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Methods for determining the genetic affinity of microorganisms and viruses

Selecting which sub-sequences in a database of nucleic acid such as 16S rRNA are highly characteristic of particular groupings of bacteria, microorganisms, fungi, etc. on a substantially phylogenetic tree. Also applicable to viruses comprising viral genomic RNA or DNA. A catalogue of highly characteristic sequences identified by this method is assembled to establish the genetic identity of an unknown organism. The characteristic sequences are used to design nucleic acid hybridization probes that include the characteristic sequence or its complement, or are derived from one or more characteristic sequences. A plurality of these characteristic sequences is used in hybridization to determine the phylogenetic tree position of the organism(s) in a sample. Those target organisms represented in the original sequence database and sufficient characteristic sequences can identify to the species or subspecies level. Oligonucleotide arrays of many probes are especially preferred. A hybridization signal can comprise fluorescence, chemiluminescence, or isotopic labeling, etc.; or sequences in a sample can be detected by direct means, e.g. mass spectrometry. The method's characteristic sequences can also be used to design specific PCR primers. The method uniquely identifies the phylogenetic affinity of an unknown organism without requiring prior knowledge of what is present in the sample. Even if the organism has not been previously encountered, the method still provides useful information about which phylogenetic tree bifurcation nodes encompass the organism.

Fox, George E.↗

The complete Einstein Observatory X-ray survey of the Orion Nebula region.

We have analyzed archival Einstein Observatory images of a roughly 4.5 square degree region centered on the Orion Nebula. In all, 245 distinct X-ray sources have been detected in six High Resolution Imager (HRI) and 17 Imaging Proportional Counter (IPC) observations. An optical database of over 2700 stars has been assembled to search for candidate counterparts to the X-ray sources. Roughly half the X-ray sources are identified with a single Orion Nebula cluster member. The 10 main-sequence O6-B5 cluster stars detected in Orion have X-ray activity levels comparable to field O and B stars. X-ray emission has also been detected in the direction of four main-sequence late-B and early-A type stars. Since the mechanisms producing X-rays in late-type coronae and early-type winds cannot operate in the late-B and early-A type atmospheres, we argue that the observed X-rays, with L(sub X) approximately = 3 x 10(exp 30) ergs/s, are probably produced in the coronae of unseen late-type binary companions. Over 100 X-ray sources have been associated with late-type pre-main sequence stars. The upper envelope of X-ray activity rises sharply from mid-F to late-G, with L(sub x)/L(sub bol) in the range 10(exp -4) to 2 x 10(exp -3) for stars later than approximately G7. We have looked for variability of the late-type cluster members on timescales of a day to a year and find that 1/4 of the stars show significantly variable X-ray emission. A handful of the late-type stars have published rotational periods and spectroscopic rotational velocities; however, we see no correlation between X-ray activity and rotation. Thus, for this sample of pre-main-sequence stars, the large dispersion in X-ray activity does not appear to be caused by the dispersion in rotation, in contrast with results obtained for low-mass main-sequence stars in the Pleiades and pre-main-sequence stars in Taurus-Auriga.

Gagne, Marc↗

A NASTRAN DMAP alter for linear buckling analysis under dynamic loading

A unique modification to the NASTRAN solution sequence for transient analysis with direct time integration (COSMIC NASTRAN rigid format 9) was developed and incorporated into a DMAP alter. This DMAP alter calculates the buckling stability of a dynamically loaded structure, and is used to predict the onset of structural buckling under stress wave loading conditions. The modified solution sequence incorporates the linear buckling analysis capability (rigid format 5) of NASTRAN into the existing Transient solution rigid format in such a way as to provide a time dependent eigensolution which is used to assess the buckling stability of the structure as it responds to the impulsive load. As a demonstration of the validity of this modified solution procedure, the dynamic buckling of a prismatic bar subjected to an impulsive longitudinal compression is analyzed and compared to the known theoretical solution. In addition, a dynamic buckling analysis is performed for the analytically less tractable problem of the localized dynamic buckling of an initially flawed composite laminate under transverse impact loading. The addition of this DMAP alter to the transient solution sequence in NASTRAN facilitates the prediction of both time and mode of buckling.

Aiello, Robert A.↗

A NASTRAN DMAP alter for linear buckling analysis under dynamic loading

A modification to the NASTRAN solution sequence for transient analysis with direct time integration (COSMIC NASTRAN rigid format 9) was developed and incorporated into a DMAP alter. This DMAP alter calculates the buckling stability of a dynamically loaded structure, and is used to predict the onset of structural buckling under stress-wave loading conditions. The modified solution sequence incorporates the linear buckling analysis capability (rigid format 5) of NASTRAN into the existing Transient solution rigid format in such a way as to provide a time dependent eigensolution which is used to assess the buckling stability of the structure as it responds to the impulsive load. As a demonstration of the validity of this modified solution procedure, the dynamic buckling of a prismatic bar subjected to an impulsive longitudinal compression is analyzed and compared to the known theoretical solution. In addition, a dynamic buckling analysis is performed for the analytically less tractable problem of the localized dynamic buckling of an initially flawed composite laminate under transverse impact loading. The addition of this DMAP alter to the transient solution sequence in NASTRAN facilitates the computational prediction of both the time at which the onset of dynamic buckling occurs in an impulsively loaded structure, and the dynamic buckling mode shapes of that structure.

Aiello, Robert A.↗

Isolation of novel ribozymes that ligate AMP-activated RNA substrates

BACKGROUND: The protein enzymes RNA ligase and DNA ligase catalyze the ligation of nucleic acids via an adenosine-5'-5'-pyrophosphate 'capped' RNA or DNA intermediate. The activation of nucleic acid substrates by adenosine 5'-monophosphate (AMP) may be a vestige of 'RNA world' catalysis. AMP-activated ligation seems ideally suited for catalysis by ribozymes (RNA enzymes), because an RNA motif capable of tightly and specifically binding AMP has previously been isolated. RESULTS: We used in vitro selection and directed evolution to explore the ability of ribozymes to catalyze the template-directed ligation of AMP-activated RNAs. We subjected a pool of 10(15) RNA molecules, each consisting of long random sequences flanking a mutagenized adenosine triphosphate (ATP) aptamer, to ten rounds of in vitro selection, including three rounds involving mutagenic polymerase chain reaction. Selection was for the ligation of an oligonucleotide to the 5'-capped active pool RNA species. Many different ligase ribozymes were isolated; these ribozymes had rates of reaction up to 0.4 ligations per hour, corresponding to rate accelerations of approximately 5 x10(5) over the templated, but otherwise uncatalyzed, background reaction rate. Three characterized ribozymes catalyzed the formation of 3'-5'-phosphodiester bonds and were highly specific for activation by AMP at the ligation site. CONCLUSIONS: The existence of a new class of ligase ribozymes is consistent with the hypothesis that the unusual mechanism of the biological ligases resulted from a conservation of mechanism during an evolutionary replacement of a primordial ribozyme ligase by a more modern protein enzyme. The newly isolated ligase ribozymes may also provide a starting point for the isolation of ribozymes that catalyze the polymerization of AMP-activated oligonucleotides or mononucleotides, which might have been the prebiotic analogs of nucleoside triphosphates.

NASA Discipline Exobiology↗

Predominance of two-armed spirals

A physical process accounts for the origin and widespread occurrence of two-armed spiral patterns with a high degree of symmetry extending from the nucleus to the outermost part of a galaxy. Self-gravitating systems, starting from a wide variety of initial conditions, settle down into twofold symmetries through a sequence of forms that were identified in numerical experiments. Twofold symmetries rarely develop directly. Instead, the systems pass through a sequence of more complicated shapes. The computer experiments and the sequence leading to twofold symmetries are described. At the end of this process, a two-armed spiral density wave with a 'grand design' has been set up in the galaxy, and the stage has been set for some mechanism, such as that of Lin and Shu (1971), to take over to assure a long lifetime for the pattern.

Miller, R. H.↗

A computer aided thermodynamic approach for predicting the formation of Z-DNA in naturally occurring sequences

The ease with which a particular DNA segment adopts the left-handed Z-conformation depends largely on the sequence and on the degree of negative supercoiling to which it is subjected. We describe a computer program (Z-hunt) that is designed to search long sequences of naturally occurring DNA and retrieve those nucleotide combinations of up to 24 bp in length which show a strong propensity for Z-DNA formation. Incorporated into Z-hunt is a statistical mechanical model based on empirically determined energetic parameters for the B to Z transition accumulated to date. The Z-forming potential of a sequence is assessed by ranking its behavior as a function of negative superhelicity relative to the behavior of similar sized randomly generated nucleotide sequences assembled from over 80,000 combinations. The program makes it possible to compare directly the Z-forming potential of sequences with different base compositions and different sequence lengths. Using Z-hunt, we have analyzed the DNA sequences of the bacteriophage phi X174, plasmid pBR322, the animal virus SV40 and the replicative form of the eukaryotic adenovirus-2. The results are compared with those previously obtained by others from experiments designed to locate Z-DNA forming regions in these sequences using probes which show specificity for the left-handed DNA conformation.

Non-NASA Center↗

Hemichordates and the Origin of Chordates

At the start of the period of the NASA grant three years ago, we had no information on the organization and development of the body axis of the hemichordate, Saccoglossus kowalevskii. Now we have substantial findings about the anteroposterior axis and dorsoventral axis, and based on this information, we have new insights about the origin of chordates from ancestral deuterostomes. We found ways to obtain and preserve large numbers of embryos and hatched juveniles. We can now collect about 40,000 embryos in the month of September, the time of S. kowalevskii spawning at Woods Hole. Excellent cDNA libraries were prepared from three developmental stages. From these libraries, we directly isolated about 30 gene ortholog sequences by screening and pcr techniques, all of these sequences of interest in the inquiry about the animal's organization and development. We also performed a mid-sized EST project (60,000 randomly picked clones, many of these arrayed). About half of these have been analyzed so far by blastx and are suitable for direct use of clones. We have obtained about 50 interesting sequences from this set. The rest still await analysis. Thus, at this time we have isolated orthologs of 80 genes that are known to be expressed in chordates in conserved domains and known to have interesting roles in chordate organization and development. The orthology of the S. kowalevskii sequences has been verified by neighbor joining and parsimony methods, with bootstrap estimates of validity. The S. kowalevskii sequences cluster with other deuterostome sequences, namely, other hemichordates, echinoderms, ascidians, amphioxus, or vertebrates, depending on what sequences are available in the database for comparison. We have used these sequences to do high quality in situ hybridization on S. kowalevskii embryos, and the results can be divided into three sections-those concerning the anteroposterior axis of S. kowalevskii in comparison to the same axis of chordates, those concerning the dorsoventral axis of S. kowalevskii in comparison to the same axis of chordates, and those concerning the signals and transcription factors found in the endoderm, of S. kowalevskii compared to the signals and transcription factors in the endo-mesodermal cells of Spemann's organizer of chordates.

Gerhart, John↗

Direct-imaging Discovery and Dynamical Mass of a Substellar Companion Orbiting an Accelerating Hyades Sun-like Star with SCExAO/CHARIS

We present the direct-imaging discovery of a substellar companion in orbit around a Sun-like star member of the Hyades open cluster. So far, no other substellar companions have been unambiguously confirmed via direct imaging around main-sequence stars in Hyades. The star HIP 21152 is an accelerating star as identified by the astrometry from the Gaia and Hipparcos satellites. We detected the companion, HIP 21152 B, in multiple epochs using the high-contrast imaging from SCExAO/CHARIS and Keck/NIRC2. We also obtained the stellar radialvelocity data from the Okayama 188 cm telescope. The CHARIS spectroscopy reveals that HIP 21152 B’s spectrum is consistent with the L/T transition, best fit by an early T dwarf. Our orbit modeling determines the semimajor axis and the dynamical mass of HIP 21152 B to be 17 +7.2 −3.8 and 27.8 +8.4 −5.4 M Jub , respectively. The mass ratio of HIP 21152 B relative to its host is ≈2%, near the planet/brown dwarf boundary suggested by recent surveys. Mass estimates inferred from luminosity-evolution models are slightly higher (33–42 MJup). With a dynamical mass and a well-constrained age due to the system’s Hyades membership, HIP 21152 B will become a critical benchmark in understanding the formation, evolution, and atmosphere of a substellar object as a function of mass and age. Our discovery is yet another key proof of concept for using precision astrometry to select direct-imaging targets.

Masayuki Kuzuhara↗

The Utilization Profiles of the CCSDS Unified Space Link Protocol (USLP)

The purpose of this paper is to identify the utilization profiles for interfacing the Data Protocol Sublayer using the Unified Space Link Protocols (USLP) (reference 1) with the space link coding procedures as specified in the CCSDS Coding & Synchronization Blue Books (references 2 through 5), used in both telecommand and telemetry applications. This paper describes how the USLP Protocol utilizes the coding and synchronization sublayer to support: a. Direct to Earth (DTE) telemetry links for engineering and science data b. Direct to Earth (DTE) telemetry links for very high rate science data c. Direct from Earth (DFE) command, sequencing and flight software loads d. Space to Space Links (Proximity) utilized by orbiters for data exchange to/from surface bound assets. The CCSDS has divided the functions of the Data Link Layer into two sublayers: the Data Link Protocol Sublayer (DLP-SL) and the Coding and Synchronization Sublayer (CS-SL). The Data Link Protocol Sublayer (DLP-SL) interfaces to the users, accepting the data that is to be transported, on the sending side of the link, and delivering that data on the receiving end. The Transfer Frame is the data unit that is transferred across the Data Link Protocol Sublayer and the Coding and Synchronization Sublayer boundary. The Coding and Synchronization Sublayer (CS-SL) provides the encoding, randomization, and frame synchronization functions that prepares the USLP Transfer Frame for transport across the space link. The CS-SL is divided into 2 processes: 1) The Frame Interface Processes (FIP) performs the interface functions required to prepare the data for delivery to the Coding/Decoding Process (CDP). This process includes prepending a Frame Start Marker to the provided frame, when management has designated that the frame is not to be aligned to the codeblock or when there is no block code used. 2) The Coding/Decoding Process (CDP) performs the forward error correction processes that are used to optimize the performance of the link and minimize the error rate. The CDP creates the symbol stream that is delivered to the Physical Layer. The transfer of the USLP transfer frames across different types of space links is the focus of this paper. The Protocol Data Unit (PDU) that is passed in both directions between the Data Link Protocol Sublayer (DLP-SL) and Coding and Synchronization Sublayer (CS-SL) is the transfer frame. The USLP frame structure provides flexibility that can be constrained by the functions utilized within the CS-SL that prepare the transfer frame for transit. For example, the USLP transfer frame contains a length field that enables the frame to be of variable length but CS-SL under certain conditions may constrain the frame to be fixed in length. This paper describes 5 operational modes available for use by the Data Link Layer to provide data exchange across the USLP space link. These modes are different because different operational requirements apply to vastly different types of space links and thus the communications implementation requirements differ. The environmental issues include the power or energy available, the distance between the end points of the link, the complexity of the equipment available at those end points, the atmospheric conditions and radiometric frequency selection. The CS-SL utilizes different forward error correcting codes supported by specific operational modes to configure the data for transit. This paper describes all of the operational modes in a series of data models which decompose the functionality between the Data Link Protocol Sublayer and the Coding and Synchronization sublayer. The operational modes described are: 1. Uncoded Mode: has been used for short links that contain significant available power to provide an acceptable frame error rate. The frames in this mode can be variable in length and typically use an error detection algorithm (i.e., CRC) to determine if there are errors in the received frame. 2. Convolutional Only Mode: is currently the prime forward error correction coding used for the proximity links. The frames in this mode can be variable in length and typically use an error detection algorithm (i.e., CRC) to determine if there are errors in the received frame. 3. Variable Length Frame Aligned to Variable Length Codeblock (TC): is used for Direct from Earth links were power levels are high and the simple, least complex code i.e., the BCH code is used. This mode has been in use since the early 1970s. The BCH code is a short code and the decoder is easy to implement. 4. Fixed Length Frame Aligned to Fixed Length Codeblock (AOS/TM): was introduced when the concatenated Convolutional and Reed-Solomon Code was formulated to provide significant reduction in link data error rate and the ability to determine if there was an error in the decoded codeblock. The frame is aligned to the codeblock so that there is a one to one relationship of frame errors to codeblock errors without additional error detection coding being added. This mode requires the protocol frames to be the exact size of the message portion of the codeblock. 5. Frames Unaligned to Fixed Length Codeblocks (Currently used for very high rates and space to space links): This mode is currently used for missions that have a very high data rate that can be controlled adaptively as the environment changes and as the next generation operating mode for the proximity link. This mode from a coded data stream point of view is exactly like that described in 4. above, except that the frame need not be aligned to the codeblock. There is no requirement on frame length when using this mode. Thus when using USLP it can be used to support links that require short or long frames. There is also no mandatory requirement that frames cannot be separated by idle data reducing the tight data rate connection requirements between the data link protocol sublayer and the coding & synchronization sublayer. In conclusion, how these operational modes can be put to use in mission operational scenarios is described for Direct from Earth links (DFE), Direct to Earth links (DTE), and Proximity links.

Greenberg, E.↗

A geometric theory for the QR, LU and power iterations.

Consideration of the task of computing the invariant subspaces of a given matrix. For this purpose the LU, QR, treppen and bi-iterations have been presented, used, and studied more or less independently of the old-fashioned power method. Each of these methods generates implicitly a sequence of subspaces which determines the convergence properties of the method. The iterations differ in the way in which a basis is constructed to represent each subspace. This aspect largely determines the usefulness of the method. It is shown that the first four iterations produce exactly the same sequence of subspaces as do direct and inverse iteration started from appropriate subspaces. Their convergence properties are therefore the same, and a complete geometric convergence theory is presented in terms of the power method. It is shown that Hessenberg matrices are associated with ideal starting spaces.

Parlett, B. N.↗

Biomedical ultrasonoscope

The combination of a "C" mode scan electronics in a portable, battery powered biomedical ultrasonoscope having "A" and "M" mode scan electronics, the latter including a clock generator for generating clock pulses, a cathode ray tube having X, Y and Z axis inputs, a sweep generator connected between the clock generator and the X axis input of the cathode ray tube for generating a cathode ray sweep signal synchronized by the clock pulses, and a receiver adapted to be connected to the Z axis input of the cathode ray tube. The "C" mode scan electronics comprises a plurality of transducer elements arranged in a row and adapted to be positioned on the skin of the patient's body for converting a pulsed electrical signal to a pulsed ultrasonic signal, radiating the ultrasonic signal into the patient's body, picking up the echoes reflected from interfaces in the patient's body and converting the echoes to electrical signals; a plurality of transmitters, each transmitter being coupled to a respective transducer for transmitting a pulsed electrical signal thereto and for transmitting the converted electrical echo signals directly to the receiver, a sequencer connected between the clock generator and the plurality of transmitters and responsive to the clock pulses for firing the transmitters in cyclic order; and a staircase voltage generator connected between the clock generator and the Y axis input of the cathode ray tube for generating a staircase voltage having steps synchronized by the clock pulses.

Lee, R. D.↗

Dynamic stress analysis of smooth and notched fiber composite flexural specimens

A detailed analysis of the dynamic stress field in smooth and notched fiber composite (Charpy-type) specimens is reported in this paper. The analysis is performed with the aid of the direct transient response analysis solution sequence of MSC/NASTRAN. Three unidirectional composites were chosen for the study. They are S-Glass/Epoxy, Kevlar/Epoxy and T-300/Epoxy composite systems. The specimens are subjected to an impact load which is modeled as a triangular impulse with a maximum of 2000 lb and a duration of 1 ms. The results are compared with those of static analysis of the specimens subjected to a peak load of 2000 lb. For the geometry and type of materials studied, the static analysis results gave close conservative estimates for the dynamic stresses. Another interesting inference from the study is that the impact induced effects are felt by S-Glass/Epoxy specimens sooner than Kevlar/Epoxy or T-300/Epoxy specimens.

Murthy, P. L. N.↗

Dynamic stress analysis of smooth and notched fiber composite flexural specimens

A detailed analysis of the dynamic stress field in smooth and notched fiber composite (Charpy-type) specimens is reported in this paper. The analysis is performed with the aid of the direct transient response analysis solution sequence of MSC/NASTRAN. Three unidirectional composites were chosen for the study. They are S-Glass/Epoxy, Kevlar/Epoxy and T-300/Epoxy composite systems. The specimens are subjected to an impact load which is modeled as a triangular impulse with a maximum of 2000 lb and a duration of 1 ms. The results are compared with those of static analysis of the specimens subjected to a peak load of 2000 lb. For the geometry and type of materials studied, the static analysis results gave close conservative estimates for the dynamic stresses. Another interesting inference from the study is that the impact induced effects are felt by S-Glass/Epoxy specimens sooner than Kevlar/Epoxy or T-300/Epoxy specimens.

Murthy, P. L. N.↗

NASTRAN GPWG tables for combined substructures

A method for computing the mass and center of gravity for basic and combined substructures stored in the NASTRAN Substructure Operating File (SOF) is described. The three step method recovers SOF data blocks for the relevant substructure, processes these data blocks using a specially developed FORTRAN routine, and generates the NASTRAN gridpoint weight generator (GPWG) table for the substructure in a PHASE2 SOF execution using a Direct Matrix Abstraction Program (DMAP) sequence. Verification data for the process is also provided.

Allen, Tom↗

Artificial intelligence approach to planning the robotic assembly of large tetrahedral truss structures

An assembly planner for tetrahedral truss structures is presented. To overcome the difficulties due to the large number of parts, the planner exploits the simplicity and uniformity of the shapes of the parts and the regularity of their interconnection. The planning automation is based on the computational formalism known as production system. The global data base consists of a hexagonal grid representation of the truss structure. This representation captures the regularity of tetrahedral truss structures and their multiple hierarchies. It maps into quadratic grids and can be implemented in a computer by using a two-dimensional array data structure. By maintaining the multiple hierarchies explicitly in the model, the choice of a particular hierarchy is only made when needed, thus allowing a more informed decision. Furthermore, testing the preconditions of the production rules is simple because the patterned way in which the struts are interconnected is incorporated into the topology of the hexagonal grid. A directed graph representation of assembly sequences allows the use of both graph search and backtracking control strategies.

Homemdemello, Luiz S.↗

Adaptive Error Estimation in Linearized Ocean General Circulation Models

Data assimilation methods are routinely used in oceanography. The statistics of the model and measurement errors need to be specified a priori. This study addresses the problem of estimating model and measurement error statistics from observations. We start by testing innovation based methods of adaptive error estimation with low-dimensional models in the North Pacific (5-60 deg N, 132-252 deg E) to TOPEX/POSEIDON (TIP) sea level anomaly data, acoustic tomography data from the ATOC project, and the MIT General Circulation Model (GCM). A reduced state linear model that describes large scale internal (baroclinic) error dynamics is used. The methods are shown to be sensitive to the initial guess for the error statistics and the type of observations. A new off-line approach is developed, the covariance matching approach (CMA), where covariance matrices of model-data residuals are "matched" to their theoretical expectations using familiar least squares methods. This method uses observations directly instead of the innovations sequence and is shown to be related to the MT method and the method of Fu et al. (1993). Twin experiments using the same linearized MIT GCM suggest that altimetric data are ill-suited to the estimation of internal GCM errors, but that such estimates can in theory be obtained using acoustic data. The CMA is then applied to T/P sea level anomaly data and a linearization of a global GFDL GCM which uses two vertical modes. We show that the CMA method can be used with a global model and a global data set, and that the estimates of the error statistics are robust. We show that the fraction of the GCM-T/P residual variance explained by the model error is larger than that derived in Fukumori et al.(1999) with the method of Fu et al.(1993). Most of the model error is explained by the barotropic mode. However, we find that impact of the change in the error statistics on the data assimilation estimates is very small. This is explained by the large representation error, i.e. the dominance of the mesoscale eddies in the T/P signal, which are not part of the 21 by 1" GCM. Therefore, the impact of the observations on the assimilation is very small even after the adjustment of the error statistics. This work demonstrates that simult&neous estimation of the model and measurement error statistics for data assimilation with global ocean data sets and linearized GCMs is possible. However, the error covariance estimation problem is in general highly underdetermined, much more so than the state estimation problem. In other words there exist a very large number of statistical models that can be made consistent with the available data. Therefore, methods for obtaining quantitative error estimates, powerful though they may be, cannot replace physical insight. Used in the right context, as a tool for guiding the choice of a small number of model error parameters, covariance matching can be a useful addition to the repertory of tools available to oceanographers.

Chechelnitsky, Michael Y.↗