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At least 55 records · Page 3
All sourcesNASA NTRSDOE OSTI
NASA NTRS2001

Deuterostome evolution: early development in the enteropneust hemichordate, Ptychodera flava

Molecular and morphological comparisons indicate that the Echinodermata and Hemichordata represent closely related sister-phyla within the Deuterostomia. Much less is known about the development of the hemichordates compared to other deuterostomes. For the first time, cell lineage analyses have been carried out for an indirect-developing representative of the enteropneust hemichordates, Ptychodera flava. Single blastomeres were iontophoretically labeled with Dil at the 2- through 16-cell stages, and their fates followed through development to the tornaria larval stage. The early cleavage pattern of P. flava is similar to that of the direct-developing hemichordate, Saccoglossus kowalevskii, as well as that displayed by indirect-developing echinoids. The 16-celled embryo contains eight animal "mesomeres," four slightly larger "macromeres," and four somewhat smaller vegetal "micromeres." The first cleavage plane was not found to bear one specific relationship relative to the larval dorsoventral axis. Although individual blastomeres generate discrete clones of cells, the appearance and exact locations of these clones are variable with respect to the embryonic dorsoventral and bilateral axes. The eight animal mesomeres generate anterior (animal) ectoderm of the larva, which includes the apical organ; however, contributions to the apical organ were found to be variable as only a subset of the animal blastomeres end up contributing to its formation and this varies from embryo to embryo. The macromeres generate posterior larval ectoderm, and the vegetal micromeres form all the internal, endomesodermal tissues. These blastomere contributions are similar to those found during development of the only other hemichordate studied, the direct-developing enteropneust, S. kowalevskii. Finally, isolated blastomeres prepared at either the two- or the four-cell stage are capable of forming normal-appearing, miniature tornaria larvae. These findings indicate that the fates of these cells and embryonic dorsoventral axial properties are not committed at these early stages of development. Comparisons with the developmental programs of other deuterostome phyla allow one to speculate on the conservation of some key developmental events/mechanisms and propose basal character states shared by the ancestor of echinoderms and hemichordates.

NASA Discipline Evolutionary Biology↗
NASA NTRS1969

Basal-plane metallography of deformed pyrolytic carbon

Cleavage technique is recommended over the normal polishing technique in preparing pyrolytic carbon for metallographic examination of basal-plane surfaces. After careful removal of torn basal-plane fragments and other cleavage debris with cellulose tape, the true structure is clearly revealed.

Adkins, J. M.↗
NASA NTRS1974

Morphology and surface mapping

Of the many boulders photographed at the Apollo 17 site, boulder 1 from Station 2 is unique in having a strongly developed foliation. Resistant layers form four steeply inclined ridges separated by joint planes or by deeply eroded beds of softer materials. A prominent cleavage, or set of cross joints, is oriented almost normal to the foliation. The cleavage is expressed as subparallel cracks, some of which are open fissures. The entire surface of the boulder is rough and studded by dark colored knobs ranging in diameter from 1 to 15 cm. It is a polymict breccia containing at least one type of rock that has not been recognized in any other lunar sample, and it records an unusual minor element distribution and magnetic history.

Marvin, U. B.↗
NASA NTRS1974

Cracking and reformation of saturated hydrocarbons by ultrasound in the presence of water

The exposure of saturated hydrocarbons to ultrasound (800 kHz, 6 W/sq cm) in the presence of water results in: (1) cleavage of the carbon chain, producing saturated and unsaturated hydrocarbons with a lower number of carbons than the initial hydrocarbon (cracking); and (2) recombination after cleavage, producing saturated and unsaturated hydrocarbons with a higher number of carbons than the initial hydrocarbon (reformation). The addition of argon facilitates these phenomena. The effects are attributed to a homolytic (radical) mechanism occurring within the cavitation bubbles under the effects of microsparks.

Prudhomme, M. R. O.↗
NASA NTRS1973

Fracture in hexagonal closed packed metals, zinc and beryllium

It is shown that fracture in zinc and beryllium is nucleation controlled and is independent of the nature of the barrier from which fracture nucleates. The double cantilever cleavage technique was used to determine the energy required to propagate a crack on the basal plane (0001) in single crystals. Tensile fracture data from single and asymmetric bicrystals were used to calculate the energy needed to initiate a cleavage crack on the (0001) plane.

Kamdar, M. H.↗
NASA NTRS1983

Surfaces

Techniques for the characterization of surface cleanliness and roughness for predicting the quality of an adhesive bond are outlined. Generally, smooth surfaces are only available from cleavage of crystalline materials along a natural cleavage plane. Films must be deposited on metal surfaces to achieve the same smoothness. Once the surfaces are clean, however, reaction with the ambient atmosphere becomes likely through diffusive and absorption processes, producing asperities. Electron diffraction, Auger electron, and X ray emission spectroscopy are used to characterize surface condition. Once the surface is observed to be clean, the application of an adhesive will usually prohibit separation along the adhesive; separation is then confined to the weaker of the two materials. Finally, the use of polytetrafluorothylene adhesive to test the adhesion between polymers and metal surfaces is described.

Buckley, D. H.↗
NASA NTRS1984

Nucleic acids, proteins, and chirality

The present investigation is concerned with experimental results related, in one case, to the chirality of nucleotides, and, in another case, to the possibility of a link between the chirality of nucleic acids, and that of peptides. It has been found that aminoacylation of the 'internal' hydroxyl group of a dinucleoside monophosphate can occur stereoselectively. However, this reaction has not yet been made a part of a working peptide synthesis scheme. The formation and cleavage of oligonucleotides is considered. In the event of the formation of a helical complex between the oligonucleotide and the polymer, 1-prime,5-prime-bonds in the oligomer are found to become more resistant towards cleavage. The conditions required for peptide bond formation are examined, taking into account the known structures of RNA and possible mechanisms for prebiotic peptide bond formation. The possibility is considered that the 2-prime,5-prime-internucleotide linkage could have played an important part in the early days of biological peptide synthesis.

Usher, D. A.↗
NASA NTRS1986

Evidence for a complex archean deformational history; southwestern Michipicoten Greenstone Belt, Ontario

The Michipicoten Greenstone Belt extends for about 150 km ENE from the northeastern angle of Lake Superior. In common with many other Archean greenstone belts, it is characterized by generally steep bedding dips and a distribution of major lithologic types suggesting a crudely synclinal structure for the belt as a whole. Detailed mapping and determination of structural sequence demonstrates that the structure is much more complex. The Archean history of the belt includes formation of at least three regionally significant cleavages, kilometer-scale overturning, extensive shearing, and diabase intrusion. Most well defined, mappable 'packages' of sedimentary rocks appear to be bounded by faults. These faults were active relatively early in the structural history of the belt, when extensive overturning also occurred. Steepening of dips, NW-SE shortening, development of steep NE cleavage, and pervasive shearing all postdate the early faulting and the regional overturning, obscuring much of the detail needed to define the geometry of the earlier structures. The results obtained so far suggest, however, that the Michipicoten Greenstone Belt underwent an early stage of thrusting and associated isoclinal folding, probably in a convergent tectonic environment.

Mcgill, George E.↗
NASA NTRS1989

Improving the low temperature ductility of NiAl

As part of a study aimed at developing a ductile NiAl-based alloy, ingots of Ni-Fe-Al alloys were cast and hot extruded to rods. The purpose of the iron additions was two-fold viz; to produce a change in the slip vector from 001 to 111 line and, in one alloy, to add a L1(2)-structured ductile second phase. Extruded Ni-20Al-30Fe was two-phase, containing a pro-eutectic B2 phase in a fine lamellar structure of B2+L1(2) phases. Room temperature tensile testing of both single extruded and double extruded alloys resulted in 8-percent and 22-percent plastic elongation and yield stresses of 850 and 760 MPa, respectively. Fracture in both cases occurred by ductile tearing of the eutectic and transgranular cleavage of the proeutectic phase at 1350 MPa. The ductility in double extruded condition is higher than that reported earlier in rapidly solidified wires by Inoue et al. (1984). By comparison, extruded single-phase B2-structured Ni-30Al-20Fe exhibited a fracture strength of 780 MPa, no plasticity, and a mixture of intergranular fracture and transgranular cleavage. This is contrast to earlier work by Inoue et al. where a yield stress of 400 MPa, 5 percent plastic strain, and a mixture of dimple and intergranular fracture was reported.

Guha, Sumit↗
NASA NTRS1991

The role of the gamma/gamma-prime eutectic and porosity on the tensile behavior of a single-crystal nickel-base superalloy

The microstructure of a single-crystal nickel-base superalloy, PWA 1480, has been varied by heat treatment and hot isostatic pressing in order to study the role of the gamma/gamma-prime eutectic and porosity on subsequent tensile behavior. The level of porosity was found not to affect any of the tensile properties, while the gamma/gamma-prime eutectic strongly influenced ductility. Eliminating the gamma/gamma-prime eutectic increased ductility which was attributed to the cleavage fracture of this constituent. It is proposed that such cleavage of the gamma/gamma-prime eutectic is initiated by the stress created from impinging slip bands, promoting shear localization, and final fracture along 111 slip planes. The precise nature of this fracture process is discussed, with emphasis on the role of the gamma/gamma-prime microstructure. The deformation structure of PWA 1480 was also studied, and while different in some respects from many other single-crystal superalloys, its fracture process appears to be similar.

Walston, W. S.↗
NASA NTRS1992

Understanding the organization of the amphibian egg cytoplasm - Gravitational force as a probe

A combination of hypergravity (centrifugation) and hypogravity (clinostat) studies have been carried out on amphibian (frog, Xenopus) eggs. The results reveal that the twinning caused by centrifugation exhibits substantial spawning to spawning variation. That variation can be attributed to the apparent viscosity of the egg's internal cytoplasm. Simulated hypogravity results in a relocation of the egg's third (horizontal) cleavage furrow, toward the equator. Substantial egg-to-egg variation is also observed in this 'cleavage effect'. For interpreting spaceflight data and for using G-forces as probes for understanding the egg's architecture the egg variation documented herein should be considered.

Neff, Anton W.↗
NASA NTRS1991

The Mechanical Properties of a Ni-30Al-20Fe-0.05Zr Intermetallic Alloy in the Temperature Range 300-1200 K

Tensile tests were conducted on an extruded Ni-30(at. pct)Al-20Fe-0.05Zr intermetallic alloy in the temperature range 300-1200 K, and the microstructures were characterized by optical, scanning, and transmission electron microscopy. The alloy did not exhibit any room temperature ductility and failed by transgranular cleavage at a stress of about 710 MPa. This mode of fracture was observed at and below 873 K with the total ductility being less than 2.5 percent. However, the failure mode changed to dimpled fracture, triple point cracking, and rupture above 873 K. Scanning electron microscopy of the fracture surfaces revealed that transgranular cleavage failure had always originated at preexisting defects. In these cases, the fracture stress exhibited a good correlation with the defect size, in accordance with linear elastic fracture mechanics, where the critical stress intensity factor was estimated to be about 4 MPa sq rt m.

Raj, S. V.↗
NASA NTRS1991

Single-parameter characterization of discrete-dislocation pileup tipfield and its application to physically based micro-mechanics

The equivalence between continuous dislocation pileups and cracks is reviewed. The force on the leading dislocation is defined and a general method of calculation of the force is proposed. The equivalence relations are given. Based on the analysis by Eshelby, Frank, and Nabarro and the numerical calculations by Chou and Li and Armstrong et al, it will be shown that the force, F, on the locked leading dislocation of a discrete pileup is capable of characterizing uniquely the stress, strain, and displacement fields at the tip of the pileup, including the positions of the discrete mobile dislocations behind the leading dislocation. Conversely, the positions of the mobile dislocations can be used to measure F. If the propagation of micro-slips and the initiation of micro-fractures at the tip of a pileup are controlled by resolved shear stress and normal cleavage stress respectively, the resolved shear stress intensity coefficient, ReSIC, and the resolved cleavage stress intensity coefficient, ReCIC, must be constant.

Liu, H. W.↗
NASA NTRS1994

The evolution of energy-transducing systems. Studies with archaebacteria

N-ethylmaleimide (NEM) inhibits the ATPase of H. saccharovorum in a nucleotide protectable manner. The bulk of C-14 NEM is incorporated into subunit one. Cyanogen bromide cleavage of labeled subunit one indicated that NEM bound to a peptide of a Mr of about 8,900. Thus, Cys 262 (H. salinarium numbering) may be the NEM binding site. Cyanogen bromide fragments have been submitted for sequencing. To prove the presence of three Cys residues in subunit one, alkaline cleavage following treatment with NTCB was carried out. Thiol reagents such as p-chloro mercuri phenyl sulfonate also inhibited the ATPase. However, this inhibition was not nucleotide-protectable, suggesting a different location and role for the PCMS-sensitive Cys. The proteolipid which was extracted with chloroform/methanol from the membranes of H. saccharovorum cross-reacted with an antiserum against subunit c (the DCCD-binding protein) of Escherichia coli. Following labeling of membranes from H. saccharovorum with C-14 DCCD under conditions, which inhibited ATP synthesis, the isotope was incorporated into one protein of Mr of about 6,500. Thus, the proteolipid of H. saccharovorum and the DCCD-labeled peptide may be identical. If so, these results suggest that the proteolipid is a component of the membrane sector of an archaeal F-type ATP synthase.

Stan-Lotter, Helga↗
NASA NTRS1995

Proton affinity determinations and proton-bound dimer structure indications in C2 to C15, (alpha),(omega)-alkyldiamines

The 'kinetic method' was used to determine the proton affinity (PA) of a,coalkyldiamines from collision induced dissociation (CID) studies of protonated heterodimers. These PA values were consistently lower than those reported in the proton affinity scale. The apparent discrepancy was rationalized in terms of differences in the conformation of the protonated diamine monomers. The minimum energy species, formed by equilibrium proton transfer processes, have a cyclic conformation and the ion charge is shared by both amino-groups which are bridged by the proton. On the other hand, the species formed through dissociation of protonated dimers have a linear structure and the charge is localized on one of the amino-groups. Thus, the difference in the PA values obtained by both methods is a measure of the additional stability acquired by the protonated diamines through cyclization and charge delocalization. The major collision dissociation pathway of the protonated diamine monomers involved elimination of an ammonia moiety. Other reactions observed included loss of the second amino-group and several other bond cleavages. CID of the protonated dimers involved primarily formation of a protonated monomer through cleavage of the weaker hydrogen bond and subsequently loss of ammonia at higher collision energies. As observed from the CID studies, doubly charged ions were also formed from the diamines under conditions of the electrospray ionization.

Karpas, Z.↗
NASA NTRS1994

Early development of Xenopus embryos is affected by simulated gravity

Early amphibian (Xenopus laevis) development under clinostat-simulated weightlessness and centrifuge-simulated hypergravity was studied. The results revealed significant effects on (i) 'morphological patterning' such as the cleavage furrow pattern in the vegetal hemisphere at the eight-cell stage and the shape of the dorsal lip in early gastrulae and (ii) 'the timing of embryonic events' such as the third cleavage furrow completion and the dorsal lip appearance. Substantial variations in sensitivity to simulated force fields were observed, which should be considered in interpreting spaceflight data.

Yokota, Hiroki↗
NASA NTRS1995

A DNA enzyme with Mg(2+)-Dependent RNA Phosphoesterase Activity

Previously we demonstrated that DNA can act as an enzyme in the Pb(2+)-dependent cleavage of an RNA phosphoester. This is a facile reaction, with an uncatalyzed rate for a typical RNA phosphoester of approx. 10(exp -4)/ min in the presence of 1 mM Pb(OAc)2 at pH 7.0 and 23 C. The Mg(2+) - dependent reaction is more difficult, with an uncatalyzed rate of approx. 10(exp -7)/ min under comparable conditions. Mg(2+) - dependent cleavage has special relevance to biology because it is compatible with intracellular conditions. Using in vitro selection, we sought to develop a family of phosphoester-cleaving DNA enzymes that operate in the presence of various divalent metals, focusing particularly on the Mg(2+) - dependent reaction. Results: We generated a population of greater than 10(exp 13) DNAs containing 40 random nucleotides and carried out repeated rounds of selective amplification, enriching for molecules that cleave a target RNA phosphoester in the presence of 1 mM Mg(2+), Mn(2+), Zn(2+) or Pb(2+). Examination of individual clones from the Mg(2+) lineage after the sixth round revealed a catalytic motif comprised of a three-stem junction.This motif was partially randomized and subjected to seven additional rounds of selective amplification, yielding catalysts with a rate of 0.01/ min. The optimized DNA catalyst was divided into separate substrate and enzyme domains and shown to have a similar level of activity under multiple turnover conditions. Conclusions: We have generated a Mg(2+) - dependent DNA enzyme that cleaves a target RNA phosphoester with a catalytic rate approx. 10(exp 5) - fold greater than that of the uncatalyzed reaction. This activity is compatible with intracellular conditions, raising the possibility that DNA enzymes might be made to operate in vivo.

Breaker, Ronald R.↗
NASA NTRS2000

Over-Expression, Purification and Crystallization of Human Dihydrolipoamide Dehydrogenase

Dehydrolipoamide dehydrogenase (E3; dihydrolipoan-tide:NAD+ oxidoreductase, EC 1.8.1.4) is a common catalytic component found in pyruvate dehydrogenase complex, alpha-ketoglutarate dehydrogenase complex, and branched-chain cc-keto acid dehydrogenase complex. E3 is also a component (referred to as L protein) of the glycine cleavage system in bacterial metabolism (2). Active E3 forms a homodimer with four distinctive subdomain structures (FAD binding, NAD+ binding, central and interface domains) with non-covalently but tightly bound FAD in the holoenzyme. Deduced amino acids from cloned full-length human E3 gene showed a total of 509 amino acids with a leader sequence (N-terminal 35 amino acids) that is excised (mature form) during transportation of expressed E3 into mitochondria membrane. So far, three-dimensional structure of human E3 has not been reported. Our effort to achieve the elucidation of the X-ray crystal structure of human E3 will be presented. Recombinant pPROEX-1 expression vector (from GIBCO BRL Life Technologies) having the human E3 gene without leader sequence was constructed by Polymerase Chain Reaction (PCR) and subsequent ligation, and cloned in E.coli XL1-Blue by transformation. Since pPROEX-1 vector has an internal His-tag (six histidine peptide) located at the upstream region of a multicloning site, one-step affinity purification of E3 using nickelnitriloacetic acid (Ni-NTA) agarose resin, which has a strong affinity to His-tag, was feasible. Also a seven-amino-acid spacer peptide and a recombinant tobacco etch virus protease recognition site (seven amino acids peptide) found between His-tag and first amino acid of expressed E3 facilitated the cleavage of His-tag from E3 after the affinity purification. By IPTG induction, ca. 15 mg of human E3 (mature form) was obtained from 1L LB culture with overnight incubation at 25C. Over 98% of purity of E3 from one-step Ni-NTA agarose affinity purification was confirmed by SDS-PAGE analysis. For crystallization, E3 samples were prepared with and without His-tag. To minimize the aggregation of E3, apo- and holo- forms of E3s were tested, as well as a mutated E3. Dynamic light scattering measurements revealed that the E3 preparations without His-tag and substrate are highly monodispersive with regard to homodimers. Consequent crystallization trials of this E3 preparation led to single crystals of E3 grown by the vapor diffusion method. Crystals were obtained within a few days from solution containing poly (ethylene glycol) monomethyl ether 5000 as a precipitant. Autoindexing and integration of the X-ray diffraction data showed that E3 crystals belong to an orthorhombic system with unit cell parameters a-- 123. 1, b= 165.3 and c=214.3A. Further optimization of protein preparation and crystallization experiments for the structural determination will be discussed.

Hong, Y. S.↗