermeasures to speed readaptation of astronauts to gravity after prolonged space flight will be discussed.

Hargens, Alan R.

STS-112 Mission Highlights Resource Tape Part 2 of 3

An overview of the STS-112 Mission is presented. The fourth flight day begins with a view inside of the Destiny Laboratory of the International Space Station where Expedition Five Commander Valery Korzun is shown. The robotics workstation where Mission Specialist Sandra Mangus and Flight Engineer Peggy Whitson operate Canadarm 2 to lift the S(1) Truss segment out of the payload bay of the Space Shuttle Atlantis and maneuver it for installation onto the S(1) Truss is presented. Mission Specialist Piers Sellers is shown preparing for his six and one half hour spacewalk by performing breathing exercises. Animation of the installation of the S(1) Truss, and also the unbirthing of the S(1) truss is also presented. Mission Specialists David Wolf and Piers Sellers are shown getting suited for their spacewalks. During the spacewalk, David Wolf is removing the antenna assembly from its launched position and Piers Sellers is releasing launch restraints from the Radiator Beam Assembly. A beautiful view of the coast of Texas is captured during this spacewalk. Flight day five shows pilot Pam Melroy and Mission Specialist Dave Wolf working inside of the International Space Station's Quest Airlock. Flight day six is shown with Dave Wolf and Piers Sellers exiting their spacesuits inside of the Quest Airlock Module after a successful spacewalk. This presentation ends with views of the International Space Station's installed S(1) Truss, rotation of the radiator assembly and the radiator's coolant tubing.

Source record

Changes in gravity influence rat postnatal motor system development: from simulation to space flight

Our research examines the role of the environment in postnatal nervous system development. Recently we have been studying the effects of changes in gravity on the motor system of rats from postnatal day (P) 2 to 31 using kinematic analysis of swimming, walking, and righting reflexes. Using the tail suspension model of weightlessness we identified sensitive and critical periods of motor system development corresponding to the time during which a motor skill is first achieved. Motor performance in suspended animals was marked by slow swimming, walking, and air-righting, all of which were characterized by hindlimb extension. (Walton et al, Neurosci. 52,763,1992). The critical periods identified in these studies contributed to determining the age of animals for a small payload, NIH.R3. This 9-day mission (STS-72) included 2 litters at P5, P7, or P15 at launch. The P7-16 and P15-24 groups were studied post-flight. On the landing day (R+0) surface righting, swimming and walking were slower in flight compared to control animals. Differences were more marked in the younger animals and the hindlimbs were more affected than the forelimbs with marked, prolonged extension of, at least, the ankle joint angle. Readaptation to 1G was slower in the P7-16 group with righting reflexes adapting first, walking last. We have shown that gravity is an important factor in postnatal nervous system development and that its affect depends on the age of the animal, duration of the perturbation, and the motor function studied.

Non-NASA Center

Late ophthalmological complications after total body irradiation in non-human primates

PURPOSE: To investigate the long-term effects of total body irradiation (TBI) on the incidence and time course of ocular complications. MATERIALS AND METHODS: Rhesus monkeys treated with TBI photon doses up to 8.5 Gy and proton doses up to 7.5 Gy were studied at intervals up to 25 years post-irradiation. They were compared with control groups with a similar age distribution. Cataract formation and ocular fundus lesions were scored according to a standardized protocol. Fluorescein angiography and histopathology was performed in selected animals. RESULTS: Cataract formation occurred after a latent period of 3-5 years. Significant cataract induction was observed for photon-doses of 8 and 8.5 Gy and beyond 20 years after proton irradiation. The severity of the lesions represents significant impairment of vision and would require cataract surgery if similar results occurred in human bone marrow transplant patients. Fluorescein angiography demonstrated a normal pattern of retinal vessels in 13 out of 14 animals (93%) from the irradiated group and in eight out of nine animals (89%) from the control group. No additional lesions apart from age-related degenerative changes could be demonstrated. Histological evaluation revealed no radiation-associated vasculopathy. CONCLUSIONS: Radiation alone for doses up to 8.5 Gy of photons does not carry a potential risk for fundus pathology, whereas clinically important cataract induction should be anticipated within 5 years after photon doses of 8.0 and 8.5 Gy and proton doses in excess of 2.5 Gy.

NASA Discipline Radiation Health

Effect of 16-Day Spaceflight on the Morphology of Thick-Toed Geckos (Pachydactylus Bibronii Smith, 1846)

There are grounds to believe that space flown experiments on thick-toed geckos may help solve the problem of floatation of vertebrates in microgravity. Geckos of this species carry on the lower surface of their toes numerous setae, which allow them to remain attached to any surfaces regardless of the gravitational effects. Experiments were performed on 5 animals in each of the following groups: flight, basal, synchronous and laboratory controls. 32 hours after a 16- day flight the animals were euthanazed and examined using traditional histology and X-ray microtomography. Body mass losses were 10% in the flight animals, 7.4% in the synchronous controls, and 12.3% in the laboratory controls. Since the flight and synchronous animals were kept at 15-19 C, whereas the laboratory controls - at 26-28 C, it can be inferred that environmental temperatures impacted animal metabolism no less than flight induced stress. Blood tests of the flown animals showed a 12% decrease of erythrocytes and a 40% decrease of dark-nuclear granulocytes, with the number of light-nuclear granulocytes remaining unchanged. In the small intestine the number of goblet cells increased allowing them to occupy a large portion of the cyptal surface. Enhanced secretion was accompanied by the appearance of dead intestinal cells in the lumen. Clusters of degraded hepatocytes were found at the liver edges of flight animals. Signs of liver involution were similar to the changes produced by alcohol consumption but did not spread to its central part. In the heart, insignificant hypertrophy and excessive blood supply that still remained within the physiological norm were detected. No significant changes were found in the pancreas, lungs, nervous systems or the snouts of the flown animals, but the volume of their gallbladders was greater than in controls. The epithelium of toe pads of the flight animals became thinner. Histological examination of the humerus did not demonstrate significant mineral losses. However, X-ray microtomography showed changes in the trabecular structure in the subepyphyseal zone of bones in flight animals compared to the controls. In summary, all the changes detected in the flight animals were adaptive. Therefore, geckos of this species can be used as an animal model for morphological studies in longer-duration space flights.

Gulimova, V. I.