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At least 37 records · Page 2
All sourcesNASA NTRSDOE OSTI
NASA NTRS2017

Measuring N2 Pressure Using Cyanobacteria Discipline: Geomicrobiology

The evolution of Earth's atmosphere has been governed by biological evolution. Dinitrogen (N2) has been a major constituent of Earth's atmosphere throughout the planet's history, yet only a few constraints exist for the partial pressure of N2 (pN2). In this study we evaluate two new potential proxies for pN2: the physical spacing between heterocysts and the isotopic signature of nitrogen fixation in filamentous cyanobacteria. Heterocyst-forming filamentous cyanobacteria are some of the oldest photosynthetic microorganisms on Earth, and debated fossilized specimens have been found in sedimentary rocks as old as 2 Ga. These organisms overcome nitrogen limitation in their aqueous environment through cellular differentiation along their filaments. The specialized cells that develop, known as heterocysts, fix the nitrogen and laterally distribute it to neighboring cells along the filaments. Because the concentration of the dissolved N2 available to the filaments correlates directly with pN2, any preservable physiological response of the organism to the changed N2 availability constitutes a potential proxy for pN2. In the laboratory, we have examined how pN2 is reflected in the heterocyst spacing pattern and in the isotopic signature of nitrogen fixation by subjecting the representative species Anabaena cylindrica and Anabaena variabilis to different N2 partial pressures during growth at constant temperature and lighting (in media free of combined nitrogen). We show experimentally that the distance between heterocysts and the nitrogen isotope fractionation measured in bulk biomass reflect the pN2 experienced by Anabaena cylindrica. Current work is investigating these responses in Anabaena variabilis. When heterocystous cyanobacteria fossilize, these morphological and isotopic signatures should preserve information about pN2 at that time. Application of this relationship to the rock record may provide a paleoproxy to complement the two existing geobarometers.

nitrogen↗
DOE OSTI2026

From photosynthetic electron flow to gene regulation: redox signal transduction in cyanobacteria

In cyanobacteria, the free-living ancestors of chloroplasts, photosynthesis simultaneously sustains growth and generates reactive oxygen species (ROS) that damage proteins, lipids, and DNA when light capture outpaces carbon fixation. Maintaining redox balance, therefore, requires cells to read photosynthetic electron flow as a signal that continuously tunes gene expression and protein activity. This review traces how these redox signals are transduced to transcription machinery through three routes: membrane-localized sensors, cytoplasmic redox sensors downstream of photosystem I, and ROS generated when electron sinks are saturated. Membrane-bound histidine kinases (two-component systems) relay the redox state of the plastoquinone pool to control photosystem remodeling, pigment biosynthesis, and circadian timing. Cytoplasmic one-component regulators, by contrast, sense redox directly through thiol-disulfide switches, glutathionylation, iron-sulfur clusters, and metal-catalyzed oxidation to control photosystem-cofactor, electron-carrier, and transition-metal homeostasis. Because many of these regulators persist in algal and plant chloroplasts, cyanobacteria illuminate principles of redox control across photosynthetic eukaryotes. Post-transcriptional and translational control further shapes redox-dependent gene expression programs through transcript stability, ribosome assembly, and translation initiation, extending redox regulation beyond transcription to every step of protein synthesis and even activity modulation. Finally, we connect redox regulation to photosynthetic physiology, stress resilience, and the rational engineering of cyanobacteria for sustainable bioproduction.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Spatial proteomics reveals signal sequence characteristics correlated with localization in cyanobacteria

Abstract Cyanobacteria have an inner and outer cell membrane enclosing the periplasm and cell wall and an additional set of internal membranes (called the thylakoid membranes) enclosing the thylakoid lumen. The periplasm and thylakoid lumen have unique proteomes, but the mechanisms regulating protein sorting to these locations have remained elusive. Here, proximity-based proteomics using the engineered peroxidase APEX2 was performed in the cyanobacteria Synechococcus sp. PCC 7002 to profile the proteomes of the cytoplasm, thylakoid lumen, and the periplasm and outer membrane (P-OM). Our analyses revealed specific roles for the thylakoid lumen in photosynthesis and energy generation, as well as roles for the periplasm in metabolite transport and binding, cell motility, and cell wall maintenance. Forty proteins localized to both the thylakoid lumen and the P-OM; however, their biological functions remain unclear. We also analyzed the correlation between signal sequence characteristics and differential protein localization to either the thylakoid lumen or the P-OM. In PCC 7002, as well as Synechocystis sp. PCC 6803 and Nostoc sp. PCC 7120, thylakoid lumen proteins translocated across membranes via the Secretory (Sec) system possessed more hydrophobic and alpha-helical signal sequence H-regions than P-OM proteins. The signal sequences of homologous proteins in Gloeobacter violaceus PCC 7421, a cyanobacterial species with a combined thylakoid lumen and periplasmic space, did not exhibit such differences. Therefore, the pattern of increased H-region hydrophobicity and alpha helix content is specific to cyanobacteria with a separate thylakoid lumen space and likely contributes to proper protein sorting between the thylakoid lumen and periplasm.

Plant Sciences↗
NASA NTRS2008

Diversity and Physiology of Siderophilic Cyanobacteria: Implication for the Bioenergetics

Prior to 2.4 Ga, global oceans were likely significantly enriched in soluble iron (Rouxel, Bekker, Edwards, 2005), a condition that is not conducive to the growth of most contemporary mesophilic cyanobacteria (CB). Recent studies of the mechanisms of iron-deficiency stress in CB suggest that contemporary mesophilic freshwater and marine B underwent long-term adaptation to a permanent decrease in soluble iron in the ocean environment (Boyer, et al., 1987; Braun, Hantke, and Koster, 1998). Of all extant environments, iron-depositing hot springs may constitute the most appropriate natural models for analysis of the transition of ancestral cyanobacteria (CB) or protocyanobacteria (PCB) (Olson, 2001) from anoxygenic photosynthesis to oxygenic one and biogeochemical processes in the late Archean and early Paleoproterozoic eras. In particular, Olson (2001) proposed the definition for PCB and postulated that the common ancestor of PCB and CB might well have used Fe(OH)+ as the principal electron donor for CO2 fixation (Widdel, et al., 1993; Ehrenreich and Widdel, 1994; Pierson and Olson, 1989; Olson, 2006). Olson (2001) proposed that the driving force for the evolution of RC2, in addition to RC1, was the necessity to use Fe(OH)+ effectively for CO2 fixation in the absence of reduced sulfur compounds. The global decrease of dissolved environmental reduced iron could have been the driving force for the transition from anoxygenic to oxygenic photosynthesis (Brown et al., 2007). Despite the insights into the ecology, evolutionary biology, paleogeobiochemistry, and astrobiology the examination of iron depositing hot springs (IDHS) could potentially provide, very few studies dedicated to the diversity and physiology of cyanobacteria inhabiting IDHS have been conducted. Here we describe the phylogeny, physiology and ultrastructure and biogeochemical activity of several recent CB isolates from two different greater Yellowstone area IDHS, e.g. LaDuke and Chocolate Pots. Phylogenetic analysis of 16S rRNA genes indicated that 6 of 12 new isolates examined could not be placed within established CB genera. Some of the isolates exhibited pronounced requirements for elevated iron concentrations, with maximum growth rates observed when 0.4-1 mM Fe(3+) was present in the media. However, the pronounced effect of iron limitation on the proliferation of siderophilic CB can be observed only after several passages through iron "free" media. TEM studies of several species of siderophilic CB revealed that the cultures JSC-3 and -11 are probably capable of some sort of pinocytosis of precipitated iron. This phenomenon may explain high tolerance of siderophilic CB to iron deficit. We also found that the stimulation of the growth of siderophilic CB by oxidized iron is accompanied by the decrease of O2 evolution by some species after addition Fe(2+) in iron "free" medium.

Brown, Igor↗
NASA NTRS2006

Iron Tolerant Cyanobacteria as an Effective Tool to Study Early Evolution of Life and the Development of Biosignatures

We are currently conducting preliminary studies on the diversity of iron-tolerant cyanobacteria (CB) isolated from iron-depositing hot springs in and around Yellowstone National Park (WY, USA). In conclusion, there is no consensus on the divergence of cyanobacteria from a common ancestor for either anoxygenic or oxygenic phototrophs. Anoxygenic photosynthesis may have provided energy for the common ancestor, but it is unclear what environmental pressure induced the evolving of oxygenic phototrophs. It is supposed, however, that predecessors of contemporary CB were capable of oxidizing various substrates other than water , and it is likely that Fe2+ could be one of those substrates . If that were the case, the work of entire photosystems in Precambrian cyanobacteria and/or in their predecessors could follow three scenarios (at least): 1) ferrous iron may have been oxidized in PS II but without significant effects on oxygen evolution, and environmental iron could have been oxidized either enzymatically or chemically; 2) ferrous iron may have been oxidized only enzymatically by PS II, accompanied by the repression of O2 evolution; or 3) ferrous iron may have been oxidized by PS I upon the prevalence of anoxygenic photosynthesis or without any effect on PS II. All of these scenarios will be the subject of our future studies with the aim to understand which line-ages of CB could be typical for Precambrian time.

Brown, Igor↗
NASA NTRS2015

Genetically Engineered Cyanobacteria

The disclosed embodiments provide cyanobacteria spp. that have been genetically engineered to have increased production of carbon-based products of interest. These genetically engineered hosts efficiently convert carbon dioxide and light into carbon-based products of interest such as long chained hydrocarbons. Several constructs containing polynucleotides encoding enzymes active in the metabolic pathways of cyanobacteria are disclosed. In many instances, the cyanobacteria strains have been further genetically modified to optimize production of the carbon-based products of interest. The optimization includes both up-regulation and down-regulation of particular genes.

Zhou, Ruanbao↗
DOE OSTI2024

Cyanobacteria newly isolated from marine volcanic seeps display rapid sinking and robust, high-density growth

Cyanobacteria are photosynthetic organisms that play important roles in carbon cycling and are promising bioproduction chassis. Here, we isolate two novel cyanobacteria with 4.6Mbp genomes, UTEX 3221 and UTEX 3222, from a unique marine environment with naturally elevated CO₂. We describe complete genome sequences for both isolates and, focusing on UTEX 3222 due to its planktonic growth in liquid, characterize biotechnologically relevant growth and biomass characteristics. UTEX 3222 outpaces other fast-growing model strains on a solid medium. It can double every 2.35 hours in a liquid medium and grows to high density (>31 g/L biomass dry weight) in batch culture, nearly double that of Synechococcus sp. PCC 11901, whose high-density growth was recently reported. In addition, UTEX 3222 sinks readily, settling more quickly than other fast-growing strains, suggesting favorable economics of harvesting UTEX 3222 biomass. These traits may make UTEX 3222 a compelling choice for marine carbon dioxide removal (CDR) and photosynthetic bioproduction from CO₂. Overall, we find that bio-prospecting in environments with naturally elevated CO₂ may uncover novel CO₂-metabolizing organisms with unique characteristics.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1995

Mesoproterozoic Archaeoellipsoides: akinetes of heterocystous cyanobacteria

The genus Archaeoellipsoides Horodyski & Donaldson comprises large (up to 135 micrometers long) ellipsoidal and rod-shaped microfossils commonly found in silicified peritidal carbonates of Mesoproterozoic age. Based on morphometric and sedimentary comparisons with the akinetes of modern bloom-forming Anabaena species, Archaeoellipsoides is interpreted as the fossilized remains of akinetes produced by planktic heterocystous cyanobacteria. These fossils set a minimum date for the evolution of derived cyanobacteria capable of marked cell differentiation, and they corroborate geochemical evidence indicating that atmospheric oxygen levels were well above 1% of present day levels 1,500 million years ago.

NASA Discipline Exobiology↗
NASA NTRS2016

Water Sources for Cyanobacteria Below Desert Rocks in the Negev Desert Determined by Conductivity

We present year round meteorological and conductivity measurements of colonized hypolithic rocks in the Arava Valley, Negev Desert, Israel. The data indicate that while dew is common in the Negev it is not an important source of moisture for hypolithic organisms at this site. The dominance of cyanobacteria in the hypolithic community are consistent with predictions that cyanobacteria are confined to habitats supplied by rain. To monitor the presence of liquid water under the small Negev rocks we developed and tested a simple field conductivity system based on two wires placed about 0.5 cm apart. Based on 21 replicates recorded for one year in the Negev we conclude that in natural rains (0.25 mm to 6 mm) the variability between sensor readings is between 20 and 60% decreasing with increasing rain amount. We conclude that the simple small electrical conductivity system described here can be used effectively to monitor liquid water levels in lithic habitats. However, the natural variability of these sensors indicates that several replicates should be deployed. The results and method presented have use in arid desert reclamation programs.

hypoliths↗
NASA NTRS2018

Mobile Device Application for Monitoring Cyanobacteria Harmful Algal Blooms Using Sentinel-3 Satellite Ocean and Land Colour Instruments

Cyanobacterial harmful algal blooms (cyanoHAB) cause human and ecological health problems in lakes worldwide. The timely distribution of satellite-derived cyanoHAB data is necessary for adaptive water quality management and for targeted deployment of water quality monitoring resources. Software platforms that permit timely, useful, and cost-effective delivery of information from satellites are required to help managers respond to cyanoHABs. The Cyanobacteria Assessment Network (CyAN) mobile device application (app) uses data from the European Space Agency Copernicus Sentinel-3 satellite Ocean and Land Colour Instrument (OLCI) in near real-time to make initial water quality assessments and quickly alert managers to potential problems and emerging threats related to cyanobacteria. App functionality and satellite data were validated with 25 state health advisories issued in 2017. The CyAN app provides water quality managers with a user-friendly platform that reduces the complexities associated with accessing satellite data to allow fast, efficient, initial assessments across lakes.

Harmful Algal Bloom↗
NASA NTRS2023

Spatial and Temporal Characterization of Cyanobacteria Blooms in the Mississippi Sound and Their Relationship to the Bonnet Carré Spillway Openings

During the spring and summer of 2019, an unprecedented cyanobacterial harmful algal bloom (cyanoHAB) was responsible for beach advisories on 25 beaches along the Mississippi Sound for over 3 months. Due to the preceding heavy rainfall and flooding within the Mississippi River watershed, for the first time in history, the Bonnet Carré Spillway (BCS) opened twice in one year during 2019. The coastal cyanoHAB coincided with the second BCS opening. The main objectives of this study were: (1) to investigate the potential for using the National Aeronautics and Space Administration (NASA) ocean color standard Cyanobacteria Index (CIcyano) algorithm to characterize the spatial and temporal extent of the 2019 cyanoHAB; (2) to couple the CIcyano data with river discharge, salinity, and modeled-wind data to study the conditions leading to the cyanoHAB and factors aiding the advection and persistence of the bloom within the Mississippi Sound, including a possible relationship to the BCS; (3) to further investigate the relationship with the BCS by repeating the methods using data from 2018, which was a year when the BCS was opened but no evidence of cyanoHABs was reported along the Mississippi coast. Weekly means and monthly frequency CIcyano images, river discharge, salinity, and modeled-wind data from February to September of 2018 and 2019 were analyzed, which coincide with three BCS openings. In March 2018, a cyanobacteria bloom was observed within Lake Pontchartrain coinciding with the BCS opening; however, the month-long bloom was contained to the lake. Two distinct cyanoHABs were observed in 2019 and both blooms were advected into the Mississippi Sound, and likely contributed to the 3-month-long beach water advisories of 2019 along the Mississippi coastline. From March to mid-July 2019, salinity at stations within the Mississippi Sound was consistently near zero indicating high levels of freshwater. During that time, winds were predominantly northwestward, preventing the BCS waters from flushing into the Mississippi Shelf and resulting in BCS waters remaining longer within the estuarine lakes and Mississippi Sound. Although the BCS had an undeniable impact on the presence of the coastal cyanoHAB of 2019, other variables including wind direction, water flow, mixing, and persistence of freshwater within the Sound can determine the intensity and extent of the cyanoHABs. Coupling in situ phytoplankton information from freshwater water bodies to the marine continuum along with water flow, wind data, and satellite imagery could help identify cyanoHABs at early stages and forecast their trajectory and potential impacts on coastal areas.

Bonnet Carré Spillway↗
DOE OSTI2025

Green genes from blue greens: challenges and solutions to unlocking the potential of cyanobacteria in drug discovery

Cyanobacteria are prolific producers of biologically active compounds that are important in influencing ecology, behavior of interacting organisms, and as leads in drug discovery efforts. Here we discuss the challenges faced by all natural product researchers, especially those that focus on cyanobacteria, and then describe progress that has been made in these areas. We also propose some solutions, paths forward, and thoughts for consideration on these challenges.

Philmus, Benjamin↗
DOE OSTI2025

High-density CRISPRi screens reveal diverse routes to improved acclimation in cyanobacteria

Cyanobacteria are the oldest form of photosynthetic life on Earth and contribute to primary production in nearly every habitat, from permafrost to hot springs. Despite longstanding interest in the acclimation of these microbes, it remains poorly understood and challenging to rewire. Here, this study uses a high-density, genome-wide CRISPR interference screen to examine the influence of gene-specific transcriptional variation on the growth of Synechococcus sp. PCC 7002 under environmental extremes. Surprisingly, many partial knockdowns enhanced fitness under cold monochromatic conditions. Transcriptional repression of genes for core subunits of the NDH-1 complex, which are important for photosynthesis and carbon uptake, improved growth rates under both red and blue light but at distinct, color-specific optima. Most genes with fitness-improving knockdowns were distinct to each light color, and dual-target transcriptional repression produced nonadditive effects. Findings reveal diverse routes to improved acclimation in cyanobacteria (e.g., attenuation of genes involved in CO 2 uptake, light harvesting, translation, and purine metabolism) and provide an approach for using gradients in sgRNA activity to pinpoint biochemically influential transcriptional changes in cells.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Biocrust Mosses and Cyanobacteria Exhibit Distinct Carbon Uptake Responses to Variations in Precipitation Amount and Frequency

ABSTRACT Dryland organisms exhibit varied responses to changes in precipitation, including event size, frequency, and soil moisture duration, influencing carbon uptake and reserve management strategies. This principle, central to the pulse‐reserve paradigm, has not been thoroughly evaluated in biological soil crusts (biocrusts), essential primary producers on dryland surfaces. We conducted two experiments to investigate carbon uptake in biocrusts under different precipitation regimes. In the first, we applied a gradient of watering amounts to biocrusts dominated by moss or cyanobacteria, hypothesising distinct pulse‐response strategies. The second experiment extended watering treatments over three months, varying pulse size and frequency. Our results revealed distinct carbon uptake patterns: moss crusts exhibited increased CO 2 uptake with larger, less frequent watering events, whereas cyanobacteria crusts maintained similar carbon uptake across all event sizes. These findings suggest divergent pulse‐response strategies across biocrust types, with implications for modelling dryland carbon dynamics and informing land management under changing precipitation regimes.

Young, Kristina E. [USDA‐ARS Jornada Experimental ↗
NASA NTRS1985

The role of sulfur in osmoregulation and salinity tolerance in cyanobacteria, algae, and plants

Organosulfur compounds are involved in osmoregulation and salinity tolerance in some cyanobacteria and photosynthetic eukaryotes. Glycinebetaine, the osmolyte of the halotolerant cyanobacterium, Aphanothece halophytica, requires the sulfonium compound. S-adenosyl-methionine (SAM) for its synthesis. Glutamate is the nitrogen source, SAM is the methyl carbon and serine the carbon backbone source of this unique osmolyte. Inhibitor studies suggest that photorespiration interacts with sulfur metabolism to control betaine synthesis in cyanobacteria. The limiting factor for SAM synthesis is formate from photorespiration. SAM is, in turn, the methyl donor for betaine synthesis from serine. The nitrogen component of serine is from glutamate. Betaine synthesis is hypothesized to be regulated via potassium. The biosynthesis of dimethyl-B-propiothetin (DMPT, which is the same as beta-dimethyl sulfonioprpionate) and diacylsulfoquinovosylglycerol were elucidated as having their roles in osmoregulation and salinity tolerance. The relation between these sulfolipids and the sulfur cycle was discussed.

Yopp, J. H.↗
NASA NTRS1987

Origin and evolution of osmoregulatory mechanisms in blue-green algae (cyanobacteria) as a function of metabolic and structural complexity: Reflections of Precambrian paleobiology

Major accomplishments underlying the basic understanding of cyanobacterial resistance to salt tolerance and osmotic stress were made. The methodology proposed included: the tracing of the pathways of formation of osmoregulatory solutes by traditional methods involving C-14 labelled substrates; gas chromatography; amino acid analysis; X-ray analysis using scanning transmission electron microscopy; and most importantly, C-13 labelled substrates, followed by Nuclear Magnetic Resonance (NMR) spectroscopy. It was found that the cyanobacteria employ a diversity of organic, osmoregulatory solutes. Osmoregulatory solutes were found to serve four functions: adjustment of water activity, noninhibition of enzymes; lowering of K sub m of enzymes to allow functioning at normal levels when the intracellular salt accumulates, and extending the pH optimum of enzymes as intracellular pH rises due to proton-potassium ion pump action during osmoregulation. Differences in osmoregulatory solutes may, but are not always, be attributed to differences in nutritional capabilities. The mechanism of osmoregulation and concomitant salt tolerance in halophilic cyanobacteria was elucidated. The activities of betaine and S-Adenosylhomocysteine hydrolase are discussed.

Yopp, John H.↗
NASA NTRS2000

The Evolution of Sulfide Tolerance in the Cyanobacteria

Understanding how the function of extant microorganisms has recorded both their evolutionary histories and their past interactions with the environment is a stated goal of astrobiology. We are taking a multidisciplinary approach to investigate the diversification of sulfide tolerance mechanisms in the cyanobacteria, which vary both in their degree of exposure to sulfide and in their capacity to tolerate this inhibitor of photosynthetic electron transport. Since conditions were very reducing during the first part of Earth's history and detrital sulfides have been found in Archean sediments, mechanisms conferring sulfide tolerance may have been important for the evolutionary success of the ancestors of extant cyanobacteria. Two tolerance mechanisms have been identified in this group: (1) resistance of photosystem II, the principal target of sulfide toxicity; and (2) maintenance of the ability to fix carbon despite photosystem II inhibition by utilizing sulfide as an electron donor in photosystem I - dependent, anoxygenic photosynthesis. We are presently collecting comparative data on aspects of sulfide physiology for laboratory clones isolated from a variety of habitats. These data will be analyzed within a phylogenetic framework inferred from molecular sequence data collected for these clones to test how frequently different mechanisms of tolerance have evolved and which tolerance mechanism evolved first. In addition, by analyzing these physiological data together with environmental sulfide data collected from our research sites using microelectrodes, we can also test whether the breadth of an organism's sulfide tolerance can be predicted from the magnitude of variation in environmental sulfide concentration it has experienced in its recent evolutionary past and whether greater average sulfide concentration and/or temporal variability in sulfide favors the evolution of a particular mechanism of sulfide tolerance.

Miller, Scott R.↗
NASA NTRS1994

Distribution of cultivated and uncultivated cyanobacteria and Chloroflexus-like bacteria in hot spring microbial mats

Oligodeoxynucleotide hybridization probes were developed to complement specific regions of the small subunit (SSU) rRNA sequences of cultivated and uncultivated cyanobacteria and Chloroflexus-like bacteria, which inhabit hot spring microbial mats. The probes were used to investigate the natural distribution of SSU rRNAs from these species in mats of Yellowstone hot springs of different temperatures and pHs as well as changes in SSU rRNA distribution resulting from 1-week in situ shifts in temperature, pH, and light intensity. Synechococcus lividus Y-7c-s SSU rRNA was detected only in the mat of a slightly acid spring, from which it may have been initially isolated, or when samples from a more alkaline spring were incubated in the more acid spring. Chloroflexus aurantiacus Y-400-fl SSU rRNA was detected only in a high-temperature mat sample from the alkaline Octopus Spring or when lower-temperature samples from this mat were incubated at the high-temperature site. SSU rRNAs of uncultivated species were more widely distributed. Temperature distributions and responses to in situ temperature shifts suggested that some of the uncultivated cyanobacteria might be adapted to high-, moderate-, and low-temperature ranges whereas an uncultivated Chloroflexus-like bacterium appears to have broad temperature tolerance. SSU rRNAs of all uncultivated species inhabiting a 48 to 51 degrees C Octopus Spring mat site were most abundant in the upper 1 mm and were not detected below a 2.5-to 3.5-mm depth, a finding consistent with their possible phototrophic nature. However, the effects of light intensity reduction on these SSU rRNAs were variable, indicating the difficulty of demonstrating a phototrophic phenotype in light reduction experiments.

Non-NASA Center↗