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At least 181 records · Page 10

WetLab-2: Tools for Conducting On-Orbit Quantitative Real-Time Gene Expression Analysis on ISS

The objective of NASA Ames Research Centers WetLab-2 Project is to place on the ISS a research platform capable of conducting gene expression analysis via quantitative real-time PCR (qRT-PCR) of biological specimens sampled or cultured on orbit. The project has selected a Commercial-Off-The-Shelf (COTS) qRT-PCR system, the Cepheid SmartCycler and will fly it in its COTS configuration. The SmartCycler has a number of advantages including modular design (16 independent PCR modules), low power consumption, rapid ramp times and the ability to detect up to four separate fluorescent channels at one time enabling multiplex assays that can be used for normalization and to study multiple genes of interest in each module. The team is currently working with Cepheid to enable the downlink of data from the ISS to the ground and provide uplink capabilities for programming, commanding, monitoring, and instrument maintenance. The project has adapted commercial technology to design a module that can lyse cells and extract RNA of sufficient quality and quantity for use in qRT-PCR reactions while using a housekeeping gene to normalize RNA concentration and integrity. The WetLab-2 system is capable of processing multiple sample types ranging from microbial cultures to animal tissues dissected on-orbit. The ability to conduct qRT-PCR on-orbit eliminates the confounding effects on gene expression of reentry stresses and shock acting on live cells and organisms or the concern of RNA degradation of fixed samples. The system can be used to validate terrestrial analyses of samples returned from ISS by providing on-orbit gene expression benchmarking prior to sample return. The ability to get on orbit data will provide investigators with the opportunity to adjust experiment parameters for subsequent trials based on the real-time data analysis without need for sample return and re-flight. Researchers will also be able to sample multigenerational changes in organisms. Finally, the system can be used for analysis of air, surface, water, and clinical samples to monitor environmental contaminants and crew health. The verification flight of the instrument is scheduled to launch on SpaceX-7 in June 2015.

quantitative PCR↗

Wetlab-2 - Quantitative PCR Tools for Spaceflight Studies of Gene Expression Aboard the International Space Station

Wetlab-2 is a research platform for conducting real-time quantitative gene expression analysis aboard the International Space Station. The system enables spaceflight genomic studies involving a wide variety of biospecimen types in the unique microgravity environment of space. Currently, gene expression analyses of space flown biospecimens must be conducted post flight after living cultures or frozen or chemically fixed samples are returned to Earth from the space station. Post-flight analysis is limited for several reasons. First, changes in gene expression can be transient, changing over a timescale of minutes. The delay between sampling on Earth can range from days to months, and RNA may degrade during this period of time, even in fixed or frozen samples. Second, living organisms that return to Earth may quickly re-adapt to terrestrial conditions. Third, forces exerted on samples during reentry and return to Earth may affect results. Lastly, follow up experiments designed in response to post-flight results must wait for a new flight opportunity to be tested.

ISS research platform↗

On-Orbit Quantitative Real-Time Gene Expression Analysis Using the Wetlab-2 System

NASA Ames Research Center's WetLab-2 Project enables on-orbit quantitative Reverse Transcriptase PCR (qRT-PCR) analysis without the need for sample return. The WetLab-2 system is capable of processing sample types ranging from microbial cultures to animal tissues dissected on-orbit. The project developed a RNA preparation module that can lyse cells and extract RNA of sufficient quality and quantity for use as templates in qRT-PCR reactions. Our protocol has the advantage of using non-toxic chemicals and does not require alcohols or other organics. The resulting RNA is dispensed into reaction tubes that contain all lyophilized reagents needed to perform qRT-PCR reactions. System operations require simple and limited crew actions including syringe pushes, valve turns and pipette dispenses. The project selected the Cepheid SmartCycler (TradeMark), a Commercial-Off-The-Shelf (COTS) qRT-PCR unit, because of its advantages including rugged modular design, low power consumption, rapid thermal ramp times and four-color multiplex detection. Single tube multiplex assays can be used to normalize for RNA concentration and integrity, and to study multiple genes of interest in each module. The WetLab-2 system can downlink data from the ISS to the ground after a completed run and uplink new thermal cycling programs. The ability to conduct qRT-PCR and generate results on-orbit is an important step towards utilizing the ISS as a National Laboratory facility. Specifically, the ability to get on-orbit data will provide investigators with the opportunity to adjust experimental parameters in real time without the need for sample return and re-flight. On orbit gene expression analysis can also eliminate the confounding effects on gene expression of reentry stresses and shock acting on live cells and organisms or the concern of RNA degradation of fixed samples and provide on-orbit gene expression benchmarking prior to sample return. Finally, the system can also be used for analysis of air, surface, water, and clinical samples to monitor environmental pathogens and crew health. The validation flight of the WetLab-2 system using E. coli bacteria and mouse liver launched on SpaceX-7 in June 2015 and will remain on the ISS National Laboratory.

ISS tools↗

Inception of a Spaceflight-specific Mouse to Human Expression Profiling Translation Model

Rodents are foundational model organisms often utilized due to their seemingly analogous morphologies and biological responses to humans. However, recent studies have demonstrated that murine model data are limited in their applicability, particularly in inflammatory disease. In space studies, accurately predicting human response from mouse data is critical due to extreme limiting factors in both rodent and human spaceflight research. With successful prediction, spaceflight ailments can be predicted and prevented while respecting the constraints of the spaceflight industry and minimizing danger to humans. To do so, novel methodologies must be developed that predict human response from murine data after considering biological differences between rodents and humans in spaceflight. After considering terrestrial models, we determined that a spaceflight-based expression profiting translation tool should be created to accurately capture predictions of human gene expression in spaceflight from mouse data. To prepare to build this model, we organized known human spaceflight risks, chose analog human diseases as training data categories, then identified existing RNASeq disease datasets from GEO as potential training data. In addition, we classified existing Genelab mouse differential gene expression datasets for use as experimental data.

Translation↗

Investigation of a lineament expressed in an oblique Apollo 9 photograph

The author has identified the following significant results. A linear topographic feature, referred to as the New York Mountains lineament, was recognized in an oblique Apollo 9 photograph to extend from the Providence Mountains of California to near Lake Mead, Arizona. In subsequent vertical ERTS-1 imagery this feature was found to have vague and indistinct expression. A study was conducted to determine the possible geologic origin(s) of the lineament and to explain its anomalous expression in the Apollo 9 photograph. The results suggest that the apparent expression of the lineament is due to a combination of the oblique view of the Apollo photograph, low sun angle illumination of southeast facing slopes, shadowing of northwest facing slopes, and linear snow line along the southeastern flank of the New York Mountains. No geologic or structural causes for the lineament have been found.

Liggett, M. A.↗

Wide range stress intensity factor expressions for ASTM E 399 standard fracture toughness specimens

For each of the two types of specimens, bend and compact, described previously for plane strain fracture toughness of materials, E 399, a polynominal expression is given for calculation of the stress intensity factor, K, from the applied force, P, and the specimen dimensions. It is explicitly stated, however, that these expressions should not be used outside the range of relative crack length, a/W, from 0.45 to 0.55. While this range is sufficient for the purpose of E 399, the same specimen types are often used for other purposes over a much wider range of a/W; for example, in the study of fatigue crack growth. Expressions are presented which are at least as accurate as those in E 399-74, and which cover much wider ranges of a/W: for the three-point bend specimen from 0 to 1; and for the compact specimen from 0.2 to 1. The range has to be restricted for the compact specimen because of the proximity of the loading pin holes to the crackline, which causes the stress intensity factor to be sensitive to small variations in dimensions when a/W is small. This is a penalty inherently associated with the compactness of the specimen.

Srawley, J. E.↗

A simple expression for kilometric radiation growth rates and analytical applications

A simple expression is derived for the linear growth rate of R-X mode, auroral kilometric radiation under the assumptions of cold plasma wave dispersion and (omega sub pe)-squared is much less than (Omega sub e)-squared, where omega sub pe and Omega sub e are the electron plasma frequency and gyrofrequency, respectively. The result is valid for all wave normal angles and all frequencies above the right-hand cutoff frequency. With this expression, it is not necessary to evaluate the full dispersion tensor, and variations of the growth rate with plasma parameters are easily obtained. Applying the new expression, it is found that kilometric radiation growth rates are strongly dependent upon the magnitude of the magnetic field-aligned potential difference, and that significant growth of radiation at harmonics of the fundamental is unlikely in regions where (omega sub pe)-squared is much less than (Omega sub e)-squared.

Lyons, L. R.↗

A new world survey expression for cosmic ray vertical intensity vs. depth in standard rock

The cosmic ray data on vertical intensity versus depth below 10 to the 5th power g sq cm is fitted to a 5 parameter empirical formula to give an analytical expression for interpretation of muon fluxes in underground measurements. This expression updates earlier published results and complements the more precise curves obtained by numerical integration or Monte Carlo techniques in which the fit is made to an energy spectrum at the top of the atmosphere. The expression is valid in the transitional region where neutrino induced muons begin to be important, as well as at great depths where this component becomes dominant.

Crouch, M.↗

The effects of expressivity and flight task on cockpit communication and resource management

The results of an investigation to develop a methodology for evaluating crew communication behavior on the flight deck and a flight simulator experiment to test the effects of crew member expressivity, as measured by the Personal Attributes Questionnarie, and flight task on crew communication and flight performance are discussed. A methodology for coding and assessing flight crew communication behavior as well as a model for predicting that behavior is advanced. Although not enough crews were found to provide valid statistical tests, the results of the study tend to indicate that crews in which the captain has high expressivity perform better than those whose captain is low in expressivity. There appears to be a strong interaction between captains and first officers along the level of command dimension of communication. The PAQ appears to identify those pilots who offer disagreements and inititate new subjects for discussion.

Jensen, R. S.↗

An exact, closed-form expression of the integral chord-length distribution for the calculation of single-event upsets induced by cosmic rays

This paper presents a derivation of an exact closed-form expression of the integral chord-length distribution for the calculation of single-event upsets (SEUs) in an electronic memory cell, caused by cosmic rays. Results computed for two rectangular parallelepipeds using this exact expression are compared with those computed with Bradford's (1979) semiexact expression of C(x). It is found that the values of C(x) are identical for x equal or smaller than b but are vastly different for x greater than b. Moreover, while C(x) of Bradford gives reasonably accurate values of SEU rate for certain sets of computational parameters, it gives values more than 10 times larger than the correct values for other sets of parameters.

Luke, Keung L.↗

Exact closed-form expressions for the performance of the split-symbol moments estimator of signal-to-noise ratio

Previously, the performance of the split-symbol moments estimator (SSME) of signal-to-noise ratio (SNR) has been evaluated by means of approximate expressions for the estimator mean and variance. These are asymptotic formulas in the sense that they become accurate as the number of estimator samples gets large. Here, exact closed-form expressions are obtained for the same quantities. These expressions confirm the accuracy of the previously derived asymptotic results, and, unlike the asymptotic formulas, they are useful even when the number of samples is small. It is also shown that the conventional split-symbol estimator can be trivially scaled to form a signal-to-noise ratio estimator which is precisely unbiased (as long as the estimate is based on more than two split-symbols).

Dolinar, S.↗

Global Expression for Representing Diatomic Potential-Energy Curves

A three-parameter expression that gives an accurate fit to diatomic potential curves over the entire range of separation for charge transfers between 0 and 1. It is based on a generalization of the universal binding-energy relation of Smith et al. (1989) with a modification that describes the crossover from a partially ionic state to the neutral state at large separations. The expression is tested by comparison with first-principles calculations of the potential curves ranging from covalently bonded to ionically bonded. The expression is also used to calculate spectroscopic constants form a curve fit to the first-principles curves. A comparison is made with experimental values of the spectroscopic constants.

Ferrante, John↗

Analytical expressions for radiative properties of planar Rayleigh scattering media, including polarization contributions

The objective of this paper is to provide a convenient and fast way for computing the radiative properties of planar Rayleigh scattering media. Analytical expressions are developed for the three molecular atmospheric functions which are required in remote sensing: the atmospheric reflectance, the transmission function, and the spherical albedo. The expressions are adjusted by using accurate computations performed with successive orders of a scattering code. The accuracy of the code is first checked by using previously published tabulations. The required accuracy of 0.001 in the reflectance unit is achieved by numerical adjustments. The contribution of polarization is considered implicitly. The expressions are shown to be valid for a fairly large range of observational conditions.

Vermote, E.↗

Optimal evaluation of array expressions on massively parallel machines

We investigate the problem of evaluating FORTRAN 90 style array expressions on massively parallel distributed-memory machines. On such machines, an elementwise operation can be performed in constant time for arrays whose corresponding elements are in the same processor. If the arrays are not aligned in this manner, the cost of aligning them is part of the cost of evaluating the expression. The choice of where to perform the operation then affects this cost. We present algorithms based on dynamic programming to solve this problem efficiently for a wide variety of interconnection schemes, including multidimensional grids and rings, hypercubes, and fat-trees. We also consider expressions containing operations that change the shape of the arrays, and show that our approach extends naturally to handle this case.

Chatterjee, Siddhartha↗

Express Payload Project - A new method for rapid access to Space Station Freedom

The deployment and permanent operation of Space Station Freedom will enable researchers to enter a new era in the 21st century, in which continuous on-orbit experimentation and observation become routine. In support of this objective, the Space Station Freedom Program Office has initiated the Express Payload Project. The fundamental project goal is to reduce the marginal cost associated with small payload development, integration, and operation. This is to be accomplished by developing small payload accommodations hardware and a new streamlined small payload integration process. Standardization of small payload interfaces, certification of small payload containers, and increased payload developer responsibility for mission success are key aspects of the Express Payload Project. As the project progresses, the principles will be applied to both pressurized payloads flown inside the station laboratories and unpressurized payloads attached to the station external structures. The increased access to space afforded by Space Station Freedom and the Express Payload Project has the potential to significantly expand the scope, magnitude, and success of future research in the microgravity environment.

Uhran, Mark L.↗

Central coast designs: The Eightball Express. Taking off with convention, cruising with improvements and landing with absolute success

The airline industry is very competitive, resulting in most U.S. and many international airlines being unprofitable. Because of this competition the airlines have been engaging in fare wars (which reduce revenue generated by transporting passengers) while inflation has increased. This situation of course is not developing revenue for the airlines. To revive the airlines to profitability, the difference between revenue received and airline operational cost must be improved. To solve these extreme conditions, the Eightball Express was designed with the main philosophy of developing an aircraft with a low direct operating cost and acquisition cost. Central Coast Designs' (CCD) aircraft utilizes primarily aluminum in the structure to minimize manufacturing cost, supercritical airfoil sections to minimize drag, and fuel efficient engines to minimize fuel burn. Furthermore, the aircraft was designed using Total Quality Management and Integrated Product Development to minimize development and manufacturing costs. Using these primary cost reduction techniques, the Eightball Express was designed to meet the Lockheed/AIAA Request for Proposal (RFP) requirements of a low cost, 153 passenger, 3000 nm. range transport. The Eightball Express is able to takeoff on less than a 7000 ft. runway, cruise at Mach 0.82 at an altitude of 36,000 ft. for a range of 3,000 nm., and lands on a 5,000 ft. runway. lt is able to perform this mission at a direct operating cost of 3.51 cents/available seat mile in 1992 dollars while the acquisition cost is only $28 million in 1992 dollars. By utilizing and improving on proven technologies, CCD has produced an efficient low cost commercial transport for the future.

Davis, Ryan Edwin↗

Protein expression in Arabidopsis thaliana after chronic clinorotation

Soluble protein expression in Arabidopsis thaliana L. (Heynh.) leaf and stem tissue was examined after chronic clinorotation. Seeds of Arabidopsis were germinated and plants grown to maturity on horizontal or vertical slow-rotating clinostats (1 rpm) or in stationary vertical control units. Total soluble proteins and in vivo-labeled soluble proteins isolated from these plants were analyzed by two-dimensional sodium doedocyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and subsequent fluorography. Visual and computer analysis of the resulting protein patterns showed no significant differences in either total protein expression or in active protein synthesis between horizontal clinorotation and vertical controls in the Arabidopsis leaf and stem tissue. These results show chronic clinorotation does not cause gross changes in protein expression in Arabidopsis.

Piastuch, William C.↗

ISS External Payload Accommodations (EXPRESS pallet)

The 'expedite the process of experiments to Space Station' (EXPRESS) pallet which is attached to the DS3 truss segment of the International Space Station (ISS) via the payload attach structure is illustrated. The EXPRESS pallet constitutes the primary ISS external payload carrier. Each pallet carries six robotically replaceable payload adapters which are capable of containing one or more payloads. The following aspects of the EXPRESS program and pallet are illustrated: the concept drivers; the physical integration; the installation and in-orbit replacement; and the experiments to be implemented. The program status is summarized.

Harvey L. Shelton↗