Results for “sequencing test” — Page 52

NASA NTRS1992

Evidence of orbital forcing in 510 to 530 million year old shallow marine cycles, Utah and western Canada

Spectral analyses of two sequences of shallow marine sedimentary cycles that were deposited between 510 and 530 million years ago were completed. One sequence is from Middle Cambrian rocks in southern Utah and the other is from Upper Cambrian rocks in the southern Canadian Rockies. In spite of the antiquity of these strata, and even though there are differences in the age, location, and cycle facies between the two sequences, both records have distinct spectral peaks with surprisingly similar periodicities. A null model constructed to test for significance of the spectral peaks and circulatory in the methodology indicates that all but one of the spectral peaks are significant at the 90 percent confidence level. When the ratios between the statistically significant peaks are measured, we find a consistent relation to orbital forcing; specifically, the spectral peak ratios in both the Utah and Canadian examples imply that a significant amount of the variance in the cyclic records is driven by the short eccentricity (approximately 109 ky) and by the precessional (approximately 21 ky) components of the Earth's orbital variations. Neither section contains a significant component of variance at the period of the obliquity cycle, however.

Bond, Gerard C.↗

NASA NTRS2009

Ares I First Stage Booster Deceleration System: An Overview

In 2005, the Congressional NASA Authorization Act enacted a new space exploration program, the "Vision for Space Exploratien". The Constellation Program was formed to oversee the implementation of this new mission. With an intent not simply to support the International Space Station, but to build a permanent outpost on the Moon and then travel on to explore ever more distant terrains, the Constellation Program is supervising the development of a brand new fleet of launch vehicles, the Ares. The Ares lineup will include two new launch vehicles: the Ares I Crew Launch Vehicle and the Ares V Cargo Launch Vehicle. A crew exploration vehicle, Orion, will be launched on the Ares I. It will be capable of docking with the Space Station, the lunar lander, Altair, and the Earth Departure Stage of Ares V. The Ares V will be capable of lifting both large-scale hardware and the Altair into space. The Ares First Stage Team is tasked with developing the propulsion system necessary to liftoff from the Earth and loft the entire Ares vehicle stack toward low Earth orbit. The Ares I First Stage booster is a 12-foot diameter, five-segment, reusable solid rocket booster derived from the Space Shuttle's four segment reusable solid rocket booster (SRB). It is separated from the Upper Stage through the use of a Deceleration Subsystem (DSS). Booster Tumble Motors are used to induce the pitch tumble following separation from the Upper Stage. The spent Ares I booster must be recoverable using a parachute deceleration system similar to that of the Shuttle SRB heritage system. Since Ares I is much heavier and reenters the Earth's atmosphere from a higher altitude at a much higher velocity than the SRB, all of the parachutes must be redesigned to reliably meet the operational requisites of the new launch vehicles. This paper presents an overview of this new booster deceleration system. It includes comprehensive detail of the parachute deceleration system, its design and deployment sequences, including how and why it is being developed, the requirements it must meet, and the testing involved in its implementation.

King, Ron↗

NASA NTRS2016

Performance of an Automated System for Control of Traffic in Terminal Airspace

This paper examines the performance of a system that performs automated conflict resolution and arrival scheduling for aircraft in the terminal airspace around major airports. Such a system has the potential to perform separation assurance and arrival sequencing tasks that are currently handled manually by human controllers. The performance of the system is tested against several simulated traffic scenarios that are characterized by the rate at which air traffic is metered into the terminal airspace. For each traffic scenario, the levels of performance that are examined include: number of conflicts predicted to occur, types of resolution maneuver used to resolve predicted conflicts, and the amount of delay for all flights. The simulation results indicate that the percentage of arrivals that required a maneuver that changes the flight's horizontal route ranged between 11% and 15% in all traffic scenarios. That finding has certain implications if this automated system were to be implemented simply as a decision support tool. It is also found that arrival delay due to purely wake vortex separation requirements on final approach constituted only between 29% and 35% of total arrival delay, while the remaining major portion of it is mainly due to delay back propagation effects.

air traffic control↗

NASA NTRS2019

An Experimental Investigation concerning the Effects of AFP Defects on Progressive Failure of Tensile Coupons

This paper discusses characterization approaches for the evaluation of potential defects that may occur during the automated fiber placement (AFP) process. Two nondestructive evaluation techniques were utilized simultaneously, acoustic emission (AE) and digital image correlation (DIC), to investigate correlations between mechanical properties and damage propagation during tension tests. The layup configuration is designed with the defect running across the width, and the stacking sequence with seven layers is produced: [−45/0/45/̅9̅̅0̅]s. A total of 3 panels were manufactured: one containing a gap defect in the middle layer, another one with an overlap defect, and a pristine panel for baseline comparison. Each panel accommodates six coupons having dimensions of 20” x 2”. The results indicate that defects caused during the manufacturing process affect damage initiation and growth.

Anay, Rafal↗

NASA NTRSDate not supplied

ISS External Microorganisms: Collecting Planetary Protection Samples During Extravehicular Activity

Introduction: We have designed1, built, and tested a sampling kit (Fig. 1) to aseptically collect microbiological samples from exterior surfaces on the ISS (International Space Station). The kit was flown to ISS as part of the NG-19 commercial cargo mission in August of 2023. Astronauts will use the kit to collect samples from six exterior surfaces on ISS. These samples will be frozen at -80°C after collection and returned to the ground for next generation DNA sequencing. The results of this experiment will help inform planetary protection requirements for crewed missions. We hypothesize that there are detectable microbial communities outside ISS and that these communities originate from inside ISS. Current life support systems do not include any components for reducing microbial leakage. Gases are vented from inside ISS without filtration and there are currently no protocols in place to minimize bioburden on space suit exteriors prior to use. It is important to quantify the bioburden on current vehicles so that achievable limits can be set for crewed missions to astrobiologically relevant locations like Mars. Figure 1: The sampling kit contains eight swab canisters in total (six on the top and two on the bottom). A resuable end effector (1) is used to remove and reinstall the swabs. The kit also contains a handrail (2), tether loop (3), and, bayonet probes(4) to allow easy manipulation during EVA. Methods: We have designed a kit to carry 8 commercially available foam swabs into and out of vacuum without compromising the swabs’ sterility. The specially designed swab canisters contain a 0.2 µm Teflon filter that allows the canister to accommodate pressure changes without introducing unwanted contaminants. The kit meets existing EVA (Extravehicular Activity) safety requirements and has been used by human test subjects in the neutral buoyance lab and at vacuum in test chambers at the Johnson Space Center. In the early part of 2024, astronauts will use this sampling kit to swab six surfaces on the exterior of ISS. We will collect samples from: the airlock vestibule, the interior surface of the airlock thermal cover, an exterior handrail, a vent connected to the CO2 removal system, and a vent connected to the payload vacuum system inside ISS. Approximately 300 cm2 will be swabbed at each location. A seventh swab will be exposed to the vacuum of space without touching any surfaces as a blank. The eighth swab will remain sealed as a process control. After the EVA the swab kit will be frozen at -80°C and returned to earth at the earliest possible opportunity. The samples will remain frozen until they are thawed for next generation DNA sequencing on earth. We will use amplicon sequencing to identify any bacteria, archaea or fungi present in the samples. If there is enough DNA present, we will use shotgun metagenomic sequencing to further characterize the microbial ecology outside ISS. Preliminary Results: Results from ground-based testing demonstrate that the swab kit is capable of cycling in and out of vacuum without contaminating the swabs. Test subjects, wearing flight-like EVA gloves could remove swabs from the canister and sample discrete locations without inadvertently touching any other surfaces. Several types of bacteria and fungi were collected during these ground tests and survived up to 6 hours at vacuum. This includes non-spore forming bacterial like Staphyloccus capitis that are not traditionally considered extremophiles. Shotgun metagenomic sequencing of the ground-test samples revealed bacteria associated with human skin and airways on the exterior of space suits used during these tests 2 In addition to these results we will present preliminary results from our space-flight samples. We will also present lessons learned from attempting to collect microbiological samples during an EVA and describe how our results will affect planetary protection requirements for future crewed missions.

Aaron B. Regberg↗

NASA NTRSDate not supplied

ISS External Microorganisms: Collecting Planetary Protection Samples During Extravehicular Activity

Introduction: We have designed1, built, and tested a sampling kit (Fig. 1) to aseptically collect microbiological samples from exterior surfaces on the ISS (International Space Station). The kit was flown to ISS as part of the NG-19 commercial cargo mission in August of 2023. Astronauts will use the kit to collect samples from six exterior surfaces on ISS. These samples will be frozen at -80°C after collection and returned to the ground for next generation DNA sequencing. The results of this experiment will help inform planetary protection requirements for crewed missions. We hypothesize that there are detectable microbial communities outside ISS and that these communities originate from inside ISS. Current life support systems do not include any components for reducing microbial leakage. Gases are vented from inside ISS without filtration and there are currently no protocols in place to minimize bioburden on space suit exteriors prior to use. It is important to quantify the bioburden on current vehicles so that achievable limits can be set for crewed missions to astrobiologically relevant locations like Mars. Figure 1: The sampling kit contains eight swab canisters in total (six on the top and two on the bottom). A resuable end effector (1) is used to remove and reinstall the swabs. The kit also contains a handrail (2), tether loop (3), and, bayonet probes(4) to allow easy manipulation during EVA. Methods: We have designed a kit to carry 8 commercially available foam swabs into and out of vacuum without compromising the swabs’ sterility. The specially designed swab canisters contain a 0.2 µm Teflon filter that allows the canister to accommodate pressure changes without introducing unwanted contaminants. The kit meets existing EVA (Extravehicular Activity) safety requirements and has been used by human test subjects in the neutral buoyance lab and at vacuum in test chambers at the Johnson Space Center. In the early part of 2024, astronauts will use this sampling kit to swab six surfaces on the exterior of ISS. We will collect samples from: the airlock vestibule, the interior surface of the airlock thermal cover, an exterior handrail, a vent connected to the CO2 removal system, and a vent connected to the payload vacuum system inside ISS. Approximately 300 cm2 will be swabbed at each location. A seventh swab will be exposed to the vacuum of space without touching any surfaces as a blank. The eighth swab will remain sealed as a process control. After the EVA the swab kit will be frozen at -80°C and returned to earth at the earliest possible opportunity. The samples will remain frozen until they are thawed for next generation DNA sequencing on earth. We will use amplicon sequencing to identify any bacteria, archaea or fungi present in the samples. If there is enough DNA present, we will use shotgun metagenomic sequencing to further characterize the microbial ecology outside ISS. Preliminary Results: Results from ground-based testing demonstrate that the swab kit is capable of cycling in and out of vacuum without contaminating the swabs. Test subjects, wearing flight-like EVA gloves could remove swabs from the canister and sample discrete locations without inadvertently touching any other surfaces. Several types of bacteria and fungi were collected during these ground tests and survived up to 6 hours at vacuum. This includes non-spore forming bacterial like Staphyloccus capitis that are not traditionally considered extremophiles. Shotgun metagenomic sequencing of the ground-test samples revealed bacteria associated with human skin and airways on the exterior of space suits used during these tests 2 In addition to these results we will present preliminary results from our space-flight samples. We will also present lessons learned from attempting to collect microbiological samples during an EVA and describe how our results will affect planetary protection requirements for future crewed missions.

Aaron B. Regberg↗

NASA NTRSDate not supplied

Revisiting the Soyuz-1 Parachute Failure in the Context of Safety in the Modern Era

The Soyuz‑1 accident remains one of the most consequential parachute related failures in human spaceflight history and provides enduring lessons for modern Entry, Descent, and Landing (EDL) system design. Occurring during the height of the Cold War and the Space Race, the mission unfolded under extraordinary political and schedule pressure as the Soviet Union sought to maintain its early leadership in space achievements following the death of chief designer Sergei Korolev. Despite unresolved propulsion, electrical, and parachute system deficiencies, Soyuz‑1 proceeded to launch and immediately encountered critical inflight anomalies, including a failed solar panel deployment, attitude control issues, and communication dropouts. Upon reentry, a malfunction in the parachute system, driven by a primary main canopy that failed to deploy, and subsequent entanglement of the reserve main canopy with the primary drogue parachute, resulted in insufficient deceleration and the fatal crash of cosmonaut Vladimir Komarov. Subsequent investigations revealed deep rooted cultural and organizational issues within the Soviet space program, including inadequate testing, suppression of dissent, undocumented last minute design changes, and the absence of integrated parachute system verification. More than 200 design flaws were identified after the accident, and firsthand accounts, including those from Yuri Gagarin, highlighted widespread concern prior to launch. Over time, the Soviet program implemented substantial reforms: systematic design corrections, rigorous process documentation, and an extensive series of drop tests that ultimately transformed the Soyuz system into one of the world’s most reliable human-rated return vehicles. This paper examines the technical architecture of the Soyuz‑1 parachute system, reconstructs the likely deployment sequence and failure mechanism, and analyzes the cultural contributors that shaped the accident. The study draws parallels to modern spacecraft parachute development, emphasizing the critical importance of integrated system testing, transparent engineering culture, and continuous hardware surveillance. These lessons remain directly relevant to today’s NASA and Commercial Crew Programs (CCP), where the Government continues to refine its understanding of aggregate risk and strengthen overall astronaut safety in the face of increasingly complex parachute systems.

Aaron L Morris↗

DOE OSTI2022

An in silico method to assess antibody fragment polyreactivity

Antibodies are essential biological research tools and important therapeutic agents, but some exhibit non-specific binding to off-target proteins and other biomolecules. Such polyreactive antibodies compromise screening pipelines, lead to incorrect and irreproducible experimental results, and are generally intractable for clinical development. Here, we design a set of experiments using a diverse naïve synthetic camelid antibody fragment (nanobody) library to enable machine learning models to accurately assess polyreactivity from protein sequence (AUC > 0.8). Moreover, our models provide quantitative scoring metrics that predict the effect of amino acid substitutions on polyreactivity. We experimentally test our models’ performance on three independent nanobody scaffolds, where over 90% of predicted substitutions successfully reduced polyreactivity. Importantly, the models allow us to diminish the polyreactivity of an angiotensin II type I receptor antagonist nanobody, without compromising its functional properties. We provide a companion web-server that offers a straightforward means of predicting polyreactivity and polyreactivity-reducing mutations for any given nanobody sequence.

60 APPLIED LIFE SCIENCES↗

NASA NTRS1989

Gravitational radiation from extreme Kerr black hole

Gravitational radiation induced by a test particle falling into an extreme Kerr black hole was investigated analytically. Assuming the radiation is dominated by the infinite sequence of quasi-normal modes which has the limiting frequency m/(2M), where m is an azimuthal eigenvalue and M is the mass of the black hole, it was found that the radiated energy diverges logarithmically in time. Then the back reaction to the black hole was evaluated by appealing to the energy and angular momentum conservation laws. It was found that the radiation has a tendency to increase the ratio of the angular momentum to mass of the black hole, which is completely different from non-extreme case, while the contribution of the test particle is to decrease it.

Sasaki, Misao↗

NASA NTRS1982

Test and analysis of Celion 3000/PMR-15, graphite/polyimide bonded composite joints: Data report

Standard single lap, double lap and symmetric step lap bonded joints of Celion 3000/PMR-15 graphite/polyimide composite were evaluated. Composite to composite and composite to titanium joints were tested at 116 K (-250 F), 294 K (70 F) and 561 K (550 F). Joint parameters evaluated are lap length, adherend thickness, adherend axial stiffness, lamina stacking sequence and adherend tapering. Advanced joint concepts were examined to establish the change in performance of preformed adherends, scalloped adherends and hybrid systems. The material properties of the high temperature adhesive, designated A7F, used for bonding were established. The bonded joint tests resulted in interlaminar shear or peel failures of the composite and there were very few adhesive failures. Average test results agree with expected performance trends for the various test parameters. Results of finite element analyses and of test/analysis correlations are also presented.

Cushman, J. B.↗

NASA NTRS2023

Effect of Loading Rate on the Open Hole Compression Strength of Quasi-Isotropic Carbon/Epoxy Laminates

This study was undertaken to examine how well the 0.05 inch/min loading rate typically used to develop allowables for open hole compression strength of carbon/epoxy laminates represents structures that may experience a much lower loading rate (such as civil engineering structures) or a much higher loading rate (such as a rocket interstage at liftoff). The Composite Materials Handbook-17 (CMH-17) suggests developing strength allowables at the coupon level (lowest level of the “building block” approach) and using a rigorous statistical analysis to arrive at a strength allowable upon which the structure is to be further designed at higher levels of the building block pyramid. But do these coupon level tests truly represent meaningful stresses at which the composite in the final structure can withstand considering the effects of loading rate? Open hole compression (OHC) strength was chosen for this study since most structures are designed to some damage tolerance criteria and OHC strength testing is rather simple to perform. A quasi-isotropic lay-up was chosen since this is a very common lay-up sequence. The results show that for structures that experience a very small loading rate (0.0005 in/min) the allowables developed at the standard 0.05 in/min loading rate are higher than what the composite can withstand at this lower loading rate. Conversely, at a high loading rate of 10 in/min the strength values found at the standard test rate of 0.05 in/min are slightly lower and not “in family” using the statistics suggested in CMH-17. These results question if standardized test rates are reliable, at least to the fidelity of being “in family” per CMH-17. Some creep rupture tests were also performed to demonstrate that OHC strength values could be lower than the slowest test speed used in this study (0.0005 in/min).

open hole compression strength↗

DOE OSTI2023

De novo design of knotted tandem repeat proteins

De novo protein design methods can create proteins with folds not yet seen in nature. These methods largely focus on optimizing the compatibility between the designed sequence and the intended conformation, without explicit consideration of protein folding pathways. Deeply knotted proteins, whose topologies may introduce substantial barriers to folding, thus represent an interesting test case for protein design. Here we report our attempts to design proteins with trefoil (3 1 ) and pentafoil (5 1 ) knotted topologies. We extended previously described algorithms for tandem repeat protein design in order to construct deeply knotted backbones and matching designed repeat sequences (N = 3 repeats for the trefoil and N = 5 for the pentafoil). We confirmed the intended conformation for the trefoil design by X ray crystallography, and we report here on this protein’s structure, stability, and folding behaviour. The pentafoil design misfolded into an asymmetric structure (despite a 5-fold symmetric sequence); two of the four repeat-repeat units matched the designed backbone while the other two diverged to form local contacts, leading to a trefoil rather than pentafoil knotted topology. Our results also provide insights into the folding of knotted proteins.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS2024

Development Testing and Analysis of the Integrated Gateway-ESPRIT Bipropellant Refuelling System

The Gateway will be humanity’s first space station orbiting the Moon completed by NASA in partnership with ESA and other US and international partners. The Gateway will provide critical support to sustainable human exploration on the moon through the Artemis program. To enable the Lunar Gateway to complete its mission, on orbit refuelling is essential. The ESPRIT Refuelling Module (ERM) will provide the capability to transfer propellants, MMH and MON-3, through the Habitation and Logistics Outpost (HALO) to the Power and Propulsion Element (PPE). The transfer of these propellants carries known risks and hazards. These hazards include overpressure of the propellant lines during priming sequences between modules and during refuelling pause operations. To support the early system development and to mitigate these risks, a collaborative test program between NASA, ESA and Thales Alenia Space was completed at the Thales Alenia Space test facility in Harwell, UK. This test program integrated fluidic breadboards of the ERM, HALO and PPE modules. The objectives of the test program were to demonstrate and characterise critical performance and transient operations, inform refuelling concept of operations and to calibrate and validate numerical models of the refuelling subsystem in EcosimPro. To support the completion of this final objective a detailed model of the integrated breadboard was developed in EcosimPro and key steady-state and transient test cases were simulated. As an industry first, a National Institute of Standards and Technology (NIST) database correlation for Hydrofluoroether (HFE-7000) was used as a mixed oxides of nitrogen (MON-3) simulant with EcosimPro and European Space Propulsion System Simulation (ESPSS) libraries to result in a more accurate analysis for transient phenomenon such as priming. The test and simulation data showed good agreement validating the model for further system analysis as the ERM design progresses.

Refuelling↗

DOE OSTI2025

Nucleic Acid-Based Detection Protease Activity

Proteases include clinically relevant markers for clotting disorders, certain cancers as well as toxins. Assays for protease activity often use designed peptides mimicking natural substrates and detection with colorometric and fluorescence-based detection that is difficult to multiplex without expensive and resource demanding instruments. This work demonstrates detection of proteolytic activity using PCR and sequencing-readable reporter molecules. The assay development focused on binding the constructed peptide-oligonucleotide chimera to immobilized streptavidin. Thrombin, an essential component of the clotting cascade, was used as a model system for testing peptide substrate recognition and release of a designed oligonucleotide for detection. Detection of protease activity was demonstrated in a concentration-dependent manner using MALDI-MS, RT-PCR and DNA sequencing.

Wunschel, David S [Pacific Northwest National Labo↗

DOE OSTI2021

National Virtual Biotechnology Laboratory: Report on Rapid R&D Solutions to the COVID-19 Crisis

With funding from the CARES Act, the U.S Department of Energy (DOE) established the National Virtual Biotechnology Laboratory (NVBL) in March 2020 to address key challenges associated with the COVID-19 crisis. NVBL brought together the broad scientific and technical expertise and resources of DOE’s 17 national laboratories to help tackle medical supply short ages, discover potential drugs to fight the virus, develop and validate COVID-19 testing methods, model disease spread and impact across the nation, and understand virus transport in buildings and the environment. National laboratory resources leveraged for this effort include a suite of world-leading user facilities broadly available to the research community, such as light and neutron sources, nanoscale science research centers, sequencing and biocharacterization facilities, and high-performance computing facilities. Within months, NVBL teams produced innovations in materials and advanced manufacturing that mitigated shortages in test kits and personal protective equipment (PPE), creating nearly 1,000 new jobs. They used DOE’s high-performance computers and light and neutron sources to identify promising candidates for antibodies and antivirals that universities and drug companies are now evaluating. NVBL researchers also developed new diagnostic targets and sample collection approaches, and supported U.S. Food and Drug Administration (FDA), Centers for Disease Control and Prevention (CDC), and U.S. Department of Defense (DoD) efforts to establish national guidelines used in administering millions of tests. Researchers used artificial intelligence and high-performance computing to produce near-real-time data analysis to forecast disease transmission, stress on public health infrastructure, and economic impact, which supported decision-makers at the local, state, and national levels. NVBL teams also studied how to control indoor virus movement to minimize uptake and protect human health. NVBL’s accomplishments demonstrate not only the powerful resource represented by DOE’s national laboratories working together to meet national needs, but also the effectiveness of the integrated NVBL framework for rapidly responding to emergencies with research and development (R&D) solutions. As the fight against COVID continues, sustained efforts are needed to confront this pandemic as well as future threats. Examples include: 1) Establishing “supply chains on demand” to meet emergency production needs by leveraging the materials and manufacturing expertise of DOE national laboratories and developing advances in electronics, sensing, robotics, and automation capabilities; 2) Improving the speed and robustness of drug discovery by integrating experimental platforms with DOE’s computational and experimental user facilities, which provide unique resources to support the discovery of high-potential therapeutic agents; 3) Protecting public, environmental, and animal health by developing new testing protocols and instrumentation adaptable to diverse sample types (both physiological and environmental) to quickly detect a wide range of pathogens and monitor other biorisks; 4) Supporting near-real-time data needs of decision-makers at the local, regional, state, and national levels by advancing data curation, analysis, and modeling using artificial intelligence and new data science tools for managing and evaluating large diverse datasets; 5) Harnessing DOE’s expertise in environmental modeling to design rooms and air handling for offices, classrooms, restaurants, and other structures to minimize biorisk transmissions. Going forward, NVBL is poised to apply the unique capabilities and expertise of the national laboratory complex to future national and international emergencies, both natural and engineered. Through this framework, the Office of Science will continue to be an integral component of agency wide efforts to prepare for and respond to biorisks and other crises.

42 ENGINEERING↗

NASA NTRS1979

Fabrication and testing of composite ring specimens

The tooling and techniques used in the fabrication of composite laminate tubes of any desired ply orientation and stacking sequence are described along with techniques for cutting ring specimens under internal pressure. The method consists of laying up the tube on a central circular mandrel, and by means of internal pressure, expanding the prepreg tube against the cavity wall of an external mold tool, which forms the geometric curing envelope for the tube. Tube quality is assessed by laminate wall thickness measurement, by hoop strength measurement on rings cut from the ends of the tube, and by ultrasonic inspection.

Liber, T.↗

DOE OSTI2021

Zymomonas diversity and potential for biofuel production

Abstract Background Zymomonas mobilis is an aerotolerant α-proteobacterium, which has been genetically engineered for industrial purposes for decades. However, a comprehensive comparison of existing strains on the genomic level in conjunction with phenotype analysis has yet to be carried out. We here performed whole-genome comparison of 17 strains including nine that were sequenced in this study. We then compared 15 available Zymomonas strains for their natural abilities to perform under conditions relevant to biofuel synthesis. We tested their growth in anaerobic rich media, as well as growth, ethanol production and xylose utilization in lignocellulosic hydrolysate. We additionally compared their tolerance to isobutanol, flocculation characteristics, and ability to uptake foreign DNA by electroporation and conjugation. Results Using clustering based on 99% average nucleotide identity (ANI), we classified 12 strains into four clusters based on sequence similarity, while five strains did not cluster with any other strain. Strains belonging to the same 99% ANI cluster showed similar performance while significant variation was observed between the clusters. Overall, conjugation and electroporation efficiencies were poor across all strains, which was consistent with our finding of coding potential for several DNA defense mechanisms, such as CRISPR and restriction–modification systems, across all genomes. We found that strain ATCC31821 (ZM4) had a more diverse plasmid profile than other strains, possibly leading to the unique phenotypes observed for this strain. ZM4 also showed the highest growth of any strain in both laboratory media and lignocellulosic hydrolysate and was among the top 3 strains for isobutanol tolerance and electroporation and conjugation efficiency. Conclusions Our findings suggest that strain ZM4 has a unique combination of genetic and phenotypic traits that are beneficial for biofuel production and propose investing future efforts in further engineering of ZM4 for industrial purposes rather than exploring new Zymomonas isolates.

09 BIOMASS FUELS↗

NASA NTRS2002

Mode shift of an inhaled aerosol bolus is correlated with flow sequencing in the human lung

We studied the effects on aerosol bolus inhalations of small changes in convective inhomogeneity induced by posture change from upright to supine in nine normal subjects. Vital capacity single-breath nitrogen washout tests were used to determine ventilatory inhomogeneity change between postures. Relative to upright, supine phase III slope was increased 33 +/- 11% (mean +/- SE, P < 0.05) and phase IV height increased 25 +/- 11% (P < 0.05), consistent with an increase in convective inhomogeneity likely due to increases in flow sequencing. Subjects also performed 0.5-microm-particle bolus inhalations to penetration volumes (V(p)) between 150 and 1,200 ml during a standardized inhalation from residual volume to 1 liter above upright functional residual capacity. Mode shift (MS) in supine posture was more mouthward than upright at all V(p), changing by 11.6 ml at V(p) = 150 ml (P < 0.05) and 38.4 ml at V(p) = 1,200 ml (P < 0.05). MS and phase III slope changes correlated positively at deeper V(p). Deposition did not change at any V(p), suggesting that deposition did not cause the MS change. We propose that the MS change results from increased sequencing in supine vs. upright posture.

NASA Discipline Cardiopulmonary↗