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NASA NTRS2012

Mass and Momentum Transport in Microcavities for Diffusion-Dominant Cell Culture Applications

For the informed design of microfluidic devices, it is important to understand transport phenomena at the microscale. This letter outlines an analytically-driven approach to the design of rectangular microcavities extending perpendicular to a perfusion microchannel for microfluidic cell culture devices. We present equations to estimate the spatial transition from advection- to diffusion-dominant transport inside cavities as a function of the geometry and flow conditions. We also estimate the time required for molecules, such as nutrients or drugs to travel from the microchannel to a given depth into the cavity. These analytical predictions can facilitate the rational design of microfluidic devices to optimize and maintain long-term, physiologically-based culture conditions with low fluid shear stress.

Yew, Alvin G.↗

NASA NTRSDate not supplied

AMMPER: Agent-based Model for Microbial Populations Exposed to Radiation

Exposure of microbial populations to galactic cosmic radiation (GCR) in the deep-space radiation environment may lead to intracellular damage that compromises the ability of cells to repair and replicate. While simulation programs of deep-space radiation do exist, they focus primarily on single-cell damage, rather than population-wide effects. In this work, a new application and graphical user interface, Agent-Based Model for Microbial Populations Exposed to Radiation (AMMPER) is presented, which simulates the effects of proton-based GCR on Saccharomyces cerevisiae population growth. AMMPER consists of a 50x50x50 µm simulation space, analogous to an aqueous culture medium with non-limiting nutrient and pH buffering, in a microwell plate or microfluidic culture card. This model utilizes Relativistic Ion Tracks (RITRACKS) to create detailed track structures of the radiation traversals. AMMPER then calculates the radiation dose present at each cell, and subsequently determines the damage (chromosomal aberrations, oxidative stress, etc.) and resulting loss of cell viability from both primary and secondary radiative effects. Through implementing cell replication, repair, damage, and death, the effect of radiation exposure on the population growth can be determined. With AMMPER, long-duration effects of the deep space environment on entire populations can be determined and used to assess the feasibility of sustaining life in space.

Amrita Singh↗

NASA NTRS2018

Global Patterns of Crop Yield Stability Under Additional Nutrient and Water Inputs

Agricultural production must increase to feed a growing and wealthier population, as well as to satisfy increasing demands for biomaterials and biomass-based energy. At the same time, deforestation and land-use change need to be minimized in order to preserve biodiversity and maintain carbon stores in vegetation and soils. Consequently, agricultural land use needs to be intensified in order to increase food production per unit area of land. Here we use simulations of AgMIP's Global Gridded Crop Model Intercomparison (GGCMI) phase 1 to assess implications of input-driven intensification (water, nutrients) on crop yield and yield stability, which is an important aspect in food security. We find region- and crop-specific responses for the simulated period 1980+/-2009 with broadly increasing yield variability under additional nitrogen inputs and stabilizing yields under additional water inputs (irrigation), reflecting current patterns of water and nutrient limitation. The different models of the GGCMI ensemble show similar response patterns, but model differences warrant further research on management assumptions, such as variety selection and soil management, and inputs as well as on model implementation of different soil and plant processes, such as on heat stress, and parameters. Higher variability in crop productivity under higher fertilizer input will require adequate buffer mechanisms in trade and distribution/storage networks to avoid food price volatility.

Maize↗

NASA NTRSDate not supplied

To Cage or Not to Cage Seagrass; Is There Even a Question?

To Cage or Not to Cage Seagrass; Is There Even a Question? Loraé T. Simpson1, Morgan K. Gilligan1, Douglas M. Scheidt2 1Florida Oceanographic Society, Stuart, FL; 2Herndon Solutions Group/NEMCON, Kennedy Space Center, FL Contact email: lsimpson@floridaocean.org Diminished water quality has led to the decline of seagrasses and their associated ecosystem services in the Banana River Lagoon (BRL), resulting in increased restoration initiatives. Restoration success may be hindered by herbivory and the addition of herbivore exclusion devices may limit grazing stress on newly established seagrasses. These structures may alter sediment dynamics, influencing seagrass productivity. To test whether herbivore exclusion devices impact restoration success and sediment dynamics, we caged half of our seagrass restoration plots in the BRL. Treatments were monitored monthly for seagrass growth, and at three months for porewater nutrients, sediment bulk density, and carbon to nitrogen ratio. On average, caged plots had 87% more seagrass cover and 81% taller blades than control plots, highlighting the importance of caging newly transplanted seagrasses. There was no significant difference in porewater or sediment characteristics between treatments, suggesting that pedogensis and nutrient cycling need extended monitoring to capture changes in temporal scales.

Indian River Lagoon↗

NASA NTRSDate not supplied

To Cage or Not to Cage Seagrass; Is There Even a Question?

Diminished water quality has led to the decline of seagrasses and their associated ecosystem services in the Banana River Lagoon (BRL), resulting in increased restoration initiatives. Restoration success may be hindered by herbivory and the addition of herbivore exclusion devices may limit grazing stress on newly established seagrasses. These structures may alter sediment dynamics, influencing seagrass productivity. To test whether herbivore exclusion devices impact restoration success and sediment dynamics, we caged half of our seagrass restoration plots in the BRL. Treatments were monitored monthly for seagrass growth, and at three months for porewater nutrients, sediment bulk density, and carbon to nitrogen ratio. On average, caged plots had 87% more seagrass cover and 81% taller blades than control plots, highlighting the importance of caging newly transplanted seagrasses. There was no significant difference in porewater or sediment characteristics between treatments, suggesting that pedogensis and nutrient cycling need extended monitoring to capture changes in temporal scales.

Indian River Lagoon↗

NASA NTRS1988

Human cell culture in a space bioreactor

Microgravity offers new ways of handling fluids, gases, and growing mammalian cells in efficient suspension cultures. In 1976 bioreactor engineers designed a system using a cylindrical reactor vessel in which the cells and medium are slowly mixed. The reaction chamber is interchangeable and can be used for several types of cell cultures. NASA has methodically developed unique suspension type cell and recovery apparatus culture systems for bioprocess technology experiments and production of biological products in microgravity. The first Space Bioreactor was designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small Bioreactor is being constructed for flight experiments in the Shuttle Middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption and control of low shear stress on cells.

Morrison, Dennis R.↗

NASA NTRS1987

Suspension cell culture in microgravity and development of a space bioreactor

NASA has methodically developed unique suspension type cell and recovery apparatus culture systems for bioprocess technology experiments and production of biological products in microgravity. The first space bioreactor has been designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small (500 ml) bioreactor is being constructed for flight experiments in the Shuttle middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption, and control of low shear stress on cells.

Morrison, Dennis R.↗

NASA NTRS1994

Dynamics of microorganism populations in recirculating nutrient solutions

This overview covers the basic microbial ecology of recirculating hydroponic solutions. Examples from NASA and Soviet Controlled Ecological Life Support Systems (CELSS) tests and the commercial hydroponic industry will be used. The sources of microorganisms in nutrient solutions include air, water, seeds, plant containers and plumbing, biological vectors, and personnel. Microbial fates include growth, death, and emigration. Important microbial habitats within nutrient delivery systems are root surfaces, hardware surfaces (biofilms), and solution suspension. Numbers of bacteria on root surfaces usually exceed those from the other habitats by several orders of magnitude. Gram negative bacteria dominate the microflora with fungal counts usually much lower. Trends typically show a decrease in counts with increasing time unless stressed plants increase root exudates. Important microbial activities include carbon mineralization and nitrogen transformations. Important detrimental interactions include competition with plants, and human and plant pathogenesis.

Strayer, R. F.↗

NASA NTRS1994

Dynamics of microorganism populations in recirculating nutrient solutions

This overview covers the basic microbial ecology of recirculating hydroponic solutions. Examples from NASA and Soviet CELSS tests and the commercial hydroponic industry will be used. The sources of microorganisms in nutrient solutions include air, water, seeds, plant containers and plumbing, biological vectors, and personnel. Microbial fates include growth, death, and emigration. Important microbial habitats within nutrient delivery systems are root surfaces, hardware surfaces (biofilms), and solution suspension. Numbers of bacteria on root surfaces usually exceed those from the other habitats by several orders of magnitude. Gram negative bacteria dominate the microflora with fungal counts usually much lower. Trends typically show a decrease in counts with increasing time unless stressed plants increase root exudates. Important microbial activities include carbon mineralization and nitrogen transformations. Important detrimental interactions include competition with plants, and human and plant pathogenesis.

Strayer, R. F.↗

NASA NTRS2014

Estimating Major Ion and Nutrient Concentrations in Mangrove Estuaries in Everglades National Park Using Leaf and Satellite Reflectance

Coastal mangrove ecosystems are under duress worldwide because of urban development, sea-level rise, and climate change, processes that are capable of changing the salinity and nutrient concentration of the water utilized by the mangroves. This study correlates long-term water chemistry in mangrove environments, located in Everglades National Park,with mangrove spectral reflectance measurements made at both the leaf and canopy scales. Spectral reflectance measurements were collected using a handheld spectrometer for leaf-level measurements and Landsat 5TMdata for regional coverage. Leaf-level reflectance data were collected from three mangrove species (i.e., red, black and white mangroves) across two regions; a tall mangrove (approximately 18m) and dwarf mangrove (1- 2 m) region. The reflectance data were then used to calculate a wide variety of biophysical reflectance indices (e.g., NDVI, EVI, SAVI) to determine signs of stress. Discrete, quarterly water samples from the surface water, groundwater, and pore water (20 and 85 cm depths) and daily autonomous surface water samples were collected at each site and analyzed for major anions (Cl(−) and SO(2(-)/4)), cations (Na(+), K(+), Mg(sup 2(+)), and Ca(sup 2(+)), total nitrogen (TN) and total phosphorus (TP).Mangrove sites that exhibited the highest salinity and ionic concentrations in the surface and subsurface water also had the lowest near-infrared reflectance at both the leaf and satellite levels. Seasonal reflectance responses were measured in the near-infrared (NIR) wavelengths at both the leaf and canopy scales and were strongly correlated with nutrient and ionic concentrations in the surface and subsurface water, even though there was no significant separability between the three mangrove species. Study sites that experienced the greatest variability in surface and subsurface water ionic concentrations also exhibited the greatest fluctuations in NIR spectral reflectance. Landsat 5TM images were able to detect tall and dwarf mangroves by the differences in spectral indices (e.g., NDVI, NDWI, and EVI) because of the variability in the background conditions amongst the environments. In addition, Landsat 5TM images spanning 16 years (1993- 2009) were successfully used to estimate the seasonal variability in ionic concentrations in the surface water across the Florida Coastal mangrove ecotone. This study has shown that water chemistry can be estimated indirectly by measuring the change in spectral response at the leaf- or satellite-scale. Furthermore, the results of this research may be extrapolated to similar coastal mangrove systems throughout the Caribbean and world-wide wherever red, black, and white mangroves occur.

Mangroves↗

NASA NTRS1989

Comparison of Soviet and US space food and nutrition programs

The Soviet Space Food and Nutrition programs are compared with those of the U.S. The Soviets established the first Space Food programs in 1961, when one of the Soviet Cosmonauts experienced eating in zero gravity. This study indicates that some major differences exist between the two space food and nutrition programs regarding dietary habits. The major differences are in recommended nutrient intake and dietary patterns between the cosmonauts and astronauts. The intake of protein, carbohydrates and fats are significantly higher in cosmonaut diets compared to astronauts. Certain mineral elements such as phosphorus, sodium and iron are also significantly higher in the cosmonauts' diets. Cosmonauts also experience intake of certain unconventional food and plant extracts to resist stress and increase stamina.

Ahmed, Selina↗

NASA NTRS1992

Closed Ecological Life Support Systems (CELSS) Test Facility

The CELSS Test Facility (CTF) is being developed for installation on Space Station Freedom (SSF) in August 1999. It is designed to conduct experiments that will determine the effects of microgravity on the productivity of higher (crop) plants. The CTF will occupy two standard SSF racks and will accommodate approximately one square meter of growing area and a canopy height of 80 cm. The growth volume will be isolated from the external environment, allowing stringent control of environmental conditions. Temperature, humidity, oxygen, carbon dioxide, and light levels will all be closely controlled to prescribed set points and monitored. This level of environmental control is needed to prevent stress and allow accurate assessment of microgravity effect (10-3 to 10-6 x g). Photosynthetic rates and respiration rates, calculated through continuous recording of gas concentrations, transpiration, and total and edible biomass produced will be measured. Toxic byproducts will be monitored and scrubbed. Transpiration water will be collected within the chamber and recycled into the nutrient solution. A wide variety of crop plants, e.g., wheat, soy beans, lettuce, potatoes, can be accommodated and various nutrient delivery systems and light delivery systems will be available. In the course of its development, the CTF will exploit fully, and contribute importantly, to the state-of-art in closed system technology and plant physiology.

Macelroy, Robert D.↗

NASA NTRS2011

Hydrofocusing Bioreactor Produces Anti-Cancer Alkaloids

A methodology for growing three-dimensional plant tissue models in a hydrodynamic focusing bioreactor (HFB) has been developed. The methodology is expected to be widely applicable, both on Earth and in outer space, as a means of growing plant cells and aggregates thereof under controlled conditions for diverse purposes, including research on effects of gravitation and other environmental factors upon plant growth and utilization of plant tissue cultures to produce drugs in quantities greater and at costs lower than those of conventional methodologies. The HFB was described in Hydro focus - ing Bioreactor for Three-Dimensional Cell Culture (MSC-22358), NASA Tech Briefs, Vol. 27, No. 3 (March 2003), page 66. To recapitulate: The HFB offers a unique hydrofocusing capability that enables the creation of a low-shear liquid culture environment simultaneously with the herding of suspended cells and tissue assemblies and removal of unwanted air bubbles. The HFB includes a rotating cell-culture vessel with a centrally located sampling port and an internal rotating viscous spinner attached to a rotating base. The vessel and viscous spinner can be made to rotate at the same speed and direction or different speeds and directions to tailor the flow field and the associated hydrodynamic forces in the vessel in order to obtain low-shear suspension of cells and control of the locations of cells and air bubbles. For research and pharmaceutical-production applications, the HFB offers two major benefits: low shear stress, which promotes the assembly of cells into tissue-like three-dimensional constructs; and randomization of gravitational vectors relative to cells, which affects production of medicinal compounds. Presumably, apposition of plant cells in the absence of shear forces promotes cell-cell contacts, cell aggregation, and cell differentiation. Only gentle mixing is necessary for distributing nutrients and oxygen. It has been postulated that inasmuch as cells in the simulated microgravitation of an HFB do not need to maintain the same surface forces as in normal Earth gravitation, they can divert more energy sources to growth and differentiation and, perhaps, to biosynthesis of greater quantities of desired medicinal compounds. Because one can adjust the HFB to vary effective gravitation, one can also test the effects of intermediate levels of gravitation on biosynthesis of various products. The potential utility of this methodology for producing drugs was demonstrated in experiments in which sandalwood and Madagascar periwinkle cells were grown in an HFB. The conditions in the HFB were chosen to induce the cells to form into aggregate cultures that produced anti-cancer indole alkaloids in amounts greater than do comparable numbers of cells of the same species cultured according to previously known methodologies. The observations made in these experiments were interpreted as suggesting that the aggregation of the cells might be responsible for the enhancement of production of alkaloids.

Gonda, Steve R.↗

NASA NTRSDate not supplied

Spaceflight Food System Impacts to Nutritional Adequacy, Health, Performance, and Resources in Space Exploration

Despite high physical standards and training protocols, physiological and behavioral decrements have been documented in astronauts on both short (1-2 weeks) and long (6+ month) missions in spaceflight, including dysregulation of the immune system, cardiovascular and musculoskeletal deconditioning, ophthalmic changes, weight loss, and increased stress and fatigue. Optimizing food and nutrition intakes are key underpinnings for the proper function and performance of all physiological systems and the resulting physical and behavioral health and performance outcomes of astronauts. Much has been learned about the role of nutrition in human health on Earth over the past hundred years, from the identity and role of specific vitamins to the importance of the quantities of some nutrients to immune function. The requirements for providing adequate nutrition to astronauts seem obvious. However, providing a safe, reliable, and nutritious food system for space exploration missions remains a challenge. In fact, food is one of the greatest resource and logistical challenges, which is part of why it remains a “red” risk for Mars missions.

Grace L Douglas↗

NASA NTRSDate not supplied

Space Algae-2: Preflight Testing for A Long-Duration, Multi-Omics Analysis of Arthrospira Platensis

The cyanobacteria Arthrospira platensis NIES-39, commonly known as spirulina, could provide a fresh supply of nutrients for crew on long-duration spaceflight missions. Spirulina is a readily digestible food that is high in protein with all essential amino acids as well as significant levels of B vitamins, antioxidants, and anti-inflammatory metabolites. Spaceflight has multiple abiotic stressors such as increased ionizing radiation and microgravity, which causes a lack of convective mixing. These environmental conditions may impact productivity, nutritional composition, and in long-duration propagation, spaceflight stress may impact the genetic stability of spirulina cultures. We are developing an International Space Station experiment to continuously culture A. platensis for six months. Multi-omics profiling will be used to monitor for changes in the genome, transcriptome, proteome, and metabolome to determine if A. platensis is a suitable nutritional supplement on long-duration missions. During preflight testing we developed a protocol for inoculated liquid cultures to survive a 10-week storage period prior to photo-incubation. The bioreactor bag, temperature, and lighting conditions that support a 14-day growth cycle between passages were also determined. Media testing identified minimal salts supporting robust growth that can be stored in liquid or dry form. A simple filtration method was developed to dewater cultures and harvest biomass for frozen sample return. We optimized a cryopreservation method to enable return of live cells for isolation of individual A. platensis clones. The concept of operations for Space Algae-2 developed from these test results as well as progress on multi-omics analysis methods will be presented.

Algae↗

NASA NTRSDate not supplied

Space Algae-2: Preflight Testing for A Long-Duration, Multi-Omics Analysis of Arthrospira Platensis

Algae↗

NASA NTRS2001

Evidence for a role in growth and salt resistance of a plasma membrane H+-ATPase in the root endodermis

The plasma membrane of plant cells is energized by an electrochemical gradient produced by P-type H+-ATPases (proton pumps). These pumps are encoded by at least 12 genes in Arabidopsis. Here we provide evidence that isoform AHA4 contributes to solute transport through the root endodermis. AHA4 is expressed most strongly in the root endodermis and flowers, as suggested by promoter-GUS reporter assays. A disruption of this pump (aha4-1) was identified as a T-DNA insertion in the middle of the gene (after VFP(574)). Truncated aha4-1 transcripts accumulate to approximately 50% of the level observed for AHA4 mRNA in wild-type plants. Plants homozygous for aha4-1 (-/-) show a subtle reduction in root and shoot growth compared with wild-type plants when grown under normal conditions. However, a mutant phenotype is very clear in plants grown under salt stress (e.g., 75 or 110 mM NaCl). In leaves of mutant plants subjected to Na stress, the ratio of Na to K increased 4-5-fold. Interestingly, the aha4-1 mutation appears to be semidominant and was only partially complemented by the introduction of additional wild-type copies of AHA4. These results are consistent with the hypothesis that aha4-1 may produce a dominant negative protein or RNA that partially disrupts the activity of other pumps or functions in the root endodermal tissue, thereby compromising the function of this cell layer in controlling ion homeostasis and nutrient transport.

NASA Discipline Plant Biology↗

NASA NTRS1989

Control of water and nutrients using a porous tube - A method for growing plants in space

A plant nutrient delivery system that uses a microporous, hydrophilic tube was developed with potential application for crop production in the microgravity of space. The tube contains a nutrient solution and delivers it to the roots. Pumps attached to the tubing create a very small suction that holds the solution within the tube. This system was used to grow wheat for 107 d in a controlled environment at suctions of 0.40, 1.48, or 2.58 kPa. The water absorbed through the pores of the tube by baby diaper sections decreased as suction increased. Correspondingly, final plant biomass, seed number, and spikelet number also tended to decrease as suction increased. The reduced yield at higher suction suggests that the plants experienced water stress, although all suctions were below those typical of soils at field capacity.

Dreschel, Thomas W.↗