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At least 235 records · Page 13

Smectites versus palagonites in Mars soil: Evidence from simulations of Viking biology labeled release experiments

The results of an experimental comparison between palagonites and a smectite (montmorillonite) in the simulation of the Viking Biology Labeled Release (LR) experiment and conclusions regarding their suitability as MarSAMs are reproved. It was found that palagonites do not cause formate decomposition and C-14 release in their natural form or after acidification and thus cannot be a completely satisfactory analog to the Mars soil studied by Viking.

Banin, A.↗

Edge Labels for Multilayer Circuit Boards

Quick visual inspection identifies out-of-place layers. Small copper pads or other labels on layers present regular pattern if multilayer circuit board properly assembled and irregular pattern if one or more layers in wrong place. New method faster, cheaper, and more reliable than earlier methods.

Rossi, P. J.↗

Metabolism of Nonessential N15-Labeled Amino Acids and the Measurement of Human Whole-Body Protein Synthesis Rates

Eight N-15 labeled nonessential amino acids plus (15)NH4Cl were administered over a 10 h period to four healthy adult males using a primed-constant dosage regimen. The amount of N-15 excreted in the urine and the urinary ammonia, hippuric acid, and plasma alanine N-15 enrichments were measured. There was a high degree of consistency across subjects in the ordering of the nine compounds based on the fraction of N-15 excreted (Kendall coefficient of concordance W = 0.83, P is less than 0.01). Protein synthesis rates were calculated from the urinary ammonia plateau enrichment and the cumulative excretion of N-15. Glycine was one of the few amino acids that gave similar values by both methods.

Stein, T. P.↗

Metabolism of nonessential N-15-labeled amino acids and the measurement of human whole-body protein synthesis rates

Eight N-15-labeled nonessential amino acids plus (N-15)H4Cl were administered over a 10-h period to four healthy adult males using a primed-constant dosage regimen. The amount of N-15 excreted in the urine and the urinary ammonia, hippuric acid, and plasma alanine N-15 enrichments were measured. There was a high degree of consistency across subjects in the ordering of the nine compounds based on the fraction of N-15 excreted.

Stein, T. P.↗

Parallel algorithms for geometric connected component labeling on a hypercube multiprocessor

Different algorithms for the geometric connected component labeling (GCCL) problem are defined each of which involves d stages of message passing, for a d-dimensional hypercube. The major idea is that in each stage a hypercube multiprocessor increases its knowledge of domain. The algorithms under consideration include the QUAD algorithm for small number of processors and the Overlap Quad algorithm for large number of processors, subject to the locality of the connected sets. These algorithms differ in their run time, memory requirements, and message complexity. They were implemented on an Intel iPSC2/D4/MX hypercube.

Belkhale, K. P.↗

Analyst variability in labeling of unsupervised classifications

Analyst variability in the labeling of unsupervised classifications is tested for Landsat 5 Thematic Mapper image products covering two test sites in southern California. The accuracy of results are tested using samples from a photo interpreted base map of the area. The significance of differences between analysts is indicated by comparing Kappa statistics derived from error matrices. Analyst variability is found to be statistically significant in most cases. Certain analysts provided consistently better results for a given study area or degree of training. This work demonstrates the potential influence of analyst bias on what would otherwise seem to be a fairly objective method and suggests that controls for this subjectivity should be factored into experimental designs.

Mcgwire, Kenneth C.↗

Nucleotide-Protectable Labeling of Sulfhydryl Groups in Subunit I of the ATPhase from Halobacterium Saccharovorum

A membrane-bound ATPase from the archaebacterium Halobacterium saccharovorum is inhibited by N-ethyl-maleimide in a nucleotide-protectable manner. When the enzyme was incubated with N-[C-14]jethylmaleimide, the bulk of radioactivity was as- sociated with the 87,000-Da subunit (subunit 1). ATP, ADP, or AMP reduced incorporation of the inhibitor. No charge shift of subunit I was detected following labeling with N-ethylmaleimide, indicating an electroneutral reaction. The results are consistent with the selective modification of sulfhydryl groups in subunit I at or near the catalytic site and are further evidence of a resemblance between this archaebacterial ATPase and the vacuolar-type ATPases.

Sulzner, Michael↗

Carbon Nanotube Nanoelectrode Array as an Electronic Chip for Ultrasensitive Label-free DNA Detection

A reliable nanoelectrode array based on vertically aligned multi-walled carbon nanotubes (MWNTs) embedded in SiO2 is used for ultrasensitive DNA detection. Characteristic nanoelectrode behavior is observed using low-density MWNT arrays for measuring both bulk and surface immobilized redox species such as K4Fe(CN)6 and ferrocene derivatives. The open-end of MWNTs are found to present similar properties as graphite edge-plane electrodes with wide potential window, flexible chemical functionalities, and good biocompatibility. BRCA1 related oligonucleotide probes with 18 bp are selectively functionalized at the open ends of the nanotube array and specifically hybridized with oligonucleotide targets incorporated with a polyG tag. The guanine groups are employed as the signal moieties in the electrochemical measurements. R(bpy)(sup 2+, sub 3) mediator is used to further amplify the guanine oxidation signal. The hybridization of sub-attomoles of DNA targets is detected electrochemically by combining the MWNT nanoelectrode array with the R(bpy)(sup 2+, sub 3) amplification mechanism. This technique was employed for direct electrochemical detection of label-free PCR amplicon from a healthy donor through specific hybridization with the BRCA1 probe. The detection limit is estimated to be less than 1000 DNA molecules since abundant guanine bases in the PCR amplicon provides a large signal. This system provides a general platform for rapid molecular diagnostics in applications requiring ultrahigh sensitivity, high-degree of miniaturization, and simple sample preparation, and low-cost operation.

Li, Jun↗

[13C]-Specific labeling of 8-2' linked (-)-cis-blechnic, (-)-trans-blechnic and (-)-brainic acids in the fern Blechnum spicant

In vivo administration experiments using stable (13C) and radio (14C) labeled precursors established that the optically active 8-2' linked lignans, (-)-cis-blechnic, (-)-trans-blechnic and (-)-trans-brainic acids, were directly derived from L-phenylalanine, cinnamate, and p-coumarate but not either from tyrosine or acetate. The radiochemical time course data suggest that the initial coupling product is (-)-cis-blechnic acid, which is then apparently converted into both (-)-trans-blechnic and (-)-trans-brainic acids in vivo. These findings provide additional evidence for vascular plant proteins engendering distinct but specific phenolic radical-radical coupling modes, i.e., for full control over phenylpropanoid coupling in vivo, whether stereoselective or regiospecific.

NASA Discipline Plant Biology↗

Rationally designed fluorescently labeled sulfate-binding protein mutants: evaluation in the development of a sensing system for sulfate

Periplasmic binding proteins from E. coli undergo large conformational changes upon binding their respective ligands. By attaching a fluorescent probe at rationally selected unique sites on the protein, these conformational changes in the protein can be monitored by measuring the changes in fluorescence intensity of the probe which allow the development of reagentless sensing systems for their corresponding ligands. In this work, we evaluated several sites on bacterial periplasmic sulfate-binding protein (SBP) for attachment of a fluorescent probe and rationally designed a reagentless sensing system for sulfate. Eight different mutants of SBP were prepared by employing the polymerase chain reaction (PCR) to introduce a unique cysteine residue at a specific location on the protein. The sites Gly55, Ser90, Ser129, Ala140, Leu145, Ser171, Val181, and Gly186 were chosen for mutagenesis by studying the three-dimensional X-ray crystal structure of SBP. An environment-sensitive fluorescent probe (MDCC) was then attached site-specifically to the protein through the sulfhydryl group of the unique cysteine residue introduced. Each fluorescent probe-conjugated SBP mutant was characterized in terms of its fluorescence properties and Ser171 was determined to be the best site for the attachment of the fluorescent probe that would allow for the development of a reagentless sensing system for sulfate. Three different environment-sensitive fluorescent probes (1,5-IAEDANS, MDCC, and acylodan) were studied with the SBP171 mutant protein. A calibration curve for sulfate was constructed using the labeled protein and relating the change in the fluorescence intensity with the amount of sulfate present in the sample. The detection limit for sulfate was found to be in the submicromolar range using this system. The selectivity of the sensing system was demonstrated by evaluating its response to other anions. A fast and selective sensing system with detection limits for sulfate in the submicromolar range was developed. Copyright 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 78: 517-526, 2002.

Evaluation Studies↗

A method for the 32P labeling of peptides or peptide nucleic acid oligomers

A novel approach to the radioactive labeling of peptides and PNA oligomers is described. It is based on the conjugation of a deoxynucleoside 3'-phosphate with the terminal amine of the substrate, followed by phosphorylation of the 5'-hydroxyl group of the nucleotide using T4 polynucleotide kinase and [gamma-32P]ATP.

Non-NASA Center↗

Effect of bolus fluid intake on energy expenditure values as determined by the doubly labeled water method

The doubly labeled water (DLW, 2H(2)18O) method is a highly accurate method for measuring energy expenditure (EE). A possible source of error is bolus fluid intake before body water sampling. If there is bolus fluid intake immediately before body water sampling, the saliva may reflect the ingested water disproportionately, because the ingested water may not have had time to mix fully with the body water pool. To ascertain the magnitude of this problem, EE was measured over a 5-day period by the DLW method. Six subjects were dosed with 2H2(18)O. After the reference salivas for the two-point determination were obtained, subjects drank water (700-1,000 ml), and serial saliva samples were collected for the next 3 h. Expressing the postbolus saliva enrichments as a percentage of the prebolus value, we found 1) a minimum in the saliva isotopic enrichments were reached at approximately 30 min with the minimum for 2H (95.48 +/- 0.43%) being significantly lower than the minimum for 18O (97.55 +/- 0.44, P less than 0.05) and 2) EE values calculated using the postbolus isotopic enrichments are appreciably higher (19.9 +/- 7.5%) than the prebolus reference values. In conclusion, it is not advisable to collect saliva samples for DLW measurements within approximately 1 h of bolus fluid intake.

NASA Discipline Musculoskeletal↗

Carbon-13 Labeling Used to Probe Cure and Degradation Reactions of High- Temperature Polymers

High-temperature, crosslinked polyimides are typically insoluble, intractible materials. Consequently, in these systems it has been difficult to follow high-temperature curing or long-term degradation reactions on a molecular level. Selective labeling of the polymers with carbon-13, coupled with solid nuclear magnetic resonance spectrometry (NMR), enables these reactions to be followed. We successfully employed this technique to provide insight into both curing and degradation reactions of PMR-15, a polymer matrix resin used extensively in aircraft engine applications.

Meador, Mary Ann B.↗

Enhanced Labeling Techniques to Study the Cytoskeleton During Root Growth and Gravitropism

Gravity effects the growth and development of all living organisms. One of the most obvious manifestations of gravity's effects on biological systems lies in the ability of plants to direct their growth along a path that is dictated by the gravity vector (called gravitropism). When positioned horizontally, in florescence stems and hypocotyls in dicots, and pulvini in monocots, respond by bending upward whereas roots typically bend downward. Gravitropism allows plants to readjust their growth to maximize light absorption for photosynthesis and to more efficiently acquire water and nutrients form the soil. Despite its significance for plant survival, there are still major gaps in understanding the cellular and molecular processes by which plants respond to gravity. The major aim of this proposal was to develop improved fluorescence labeling techniques to aid in understanding how the cytoskeleton modulated plant responses to gravity.

Blancaflor, Elison B.↗

LC/MS Method for the Determination of Stable Isotope Labeled Promethazine in Human Plasma

Promethazine (PMZ) is taken by astronauts orally (PO), intramuscularly (IM) or rectally (PR) for space motion sickness. LC/MS method was developed with off-line solid phase extraction to measure plasma concentrations of PMZ given as stable isotope-labeled (SIL) formulations by the three different routes of administration simultaneously. Samples (0.5ml) were loaded on to Waters Oasis HLB co-polymer cartridges and eluted with 1.0 mL methanol. HPLC separation of the eluted sample was performed using an Agilent Zorbax SB-CN column (50 x 2.1 mm) at a flow rate of 0.2 mL/min for 6 min. Acetonitrile/ ammonium acetate (30 mM) in water (3:2, v/v), pH 5.6 plus or minus 0.1, was used as the mobile phase for separation. Concentrations of PMZ, PMZ-d4 and PMZ-d7 and chlorpromazine (internal standard) were determined using a Micromass ZMD single quadrupole mass spectrometer with Electrospray Ionization (ESI). ESI mass spectra were acquired in positive ion mode with selected ion monitoring of [M+ H]dot plus. The method is rapid, reproducible and the assay specific parameters are listed in a table. A novel, sensitive and specific method for the measurement of PMZ and SIL PMZ in human plasma is reported.

Zuwei, Wang↗

Fluorescent Quantum Dots for Biological Labeling

Fluorescent semiconductor quantum dots that can serve as "on/off" labels for bacteria and other living cells are undergoing development. The "on/off" characterization of these quantum dots refers to the fact that, when properly designed and manufactured, they do not fluoresce until and unless they come into contact with viable cells of biological species that one seeks to detect. In comparison with prior fluorescence-based means of detecting biological species, fluorescent quantum dots show promise for greater speed, less complexity, greater sensitivity, and greater selectivity for species of interest. There are numerous potential applications in medicine, environmental monitoring, and detection of bioterrorism.

McDonald, Gene↗

Method of Stamping Surface-Enhance Raman Spectroscopy for Label-Free, Multiplexed, Molecular Sensing and Imaging

The present disclosure relates the use of a stamping surface enhanced Raman scattering (S-SERS) technique with nanoporous gold disk (NPGD) plasmonic substrates to produce a label-free, multiplexed molecular sensing and imaging technique. A NPGD SERS substrate is stamped onto a surface containing one or more target molecules, followed by SERS measurement of the target molecules located between the surface and SERS substrate. The target molecules may be deposited on the surface, which may be a carrier substrate such as polydimethylsiloxane (PDMS).

Shih, Wei-Chuan↗

An Automated Approach to Labelling Datasets in Earth Science Publications

NASA Data Active Archive Centers, orDAACs, ingest, store, and distribute dataacquired from satellites, ground systems as well asreanalysis models. Many authors use this datain their research. However, most of the datasets usedin Earth Science Publications are not citedcorrectly or not cited at all. Thus, there is no directlink between the datasets used and thescientific publications which reference them. Thisleads to issues with reproducibility of theresults, attribution of the research results, anddiscovery of new datasets. This project began byexploring various methods of automatically labellingGoddard Earth Sciences Data andInformation Services Center (GES DISC) datasets usingSupervised Machine Learning and EarthData Search Common Metadata Repository (CMR) queries.The ultimate goal was to create alibrary of citations that utilized automated citationlabeling to directly link the researchpublications to the data they use. Supervised MachineLearning approaches struggled due to thelimited amount of labelled training data to learnfrom. Increasing the volume of training data isdifficult as it requires subject matter experts todevote time to manually reviewing journalarticles and determining the datasets used. The CMRqueries were inconsistent because theunderlying metadata is continuously being updated.Thus, it is hard to generalize theeffectiveness of the CMR results as they are dependenton the internal state of CMR. Theseapproaches helped inform the decision to transitionthe project into using a Knowledge Graph.Another key aspect of this project focused on theautomated extraction of features (platform,instrument, variables, etc) and explicit citationsfrom within Earth Science Publications. Theseautomated extractions were used to classify researchpapers based on their platform/instrumentcouples. This information was input into the CitationManagement System for GES DISC. Theseplatform/instrument couples also provide an additionalfacet that can be searched on the GESDISC website.

Edward Jahoda↗