Engineering Papers⌕ Search

SEARCH · Engineering Papers

Results for “gene flow”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 127 records · Page 7

New principles of self‐organization created through the interplay of DNA condensates, microtubules, and motors

Bioinspired design—which holds great promise for a new generation of materials that are robust to defects, scalable under green manufacture, environmentally responsive, and programmably reconfigurable—requires mastery over molecular self-organization. Yet, from its specific mechanisms to most general architectures, the principles governing self-organization remain poorly understood and not even fully enumerated. For living systems, one obvious architectural principle is the modular reuse of a few simple molecular components in myriad combinations to achieve more complex phenomena. For example, the mechanical tasks of a cell are driven by the nonequilibrium dynamics of cytoskeletal filaments and molecular motors—the same filaments and motors, reprogrammed by a variety of modulators, perform tasks ranging from cell movement to division. Similarly, many compartmentalization tasks are performed by liquid-like condensates of simple components, which act as membraneless organelles to localize particular molecules in space and time (e.g. for gene regulation or RNA processing). In a few cases, condensates combine and interact with the cytoskeleton to create still more complex phenomena, e.g. the nucleation of microtubule asters from the centrosome (a protein condensate) to form the mitotic spindle during cell division. Very little is known about the fundamental mechanisms of such filament-plus-condensate phenomena. Despite few examples, the landscape of behaviors that can be achieved through the combination of condensates, filaments, and motors appears vast. However, exploration has been hindered by a lack of systems that have sufficiently programmable and dynamically tunable interactions between component condensates, filaments, and motors. We proposed to combine programmable DNA condensates, filamentous microtubules, and light-controlled motors into self-organizing systems whose principles go beyond those that have been observed in nature. In one limit, our systems will use microtubules and motors to create the molecular analog of a network of roads, which will organize droplets of DNA condensates capable of carrying molecular cargo. DNA condensates coupled to motors will flow from one microtubule aster hub to another, with their direction and timing controlled by DNA circuits. In another limit, microtubules will swim through bulk DNA condensates and exhibit strong interactions with boundaries between different types of condensates. Microtubule swimmers will reflect, get trapped, or refract at boundaries, under a mechanical analog of the classical optical index of refraction. DNA condensates having different mechanical indexes of refraction will be used to construct the analog of optical lenses, so that microtubule swimmers can be manipulated like light—collimated, diffracted, focused, and sorted based on properties analogous to wavelength. These two limits define two new architectures, within which multiple new mechanistic principles for self-organization will be discovered and explored. To explore these architectures, the motor-based coupling between DNA condensates and filaments will be controlled in time and space through the use of opto-proteins that create reversible links between DNA condensates and motors upon illumination. For each principle of interest, patterns of light will create virtual experiments by defining patterns of activity where DNA condensates walk along filaments, or filaments swim through condensates, and patterns of inactivity which will serve either as controls, or as boundary conditions vital to create the desired phenomena. This research serves the goals of Basic Energy Sciences Biomolecular Material Program by elucidating the principles by which the emergent, nonequilibrium behavior of collections of DNA condensates, motors, and microtubules can be programmed by environmental light patterns to create complex motion and materials transport. Because DNA condensates can be readily coupled to virtually any high performance nanomaterial, from carbon nanotubes, to metal nanoparticles, to light harvesting systems, this work provides a path to the construction, self-maintenance and reconfiguration of materials relevant to the Department of Energy.

60 APPLIED LIFE SCIENCES↗

Nitric Oxide Formation by Meteoroids in the Upper Atmosphere

The process of nitric oxide formation during atmospheric entry of meteoroids is analyzed theoretically. An ablating meteoroid is assumed to be a point source in a uniform flow with a continuum regime evolving in its wake. The amount of nitric oxide produced by high-temperature reactions of air in the continuum regime is calculated by numerical integration of chemical-rate equations. This is accomplished by assuming that flow properties are constant across the reacting region, the radius of the region being determined from considerations of shock-wave formation and molecular diffusion. The results, when summed over the observed mass, velocity, and entry-angle distributions of meteoroids, provide annual global production rates of nitric oxide as a function of altitude. The peak production of nitric oxide is found to occur at altitudes between 9 x 10(exp 4) and 10(exp 5) m, the total annual rate being about 4 x 10(exp 7) kg. The present results suggest that the large concentration of nitric oxide observed below 9.5 x 10(exp 4) m could be attributed to meteoroids instead of photodissociation of nitrogen into metastable, 2D-state atoms, as has been previously hypothesized.

Menees, Gene P.↗

Enhanced Resistance Pines for Improved Renewable Biofuel and Chemical Production (Technical Report)

We completed phenotyping constitutive and inducible oleoresin flow across two seasons, constitutive resin canal number and density and wood terpene content in our ADEPT2 and CCLONES populations. We completed genetic association between 19 oleoresin phenotypes and a total of 523,192 SNP markers from ADEPT2 and 13,883 SNP markers in CCLONES using four mixed linear models. A total of 293 significant SNPs (FDR = 0.20) were identified. We used the MENTOR tool to mine mechanistic connections from a multiplex network constructed from poplar multi-omic data to construct a conceptual model for a subset of these significant SNPs. Our model contains 6 transcriptional regulators in addition to 3 monoterpene synthases. To generate more lines of evidence for these significant SNPs, we completed a time course RNAseq experiment after inducing vascular zone cells to differentiate into new resin canals with a methyl jasmonate treatment, a single nuclei RNAseq that identified differentiating resin canal epithelial cells and are completing analysis for a QTL study in a hybrid pine population. The time course identified 4634 significantly down and 1890 significantly up regulated transcripts after treatment with methyl jasmonate, an inducer of new resin canal formation in the vascular cambial meristem. To analyze this large set of differentially regulated genes, we created a predictive expression network and analyzed it with random walk restart using 6 seed genes coding for transcription factors regulating xylem differentiation in poplar. Of the top ranked 200 transcripts, 119 transcripts were significant differentially expressed supporting these transcripts as potential candidates regulating resin canal formation. Analysis of single nuclei sequencing of shoot tips that contain differentiating resin canals, identified 10 clusters. One cluster was highly enriched in transcripts coding for 9 of the enzymes in the MEP pathway 3 prenyl synthetases, and 3 monoterpene synthases strongly suggesting that this cluster represents resin canal epithelial cells. We are mining the additional transcripts to create a trajectory analysis. In summary, we have identified > 10 novel genes that are strongly supported candidates for further analysis in breeding lines and for genetic engineering over- and under- expressing lines to increase wood terpene content to improve resistance to insect and fungal pathogens while simultaneously increasing terpene supplies for renewable chemicals and biofuels.

59 BASIC BIOLOGICAL SCIENCES↗

Wave combustors for trans-atmospheric vehicles

The Wave Combustor is an airbreathing hypersonic propulsion system which utilizes shock and detonation waves to enhance fuel-air mixing and combustion in supersonic flow. In this concept, an oblique shock wave in the combustor can act as a flameholder by increasing the pressure and temperature of the air-fuel mixture and thereby decreasing the ignition delay. If the oblique shock is sufficiently strong, then the combustion front and the shock wave can couple into a detonation wave. In this case, combustion occurs almost instantaneously in a thin zone behind the wave front. The result is a shorter, lighter engine compared to the scramjet. This engine, which is called the Oblique Detonation Wave Engine (ODWE), can then be utilized to provide a smaller, lighter vehicle or to provide a higher payload capability for a given vehicle weight. An analysis of the performance of a conceptual trans-atmospheric vehicle powered by an ODWE is given here.

Menees, Gene P.↗

Integrated CMOS photodetectors and signal processing for very low-level chemical sensing with the bioluminescent bioreporter integrated circuit

We report an integrated CMOS microluminometer optimized for the detection of low-level bioluminescence as part of the bioluminescent bioreporter integrated circuit (BBIC). This microluminometer improves on previous devices through careful management of the sub-femtoampere currents, both signal and leakage, that flow in the front-end processing circuitry. In particular, the photodiode is operated with a reverse bias of only a few mV, requiring special attention to the reset circuitry of the current-to-frequency converter (CFC) that forms the front-end circuit. We report a sub-femtoampere leakage current and a minimum detectable signal (MDS) of 0.15 fA (1510 s integration time) using a room temperature 1.47 mm2 CMOS photodiode. This microluminometer can detect luminescence from as few as 5000 fully induced Pseudomonas fluorescens 5RL bacterial cells. c2002 Elsevier Science B.V. All rights reserved.

Non-NASA Center↗

Irradiated HMEC from A-T Heterozygous Breast Tissue

Women who are heterozygous for ataxia-telangiectasia (A-T) carry a single defective ATM gene in chromosome 11 q22-23, and have been statistically determined with high significance within a defined database to be approximately 5-fold more susceptible for developing breast cancer than their noma1 counterpart. Breast cancer susceptibility of these A-T heterozygotes has been hypothesized to include consequence of response to damage caused by low levels of ionizing radiation. Prophylactic mastectomy specimens were donated by a 41 year-old obligate A-T heterozygote who was located prior to her elective surgery through an existing pedigree. Harvest of that breast tissue provided an isolate of long-term growth human mammary epithelial cells (HMEC), designated WH612/3. An isolate of presumed normal long-term growth HMEC, designated 48R, was obtained from Dr. Martha Stampfer (Lawrence Berkeley Laboratory, University of California), and the A-T heterozygous HMEC were transformed with E6 and E7 oncogenes of human papilloma virus Type-16 in the laboratory of Dr. Ray White (Hunt- Cancer Institute, University of Utah) for use in this study. The objective of this study is to study the expression of end points that may bear on cancer outcome following irradiation of HMEC. Specific end points are cell survival, cell cycle, p53 expression, and apoptosis. Survival curves, immunostaining, and flow cytometery are used to examine these end points. Radiation-induced cell killing shows less shoulder development in the survival curve for WH61U3 compared to 48R HMEC, suggesting less repair of damage in the former HMEC. Additional information is included in the original extended abstract.

Richmond, Robert↗

Experimental and analytical studies in fluids

The first objective was to analyze and design a true airspeed sensor which will replace the conventional pitot-static pressure transducer for small commercial aircraft. The second objective was to obtain a numerical solution and predict the frequency response which is generated by the vortex whistle at a certain airspeed. It was concluded flow rate measurements indicate that the vortex tube sound frequency is linearly proportional to the frequency response. The vortex tube whistle frequency is dependent upon geometrical parameters to such an extent that: an increase in vortex tube length produces a decrease in frequency response and that an increase in the exhaust nozzle length produces an increase in the frequency precession. An increase in the vortex tube diameter produces a decrease in frequency precession. An increase in swirler diameter produces a decrease in frequency. An increase in the location distance of the microphone pickup signal point from the inside edge of the exit nozzle produces an increase in frequency response. The experimental results indicate that those parameters most significantly effecting frequency are in descending order of importance microphone location, vortex tube diameter, exit nozzle length, vortex tube length, and swirler diameter.

Goglia, Gene L.↗

Acoustic and Laser Doppler Anemometer Results for Confluent, 22-Lobed, and Unique-Lobed Mixer Exhaust Systems for Subsonic Jet Noise Reduction

The objective of this task was to develop a design methodology and noise reduction concepts for high bypass exhaust systems which could be applied to both existing production and new advanced engine designs. Special emphasis was given to engine cycles with bypass ratios in the range of 4:1 to 7:1, where jet mixing noise was a primary noise source at full power takeoff conditions. The goal of this effort was to develop the design methodology for mixed-flow exhaust systems and other novel noise reduction concepts that would yield 3 EPNdB noise reduction relative to 1992 baseline technology. Two multi-lobed mixers, a 22-lobed axisymmetric and a 21-lobed with a unique lobe, were designed. These mixers along with a confluent mixer were tested with several fan nozzles of different lengths with and without acoustic treatment in GEAE's Cell 41 under the current subtask (Subtask C). In addition to the acoustic and LDA tests for the model mixer exhaust systems, a semi-empirical noise prediction method for mixer exhaust system is developed. Effort was also made to implement flowfield data for noise prediction by utilizing MGB code. In general, this study established an aero and acoustic diagnostic database to calibrate and refine current aero and acoustic prediction tools.

Salikuddin, M.↗

[The role of the serotonin system in the stress response of various cells]

The recombinant mouse brain serotonin receptor (5HT1c) was used to study the response of plant cells and oocytes to a stress signal activated by the serotonin-serotonin receptor interaction and associated Ca2+ flow. Based on plant expression vectors, recombinant constructs were obtained to direct production of 5HT1c fused with the green fluorescent protein in plant cells. The mRNAs for hybrid proteins were synthesized in an in vitro transcription system. The expression and function of the hybrid protein and the function of the associated ion channels were electrophysiologically studied in Xenopus laevis oocytes injected with the hybrid mRNA. The hybrid protein was functional and changed the operation of the Ca2+ channel in oocytes. To study the expression of the hybrid constructs in plant cells, the in vitro transcription product was inoculated in tobacco leaves, which then fluoresced.

Non-NASA Center↗

Insights into the Processing of Carbon by Early Microbial Ecosystems

Interactions between Earth and the biosphere that were crucial for early biological evolution also influenced substantially the processes that circulate C between its reservoirs in the atmosphere, ocean, crust and mantle. The C-13 C-12 values of crustal carbonates and organics have recorded changes both in biological discrimination and in the relative rates of burial of organics and carbonates. A full interpretation of these patterns needs further isotopic studies of microbial ecosystems and individual anaerobes. Thus we measured carbon isotope discrimination during autotrophic and heterotrophic growth of pure cultures of sulfate-reducing bacteria and archaea (SRB and SRA). Discrimination during CO2 assimilation is significantly larger than during heterotrophic growth on lactate or acetate. SRB grown lithoautotrophically consumed less than 3% of available CO2 and exhibited substantial discrimination, as follows: Desulfobacterium autotrophicum (alpha 1.0100 to 1.0123), Desulfobacter hydrogenophilus (alpha = 0.0138), and Desulfotomuculum acetoxidans (alpha = 1.0310). Mixotrophic growth of Desulfovibrio desulfuricans on acetate and CO2 resulted in biomass with delta C-13 composition intermediate to that of the substrates. We have recently extended these experiments to include the thermophilic SRA Archeoglobus spp. Ecological forces also influence isotopic discrimination. Accordingly, we quantified the flow of C and other constituents in modern marine cyanobacterial mats, whose ancestry extends back billions of years. Such ecosystem processes shaped the biosignatures that entered sediments and atmospheres. At Guerrero Negro, BCS, Mexico, we examined mats dominated by Microcoleus (subtidal) and Lyngbya (intertidal to supratidal) cyanobacteria. During 24 hour cycles, we observed the exchange of O2 and dissolved inorganic C (DIC) between mats and the overlying water. Microcoleus mats assimilated near-equal amounts of DIC during the day as they released at night, but Lyngbya mats typically showed net uptake of DIC over the diel cycle. Patterns of O2 daytime release and nighttime uptake mirrored these DIC trends in both mat types. Nighttime DIC effluxes from Microcoleus mats were equivalent in the presence versus absence of O2, whereas nighttime DIC effluxes from Lyngbya mats dropped markedly in the absence of O2. Thus aerobic diagenesis was more important in Lyngbya mats than in Microcoleus mats, perhaps because trapped O2 bubbles persist only in Lyngbya mats at night and thus sustain populations of aerbes. In both mat types, effluxes of H2, CH4 and short-chain fatty acids were much greater at night in the absence of 02, emphasizing the importance of fermentation. Differences observed between Microcoleus versus Lyngbya mats forecast differences in their microbial populations and in their patterns of gene expression.

DesMarais, D.↗

Characterization of Microbial Communities Found in Bioreactor Effluent

The purpose of this investigation was to examine microbial communities of simulated wastewater effluent from hollow fiber membrane bioreactors collected from the Space Life Science Laboratory and Texas Technical University. Microbes were characterized using quantitative polymerase chain reaction where a total count of bacteria and fungi were determined. The primers that were used to determine the total count of bacteria and fungi were targeted for 16S rDNA genes and the internal transcribed spacer, respectively. PCR products were detected with SYBR Green I fluorescent dye and a melting curve analysis was performed to identify unique melt profiles resulting from DNA sequence variations from each species of the community. Results from both the total bacteria and total fungi count assays showed that distinct populations were present in isolates from these bioreactors. This was exhibited by variation in the number of peaks observed on the melting curve analysis graph. Further analysis of these results using species-specific primers will shed light on exactly which microbes are present in these effluents. Information gained from this study will enable the design of a system that can efficiently monitor microbes that play a role in the biogeochemical cycling of nitrogen in wastewater on the International Space Station to assist in the design of a sustainable system capable of converting this nutrient.

Flowe, Candice↗

Wave combustors for trans-atmospheric vehicles

A performance analysis is given of a conceptual transatmospheric vehicle (TAV). The TAV is powered by a an oblique detonation wave engine (ODWE). The ODWE is an airbreathing hypersonic propulsion system which utilizes shock and detonation waves to enhance fuel-air mixing and combustion in supersonic flow. In this wave combustor concept, an oblique shock wave in the combustor can act as a flameholder by increasing the pressure and temperature of the air-fuel mixture, thereby decreasing the ignition delay. If the oblique shock is sufficiently strong, then the combustion front and the shock wave can couple into a detonation wave. In this case, combustion occurs almost instantaneously in a thin zone behind the wave front. The result is a shorter lighter engine compared to the scramjet. The ODWE-powered hypersonic vehicle performance is compared to that of a scramjet-powered vehicle. Among the results outlined, it is found that the ODWE trades a better engine performance above Mach 15 for a lower performance below Mach 15. The overall higher performance of the ODWE results in a 51,000-lb weight savings and a higher payload weight fraction of approximately 12 percent.

Menees, Gene P.↗

Identification of Ser-543 as the major regulatory phosphorylation site in spinach leaf nitrate reductase

Spinach leaf NADH:nitrate reductase (NR) responds to light/dark signals and photosynthetic activity in part as a result of rapid regulation by reversible protein phosphorylation. We have identified the major regulatory phosphorylation site as Ser-543, which is located in the hinge 1 region connecting the cytochrome b domain with the molybdenum-pterin cofactor binding domain of NR, using recombinant NR fragments containing or lacking the phosphorylation site sequence. Studies with NR partial reactions indicated that the block in electron flow caused by phosphorylation also could be localized to the hinge 1 region. A synthetic peptide (NR6) based on the phosphorylation site sequence was phosphorylated readily by NR kinase (NRk) in vitro. NR6 kinase activity tracked the ATP-dependent inactivation of NR during several chromatographic steps and completely inhibited inactivation/phosphorylation of native NR in vitro. Two forms of NRk were resolved by using anion exchange chromatography. Studies with synthetic peptide analogs indicated that both forms of NRk had similar specificity determinants, requiring a basic residue at P-3 (i.e., three amino acids N-terminal to the phosphorylated serine) and a hydrophobic residue at P-5. Both forms are strictly calcium dependent but belong to distinct families of protein kinases because they are distinct immunochemically.

NASA Discipline Plant Biology↗

Open Science for Life in Space: Data Sharing and Tools for Knowledge Discovery

The next era in human space exploration is rapidly approaching and will require the use of countermeasures to deep space health hazards. The development of countermeasures (or, there-purposing of existing agents) will be highly dependent on our understanding of basic biological responses to space stressors (e.g. ionizing radiation, altered gravitational fields, altered day-night cycles, confinement, isolation, hostile-closed environments, distance-duration from Earth, exposure to celestial regolith, etc.). The fast-growing array of space biological data, which in the past was simply archived after minimal analysis, holds great potential if it can be reorganized and formatted for Open Science. Organizing the data for such analysis is a challenge because of its diverse nature (molecular, cellular, tissue, imaging, whole organism and behavior). We will discuss here several strategies that NASA's Biological and Physical Science Division has put in place to maximize the return on investment for spaceflight bioscience data. Open Science, as a scientific philosophy, is the concept that the more people who have access to the data, the more knowledge will be gained from it. This guiding principle led NASA to develop GeneLab in 2015. GeneLab houses spaceflight and relevant ground-based multi-omics data, and has grown to ~400 transcriptomatic, proteomic, metabolomic and epigenomic datasets from plant, rodent, small animal, and microbial space experiments. GeneLab provides users with various tools for data analysis and a visualization portal that allows users to interact with gene expression data from space-related 'omics experiments. Open Science is also about building scientific communities, and with this spirit in mind, GeneLab has spawned several Analysis Working Groups (AWGs), comprised of more than 200 volunteer scientists. The AWGs initially provided feedback on the processing pipeline and metadata 'omics standards for GeneLab. Over the last few years, they have become a community-driven science enterprise, engaging in large meta-analysis of GeneLab datasets, resulting in 10 publications (beyond the originally submitted research). Overall, the Open Science nature of GeneLab has resulted in a high degree of data-use, resulting in 40 enabled publications by open data. The enormous success and knowledge gained from GeneLab has led to a collection of sister NASA "Open Science Data Repositories (OSDR)" and research support groups. These include the NASA Ames Life Sciences Data Archive (ALSDA), the NASA Biological Institutional Scientific Collection (NBISC), and the Biospecimen Sharing Program (BSP). All are adopting the GeneLab data architecture system to maximize open-access, find-ability, accessibility, interoperability, and reusability (FAIR). ALSDA collects and curates phenotypic-physiological bioimaging-behavioral data from space and space-relevant non-human experiments, oftentimes coming from the same omics-associated experimental datasets found in GeneLab. Since 2021, a community of ~100 researchers have rallied around ALSDA, to provide feedback in a new ALSDA AWG focused on phenotypic-physiological investigation-sample-assay metadata standards (e.g., Micro-Computed Tomography, Light/Flourescence Microscopy, Western Blot, Flow Cytometry, Novel Object Recognition, Elevated Plus Maze, etc. of ~50 assays collected). These standards are part of a new single point-of-entry data submission portal for all non-human Space Biology and Human Research Program principal investigators, to submit, curate, and share their research data. With open-access space biological data now collected and curated together with rich metadata, and with the potential for linkage to "big data" from the international biological and medical communities (NIH, EBI, etc.), the artificial intelligence and machine learning (AI/ML) era has started for Space Biology.

omics↗

Open Science for Life in Space: Data Sharing and Tools for Knowledge Discovery

The next era in human space exploration is rapidly approaching and will require the use of countermeasures to deep space health hazards. The development of countermeasures (or, the re-purposing of existing agents) will be highly dependent on our understanding of basic biological responses to space stressors (e.g. ionizing radiation, altered gravitational fields, altered day-night cycles, confinement, isolation, hostile-closed environments, distance-duration from Earth, exposure to celestial regolith, etc.). The fast-growing array of space biological data, which in the past was simply archived after minimal analysis, holds great potential if it can be reorganized and formatted for Open Science. Organizing the data for such analysis is a challenge because of its diverse nature (molecular, cellular, tissue, imaging, whole organism and behavior). We will discuss here several strategies that NASA’s Biological and Physical Science Division has put in place to maximize the return on investment for spaceflight bioscience data. Open Science, as a scientific philosophy, is the concept that the more people who have access to the data, the more knowledge will be gained from it. This guiding principle led NASA to develop GeneLab in 2015. GeneLab houses spaceflight and relevant ground-based multi-omics data, and has grown to ~400 transcriptomic, proteomic, metabolomic and epigenomic datasets from plant, rodent, small animal, and microbial space experiments. GeneLab provides users with various tools for data analysis and a visualization portal that allows users to interact with gene expression data from space-related ‘omics experiments. Open Science is also about building scientific communities, and with this spirit in mind, GeneLab has spawned several Analysis Working Groups (AWGs), comprised of more than 200 volunteer scientists. The AWGs initially provided feedback on the processing pipeline and metadata ‘omics standards for GeneLab. Over the last few years, they have become a community-driven science enterprise, engaging in large meta-analysis of GeneLab datasets, resulting in 10 publications (beyond the originally submitted research). Overall, the Open Science nature of GeneLab has resulted in a high degree of data re-use, resulting in 38 additional publications derived from the original 67 publication over the past four years. The enormous success and knowledge gained from GeneLab has led to a collection of sister NASA “Open Science Data Repositories (OSDR)” and research support groups. These include the NASA Ames Life Sciences Data Archive (ALSDA), the NASA Biological Institutional Scientific Collection (NBISC), and the Biospecimen Sharing Program (BSP). All are adopting the GeneLab data architecture system to maximize open-access, find-ability, accessibility, interoperability, and reusability (FAIR). ALSDA collects and curates phenotypic-physiological bioimaging-behavioral data from space and space-relevant non-human experiments, oftentimes coming from the same omics-associated experimental datasets found in GeneLab. Since 2021, a community of ~100 researchers have rallied around ALSDA, to provide feedback in a new ALSDA AWG focused on phenotypic-physiological investigation-sample-assay metadata standards (e.g., Micro-Computed Tomography, Light/Fluorescence Microscopy, Western Blot, Flow Cytometry, Novel Object Recognition, Elevated Plus Maze, etc. of ~50 assays collected). These standards are part of a new single point-of-entry data submission portal for all non-human Space Biology and Human Research Program principal investigators, to submit, curate, and share their research data. With open-access space biological data now collected and curated together with rich metadata, and with the potential for linkage to “big data” from the international biological and medical communities (NIH, EBI, etc.), the artificial intelligence and machine learning (AI/ML) era has started for Space Biology. Several other talks will cover these topics in this conference.

life sciences↗

Selected contribution: a three-dimensional model for assessment of in vitro toxicity in balaena mysticetus renal tissue

This study established two- and three-dimensional renal proximal tubular cell cultures of the endangered species bowhead whale (Balaena mysticetus), developed SV40-transfected cultures, and cloned the 61-amino acid open reading frame for the metallothionein protein, the primary binding site for heavy metal contamination in mammals. Microgravity research, modulations in mechanical culture conditions (modeled microgravity), and shear stress have spawned innovative approaches to understanding the dynamics of cellular interactions, gene expression, and differentiation in several cellular systems. These investigations have led to the creation of ex vivo tissue models capable of serving as physiological research analogs for three-dimensional cellular interactions. These models are enabling studies in immune function, tissue modeling for basic research, and neoplasia. Three-dimensional cellular models emulate aspects of in vivo cellular architecture and physiology and may facilitate environmental toxicological studies aimed at elucidating biological functions and responses at the cellular level. Marine mammals occupy a significant ecological niche (72% of the Earth's surface is water) in terms of the potential for information on bioaccumulation and transport of terrestrial and marine environmental toxins in high-order vertebrates. Few ex vivo models of marine mammal physiology exist in vitro to accomplish the aforementioned studies. Techniques developed in this investigation, based on previous tissue modeling successes, may serve to facilitate similar research in other marine mammals.

Whales/genetics/physiology↗

Biosensors for the detection of chorismate and cis,cis -muconic acid in Corynebacterium glutamicum

Abstract Corynebacterium glutamicum ATCC 13032 is a promising microbial chassis for industrial production of valuable compounds, including aromatic amino acids derived from the shikimate pathway. In this work, we developed two whole-cell, transcription factor based fluorescent biosensors to track cis,cis-muconic acid (ccMA) and chorismate in C. glutamicum. Chorismate is a key intermediate in the shikimate pathway from which value-added chemicals can be produced, and a shunt from the shikimate pathway can divert carbon to ccMA, a high value chemical. We transferred a ccMA-inducible transcription factor, CatM, from Acinetobacter baylyi ADP1 into C. glutamicum and screened a promoter library to isolate variants with high sensitivity and dynamic range to ccMA by providing benzoate, which is converted to ccMA intracellularly. The biosensor also detected exogenously supplied ccMA, suggesting the presence of a putative ccMA transporter in C. glutamicum, though the external ccMA concentration threshold to elicit a response was 100-fold higher than the concentration of benzoate required to do so through intracellular ccMA production. We then developed a chorismate biosensor, in which a chorismate inducible promoter regulated by natively expressed QsuR was optimized to exhibit a dose-dependent response to exogenously supplemented quinate (a chorismate precursor). A chorismate–pyruvate lyase encoding gene, ubiC, was introduced into C. glutamicum to lower the intracellular chorismate pool, which resulted in loss of dose dependence to quinate. Further, a knockout strain that blocked the conversion of quinate to chorismate also resulted in absence of dose dependence to quinate, validating that the chorismate biosensor is specific to intracellular chorismate pool. The ccMA and chorismate biosensors were dually inserted into C. glutamicum to simultaneously detect intracellularly produced chorismate and ccMA. Biosensors, such as those developed in this study, can be applied in C. glutamicum for multiplex sensing to expedite pathway design and optimization through metabolic engineering in this promising chassis organism. One-Sentence Summary High-throughput screening of promoter libraries in Corynebacterium glutamicum to establish transcription factor based biosensors for key metabolic intermediates in shikimate and β-ketoadipate pathways.

59 BASIC BIOLOGICAL SCIENCES↗

Proliferation, differentiation and apoptosis in connexin43-null osteoblasts

Osteoblasts are highly coupled by gap junctions formed primarily by connexin43 (Cx43). We have shown that interference with Cx43 expression or function disrupts transcriptional regulation of osteoblast genes, and that deletion of Cx43 in the mouse causes skeletal malformations, delayed mineralization, and osteoblast dysfunction. Here, we studied the mechanisms by which genetic deficiency of Cx43 alters osteoblast development. While cell proliferation rates were similar in osteoblastic cells derived from calvaria of Cx43-null and wild type mice, camptothecin-induced apoptosis was 3-fold higher in mutant compared to wild type osteoblasts. When grown in mineralizing medium, Cx43-null cells were able to produce mineralized matrix but it took one week longer to reach the same mineralization levels as in normal cells. Likewise, expression of alkaline phosphatase activity per cell--a marker of osteoblast differentiation--was maximal only 2 weeks later in Cx43-null relative to wild-type cells. These observations suggest that Cx43 is important for a normal and timely development of the osteoblastic phenotype. Delayed differentiation and increase programmed cell death may explain the skeletal phenotype of Cx43-null mice.

NASA Discipline Cell Biology↗