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Biomedical subjects

M W Luttges

Publications and source records attributed to M W Luttges.

At least 19 recordsLinked to original sources

Low gravity and inertial effects on the growth of E. coli and B. subtilis in semi-solid media.

BACKGROUND: Several published experimental results have shown that cultures of suspended bacteria exhibit increased growth in the spaceflight environment. HYPOTHESIS AND METHODS: To test whether these differences were due to fluid mechanics and not cellular effects, E. coli and B. subtilis were grown on agar cultures under static, agitated, and rotated conditions in the laboratory, and under low-gravity conditions on four Space Shuttle flights. Growth experiments were terminated with glutaraldehyde, and individual cells were counted after quantitative elution from the agar. RESULTS: The spaceflight results, in conjunction with static, rotation, and agitation experiments indicate that E. coli and B. subtilis cultures on agar, unlike their suspension grown counterparts, do not experience heightened final cell concentration when the inertial environment is changed. CONCLUSIONS: This finding points to fluid dynamics and extracellular transport phenomena and not cellular dynamics as the most likely cause of previously reported increases in bacterial growth in microgravity.

Bacillus subtilis↗

Suspension osteopenia in mice: whole body electromagnetic field effects.

Whole-body fields were tested for their efficacy in preventing the osteopenia caused by tail suspension in mice. The fields had fundamental frequencies corresponding to the upper range of predicted endogenous impact-generated frequencies (0.25-2.0 kHz) in the long bones. Three distinct whole-body EMFs were applied for 2 weeks on growing mice. Structural, geometric, and material properties of the femora, tibiae, and humeri of suspended mice were altered compared to controls. Comparison of suspended mice and mice subjected to caloric restriction indicates that the changes in caloric intake do not explain either the suspension or the field-induced effects. In agreement with past studies, rather, unloading appears to cause the suspension effects and to be addressed by the EMFs. The EMF effects on bone properties were apparently frequency dependent, with the lower two fundamental frequencies (260 and 910 Hz) altering, albeit slightly, the suspension-induced bone effects. The fields are not apparently optimized for frequency, etc., with respect to therapeutic potential; however, suspension provides a model system for further study of the in vivo effects of EMFs.

Animals↗

Method for determining individual neuron size in simultaneous single-unit recordings.

A technique for estimating the size of neurons is based on extracellular recordings with paired-electrode sets. Simultaneous single-unit recordings are obtained from the dragonfly mesothoracic ganglion. It is assumed that the ganglion is a passive electrical environment, where spike amplitudes decrease with the inverse of distance squared, and spike angles (widths) increase linearly with distance from the cellular source to the recording electrodes. Starting with the recorded spike amplitudes and angles for each cell, a numerical algorithm is iterated to estimate the true value of the amplitude and angle minus these passive electrical distance effects. The resolved amplitude is a direct, consistent estimate of the size of each recorded neuron. The results indicate that a dichotomy of small and large cells is recorded in roughly a 2:1 ratio. The dichotomy of cell sizes is consistent with the available histological data, although a larger ratio of small to large cells (approximately 10:1) would be expected. Thus, a sampling bias for large cells is apparent, which may be reflective of the larger soma/proximal geometries of such cells. As the technique determines the size of each individual neuron, such biases are eliminated from population studies of the neural tissue. Furthermore, knowledge about the size of each individual neuron permits more detailed analyses of the interactions and contributions of single cells within a network of cells based upon size.

Action Potentials↗

Temperature prediction of space flight experiments by computer thermal analysis.

Life sciences experiments are especially sensitive to temperature. A small temperature difference between otherwise identical samples can cause various differences in biological reaction rates. Knowledge of experimental temperatures and temperature histories help to distinguish the effects of microgravity and temperature on spaceflight experiments compared to ground based studies, and allow appropriate controls and sensitivity tests. Up to the present time, the Orbiter (Space Shuttle) has not generally provided temperature measurement instrumentation inside ambient lockers located in the Mid-deck of the Orbiter, or inside similar facilities such as Spacehab and Spacelab, but many pieces of hardware do have temperature recording capability. Most of these temperatures, however, have only been roughly measured or estimated. Such reported experimental temperatures, while accurate within a range of several degrees Celsius, are of limited utility to biological researchers. The temperature controlled lockers used in spaceflight, such as Commercial-Refrigeration Incubation Modules (C-R/IMs), severely reduce the mass and volume available for test samples and do not necessarily provide uniform thermal environments. While these test carriers avoid some of the experimental temperature variations of the ambient lockers, the number of samples which can be accommodated in these temperature controlled units is limited. In the present work, improved models of thermal prediction and control were sought. Temperatures are predicted by thermal analysis software using empirical temperatures recorded during STS-57. These temperatures are compared to data recorded throughout the mission using Ambient Temperature Recorders (ATRs) located within several payload lockers. Additional test cases are undertaken using controlled ground experiments to more precisely determine the reliability of the thermal model. The approach presented should increase the utility of various spaceflight carriers in the support of biological and material science research and ground control studies done in preparation for flight.

Air Conditioning↗

Effect of microgravity, temperature, and concentration on fibrin and collagen assembly.

In purified form collagen and fibrin can be processed into gel-like matrices of interconnecting fibers. The microscopic structure of materials produced from these macromolecules is critical to their utility as biomaterials. Varying the conditions of the assembly environment allows for the production of a wide range of morphologies. In this study, changes in gravity, temperature, and concentration were examined. Contrary to protein crystal growth studies which indicate substantial increases in organization and size in microgravity, the gravitational environment had no repeatable effect on collagen and fibrin fiber diameters and matrix porosity. However, fibrin gels formed in microgravity appeared more homogeneous than ground samples. Changes in temperature and concentration of both protein and buffer had substantial effects on fiber diameters and material porosity for both collagen and fibrin. Temperature experiments were performed over the range 23.8 to 39 degrees C for fibrin and 22 to 33 degrees C for collagen. Thrombin concentration was varied from 0.02 to 0.10 units/ml for fibrin experiments and buffer concentration was varied by means of a dialysis membrane for collagen experiments. Consequently, the temperature and concentration controls developed for flight experiments are being considered for their potential in developing fibrin and collagen based materials with well-defined microscopic structures. The increased homogeneity of fibrin gels produced in microgravity suggests the possibility of using this environment for the production of optimal biomaterials.

Animals↗

An autonomous module for supporting mice during spaceflight.

The Animal Module for Autonomous space Support (A-MASS) was developed to enable 30-day spaceflight for mice on the first Commercial Experiment Transporter mission. Because space hardware did not previously exist to support mice without astronaut intervention, the A-MASS presented considerable technical and animal care challenges. The technical challenges included maintaining a 42.5l payload volume and 20-g structural conformance while providing 30 days of autonomous mouse support. Sensors, video, a pressurized oxygen supply system and an internal data logging system were incorporated. The A-MASS met NIH guidelines for temperature, humidity, food and water access, oxygen supply, air quality and odor control. These technical and animal care challenges, along with power and mass constraints, were addressed using a novel design which ensures a fresh food and water supply, a clean view path into the cage for the camera system, and removal of the wastes from the air supply. The payload was successfully tested in an enclosed chamber and passed animal health, vibrational, mechanical, and electrical tests. The physiological, tactical and animal support information gathered will be applicable to the development of mouse support modules for the Shuttle Middeck and Space Station Freedom Express Rack environments.

Animal Welfare↗

Contribution of dietary and loading changes to the effects of suspension on mouse femora.

The present study assessed the contributions of feeding changes and unloading to the overall measured effects of 2-wk hindlimb (Tail) suspension on the mouse femora. Feeding changes were addressed by considering the effects of matched feeding among suspended and control mice. The effects of hind limb unloading were considered by comparing suspended mice to mice equipped identically (though not suspended) and matched-fed. The feeding and unloading aspects of suspension appear to cause distinctly differing effects on the stereotypic modeling of the femora. Matched-feeding was accompanied by increased resorption surface in comparison to suspended mice, while unloading led to reduced bone formation at the mid-diaphysis of the femora. Reduced mineral content was observed in the bones of suspended mice when compared to the other mice groups, but without increased resorption surface. Thus, the unloading aspects of the antiorthostatic suspension protocol apparently causes reduced formation and mineralization in the femur.

Animals↗

Class I and class II major histocompatibility molecules play a role in bone marrow-derived macrophage development.

Class I and class II major histocompatibility complex (MHC) molecules play significant roles in T cell development and immune function. We show that MHCI- and MHCII-deficient mice have low numbers of macrophage precursors and circulating monocytes, as well as abnormal bone marrow cell colony-stimulating factor type 1 secretion and bone composition. We suggest that MHCI and MHCII molecules play a significant role in macrophage development.

Animals↗

Plant-module for autonomous space support (P-MASS).

A wide variety of technical and science questions arise when attempting to envision the long-term support of plants, algae and bacteria in space. Currently, spaceflight data remain elusive since there are no U.S. carriers for investigating either the germane technical or scientific issues. The first flight of the Commercial Experiment Transporter (COMET) will provide a nominal 30 day orbital opportunity to evaluate such issues. The P-MASS is a small payload that is designed to meet the mass (40 lbs.), volume (1.5 cu.ft.), and power (120 W) constraints of one of several COMET payloads while enabling flight evaluations of plants, algae and bacteria. Various P-MASS subsystems have been subjected to extensive ground tests as well as KCl35 tests. Various biological sub-systems have been similarly evaluated. Through a variety of sensors coupled with color video, the P-MASS performance and the supported biological systems will be compared for terrestrial controls versus spaceflight materials. This small, low cost payload should return valuable information regarding the requirements for hardware and biological systems needed to move toward bioregenerative life support systems in space. In addition, it should be possible to accurately identify major unresolved difficulties that may arise in the long-term, spaceflight support of various biological systems. Finally, this generic spaceflight capability should enable a variety of plant research programs focused on the use of microgravity to modulate and exploit plant products for commercial applications ranging from new agricultural products to pharmacological feedstocks and new controlled agricultural strategies.

Air Conditioning↗

Four educational programs in Space Life Sciences.

Four different educational programs impacting Space Life Sciences are described: the NASA/USRA Advanced Design Program, the NASA Specialized Center of Research and Training (NSCORT) Program, the Centers for the Commercial Development of Space (CCDS) Program, and the NASA Graduate Research Fellow Program. Each program makes somewhat different demands on the students engaged in them. Each program, at the University of Colorado, involves Space Life Sciences training. While the Graduate Student Research Fellow and NSCORT Programs are discipline oriented, the Advanced Design and CCDS Programs are focused on design, technologies and applications. Clearly, the "training paradigms" differ for these educational endeavors. But, these paradigms can be made to mutually facilitate enthusiasm and motivation. Discipline-oriented academic programs, ideally, must be flexible enough to accommodate the emergent cross-disciplinary needs of Space Life Sciences students. Models for such flexibility and resultant student performance levels are discussed based upon actual academic and professional records.

Biological Science Disciplines↗

Effect of microgravity on collagenase deproteinization and EDTA decalcification of bone fragments.

Undecalcified (n = 140) and decalcified (n = 11) bone fragments were treated with either collagenase (to remove collagen portion; undecalcified n = 64, decalcified n = 11) or EDTA (to remove mineral portion; n = 76) under the reduced gravity environment on US Space Shuttle mission STS-57. The fragments were initially stored in Dulbecco's phosphate buffer solution. After orbit had been established, fragments were exposed to either a neutral buffered collagenase or EDTA solution. Reactions were terminated (neutral buffered formalin for collagenase, 21% CuSO4 5H2O for EDTA) before reentry to earth's atmosphere. Differences in bone samples mass from before flight to after flight were measured. EDTA-treated sample mass was corrected for CuSO4 content. Flight and matched ground (gravitational control) sample showed similar EDTA-induced loss of mineral mass. Collagenase treatments, however, appeared to be more effective in flight samples compared to ground control samples. The flight-exposed, collagenase-treated samples showed significantly more loss of mass than did ground samples. The microgravity environment appeared to promote proteolytic reactions in bone more than the EDTA decalcification reaction.

Animals↗

The role of sex and genotype on antiorthostatic suspension effects on the mouse peripheral skeleton.

Previous antiorthostatic suspension studies have used a single sex and strain of rat or mouse. Nonetheless, broadly similar effects of suspension on the two species indicates a generalized effect of suspension not attributed to specific genetic, behavioral, or sex-linked etiology. In order to directly test genetic and sex-linked factors, the effects of suspension on the appendicular bone of male and female BALB-CJ, C57BL-6J, and DBA-2J mice were compared. These genotypes were selected based on their widely different developmental and behavioral characteristics as well as on past research involving a heterogeneous strain derived from these strains. The effects of suspension on the geometric, mechanical, and material properties of the femora, humeri, and tibiae were determined. Among the bone types, the femora were most significantly affected by suspension. The effects of suspension were similar in nature in male and female mice aged 1.7 months. Strain-dependent suspension effects may be indicative of bone developmental differences in the strains at the age chosen.

Animals↗

Effects of rehydration state on the flexural properties of whole mouse long bones.

The effects of bone water content during dehydration and rehydration on the flexural properties of whole mouse femora were evaluated using three-point bending. The elastic and plastic flexural properties of the bones were determined on a dry mass normalized basis over dehydration times ranging from 0.25 to 48.0 hr; and (following complete dehydration) rehydration times ranging from 0.08 to 12.0 hr. Bones stored in physiological saline for times < 1 hr served as the control group. As expected, dehydration produced increased stiffness and strength along with decreased ductility. Upon rehydration, a statistically significant linear dependence of mechanical properties on recovered free water was obtained for all parameters except the maximum load. Elastic mechanical properties comparable to the controls were regained at differing rates and levels of recovered water content; however, after 3 hr of rehydration there were no statistically significant differences with respect to the control values. The results of this study indicate that the original flexural properties of whole mouse femora are preserved by air dehydration and can be recovered using appropriate saline rehydration intervals.

Animals↗

Skeletal unloading causes organ-specific changes in immune cell responses.

The effects of skeletal unloading using antiorthostatic tail suspension on the mouse immune system are tissue specific. This phenomenon was demonstrated by analyzing cells from the lymph nodes, spleen, and bone marrow. Phytohemagglutinin-induced T-cell proliferation was depressed in lymph nodes after 11 days of antiorthostatic suspension. In contrast, splenic T-cell proliferation in response to phytohemagglutinin was enhanced. Splenic natural killer cell cytotoxicity was unchanged after suspension, which demonstrated the organ- and cell-specific effects of skeletal unloading. Whereas antiorthostatic suspension induced minimal changes in bone, there was a significant depression in the number of macrophage precursors in the bone marrow. Overall, skeletally unloaded animals had slightly higher blood corticosterone levels than did control animals; however, it did not appear to be responsible for the observed changes. In conclusion, skeletal unloading produces organ- and cell-specific changes in the murine immune system rather than a generalized immunosuppression.

Animals↗

Characterization of fluid distribution through a porous substrate under dynamic g conditions.

Dedicated electronic hardware has been constructed to monitor fluid distributions inside a plant rooting/nutrient substrate (Rockwool). With this hardware the effect of dynamically varying gravity states, from enhanced 2g to reduced 0.01g, on solution distributions inside a cube of substrate was monitored aboard the NASA KC-135 reduced gravity research aircraft. The 8 vertices and the center of the cube were used to place sinusoidal voltage sources (electrodes), emitting different fixed frequencies, inside the substrate. Using another set of 9 electrodes the voltage fields were detected across all frequencies. Since the substrate cannot conduct, those frequencies which appeared on any detector (sensor) were indicative of the conductive liquid pathways inside the substrate. An analysis algorithm was developed to visualize the fluid distributions under g-level conditions. Even though the duration of the experiment was short, gravity induced changes in fluid position were readily and reliably detected. Since fluids carry the nutrients necessary for plant growth these data and techniques can lead to the development of a uniform nutrient supply system supportive of optimal plant growth in space.

Algorithms↗

The physical and mechanical effects of suspension-induced osteopenia on mouse long bones.

The present investigation addresses the extent of tail-suspension effects on the long bones of mice. The effects are explored in both sexes, in both forelimb and hindlimb bones, and in both diaphyseal and metaphyseal/epiphyseal bones. Two weeks of suspension provided unloading of the femora and tibiae and an altered loading of the humeri. Whole-bone effects included lower mass (approximately 10%) and length (approximately 4%) in the bones of suspended mice compared to controls. The geometric and material properties of the femora were considered along the entire length of the diaphysis and in the metaphysis/epiphysis portions as a unit. Geometric effects included lower cross-sectional cortical area (16%), cortical thickness (25%) and moment of inertia (21%) in the femora of suspended mice; these differences were observed in both distal and proximal portions of the femur diaphysis. The relative amount of bone comprising the middle 8 mm of the diaphysis was greater (3%) in the control mice than in the suspended mice. Significant mass differences between the group in the metaphysis/epiphysis were not observed. Material effects included lower %ash (approximately 2%) in the femora and tibiae as well as in the humeri of suspended mice compared to controls. With respect to the measured physical and material properties, suspension produced similar bone responses in male and female mice. The effects of suspension are manifested largely through geometric rather than through material changes.

Animals↗

Effect of oral calcium and calcium + fluoride treatments on mouse bone properties during suspension.

The bone effects of oral dosages of calcium chloride with or without supplementary sodium fluoride were assessed in antiorthostatically suspended mice. Two calcium dosages were used to replace half (3.1 mM) or all(6.3 mM) of the dietary calcium lost due to reduced food intake by the suspended mice. Two groups of 6.3 mM CaCl2-treated mice were additionally treated with 0.25 or 2.5 mM NaF. The results indicate that supplementation of the mouse drinking water with calcium salts prevents bone changes induced by short-term suspension, while calcium salts in combination with fluoride are less effective as fluoride dosage increases. However, the calcium supplements change the relationship between the femur mechanical properties and the mineral composition of the bone. Because of this, it appears that oral calcium supplements are effective through a mechanism other than simple dietary supplementation and may indicate a dependence of bone consistency on systemic and local fluid conditions.

Animals↗

Recognizing and optimizing flight opportunities with hardware and life sciences limitations.

The availability of orbital space flight opportunities to conduct life sciences research has been limited. It is possible to use parabolic flight and sounding rocket programs to conduct some kinds of experiments during short episodes (seconds to minutes) of reduced gravity, but there are constraints and limitations to these programs. Orbital flight opportunities are major undertakings, and the potential science achievable is often a function of the flight hardware available. A variety of generic types of flight hardware have been developed and tested, and show great promise for use during NSTS flights. One such payload configuration is described which has already flown.

Aircraft↗