Search NASA⌕ Search

SEARCH · Search NASA

Results for “NAD(P)H”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

Nitrate transport is independent of NADH and NAD(P)H nitrate reductases in barley seedlings

Barley (Hordeum vulgare L.) has NADH-specific and NAD(P)H-bispecific nitrate reductase isozymes. Four isogenic lines with different nitrate reductase isozyme combinations were used to determine the role of NADH and NAD(P)H nitrate reductases on nitrate transport and assimilation in barley seedlings. Both nitrate reductase isozymes were induced by nitrate and were required for maximum nitrate assimilation in barley seedlings. Genotypes lacking the NADH isozyme (Az12) or the NAD(P)H isozyme (Az70) assimilated 65 or 85%, respectively, as much nitrate as the wild type. Nitrate assimilation by genotype (Az12;Az70) which is deficient in both nitrate reductases, was only 13% of the wild type indicating that the NADH and NAD(P)H nitrate reductase isozymes are responsible for most of the nitrate reduction in barley seedlings. For all genotypes, nitrate assimilation rates in the dark were about 55% of the rates in light. Hypotheses that nitrate reductase has direct or indirect roles in nitrate uptake were not supported by this study. Induction of nitrate transporters and the kinetics of net nitrate uptake were the same for all four genotypes indicating that neither nitrate reductase isozyme has a direct role in nitrate uptake in barley seedlings.

NASA Discipline Number 61-10↗

Oscillatory shear stress stimulates endothelial production of O2- from p47phox-dependent NAD(P)H oxidases, leading to monocyte adhesion

Arterial regions exposed to oscillatory shear (OS) in branched arteries are lesion-prone sites of atherosclerosis, whereas those of laminar shear (LS) are relatively well protected. Here, we examined the hypothesis that OS and LS differentially regulate production of O2- from the endothelial NAD(P)H oxidase, which, in turn, is responsible for their opposite effects on a critical atherogenic event, monocyte adhesion. We used aortic endothelial cells obtained from C57BL/6 (MAE-C57) and p47phox-/- (MAE-p47-/-) mice, which lack a component of NAD(P)H oxidase. O2- production was determined by dihydroethidium staining and an electron spin resonance using an electron spin trap methoxycarbonyl-2,2,5,5-tetramethyl-pyrrolidine. Chronic exposure (18 h) to an arterial level of OS (+/- 5 dynes/cm2) increased O2- (2-fold) and monocyte adhesion (3-fold) in MAE-C57 cells, whereas chronic LS (15 dynes/cm2, 18 h) significantly decreased both monocyte adhesion and O2- compared with static conditions. In contrast, neither LS nor OS were able to induce O2- production and monocyte adhesion to MAE-p47-/-. Treating MAE-C57 with a cell-permeable superoxide dismutase compound, polyethylene glycol-superoxide dismutase, also inhibited OS-induced monocyte adhesion. In addition, over-expressing p47phox in MAE-p47-/- restored OS-induced O2- production and monocyte adhesion. These results suggest that chronic exposure of endothelial cells to OS stimulates O2- and/or its derivatives produced from p47phox-dependent NAD(P)H oxidase, which, in turn, leads to monocyte adhesion, an early and critical atherogenic event.

NASA Program Fundamental Space Biology↗

Role of xanthine oxidoreductase and NAD(P)H oxidase in endothelial superoxide production in response to oscillatory shear stress

Oscillatory shear stress occurs at sites of the circulation that are vulnerable to atherosclerosis. Because oxidative stress contributes to atherosclerosis, we sought to determine whether oscillatory shear stress increases endothelial production of reactive oxygen species and to define the enzymes responsible for this phenomenon. Bovine aortic endothelial cells were exposed to static, laminar (15 dyn/cm2), and oscillatory shear stress (+/-15 dyn/cm2). Oscillatory shear increased superoxide (O2.-) production by more than threefold over static and laminar conditions as detected using electron spin resonance (ESR). This increase in O2*- was inhibited by oxypurinol and culture of endothelial cells with tungsten but not by inhibitors of other enzymatic sources. Oxypurinol also prevented H2O2 production in response to oscillatory shear stress as measured by dichlorofluorescin diacetate and Amplex Red fluorescence. Xanthine-dependent O2*- production was increased in homogenates of endothelial cells exposed to oscillatory shear stress. This was associated with decreased xanthine dehydrogenase (XDH) protein levels and enzymatic activity resulting in an elevated ratio of xanthine oxidase (XO) to XDH. We also studied endothelial cells lacking the p47phox subunit of the NAD(P)H oxidase. These cells exhibited dramatically depressed O2*- production and had minimal XO protein and activity. Transfection of these cells with p47phox restored XO protein levels. Finally, in bovine aortic endothelial cells, prolonged inhibition of the NAD(P)H oxidase with apocynin decreased XO protein levels and prevented endothelial cell stimulation of O2*- production in response to oscillatory shear stress. These data suggest that the NAD(P)H oxidase maintains endothelial cell XO levels and that XO is responsible for increased reactive oxygen species production in response to oscillatory shear stress.

NASA Discipline Cell Biology↗

Formation of Lignans(-)-Secoisolariciresinol and (-)-Matairesinol with Forsythia intermedia Cell-Free Extracts

In vivo labeling experiments of Forsythia intermedia plant tissue with [8-(C-14)]- and [9,9-(2)H2,OC(2)H3]coniferyl alcohols revealed that the lignans, (-)-secoisolariciresinol and (-)-matairesinol, were derived from two coniferyl alcohol molecules; no evidence for the formation of the corresponding (+)-enantiomers was found. Administration of (+/-)-[Ar-(H-3)] secoisolariciresinols to excised shoots of F.intermedia resulted in a significant conversion into (-)-matairesinol; again, the (+)-antipode was not detected. Experiments using cell-free extracts of F.intermedia confirmed and extended these findings. In the presence of NAD(P)H and H2O2, the cell-free extracts catalyzed the formation of (-)- secoisolariciresinol, with either [8-(C-14)]- or [9,9-(2)H2,OC(2)H3]coniferyl alcohols as substrates. The (+)- enantiomer was not formed. Finally, when either (-)-[Ar-(H-3)] or (+/-)-[Ar-(H-2)]secoisolariciresinols were used as substrates, in the presence of NAD(P), only (-)- and not (+)-matairesinol formation occurred. The other antipode, (+)-secoisolariciresinol, did not serve as a substrate for the formation of either (+)- or (-)-matairesinol. Thus, in F.intermedia, the formation of the lignan, (-)-secoisolariciresinol, occurs under strict stereochemical control, in a reaction or reactions requiring NAD(P)H and H2O2 as cofactors. This stereoselectivity is retained in the subsequent conversion into (-)-matairesinol, since (+)-secoisolariciresinol is not a substrate. These are the first two enzymes to be discovered in lignan formation.

Umezawa, Toshiaki↗

An Extraordinary Accumulation of (-)-Pinoresinol in Cell-Free Extracts of Forsythia intermedia: Evidence for Enantiospecific Reduction of (+)-Pinoresinol

Stereoselective and enantiospecific transformation mechanisms in lignan biogenesis are only now yielding to scientific inquiry: it has been shown that soluble cell-free preparations from Forsythia intermedia catalysis the formation of the enantiomerically pure lignan, (-)-secoisolariciresinol, when incubated with coniferyl alcohol in the presence of NAD(P)H and H2O2. Surprisingly, (-)-pinoresinol also accumulates in this soluble cell-free assay mixture in greater than 96% enantiomeric excess, even though it is not the naturally occurring antipode present in Forsythia sp. But these soluble cell-free preparations do not engender stereoselective coupling; instead, racemic pinoresinols are first formed, catalysed by an H2O2-dependent peroxidase reaction. An enantiospecific NAD(P)H reductase then converts (+)- pinoresinol, and not the (-)-antipode, into (-)-secoisolariciresinol. Stereoselective syntheis of(+)-pinoresinol from E-coniferyl alcohol is, however, catalysed by an insoluble enzyme preparation in F. suspensa, obtained following removal of readily soluble and ionically bound enzymes; no exogenously supplied cofactors were required other than oxygen, although the reaction was stimulated by NAD-malate addition. Thus, the overall biochemical pathway to enantiomerically pure (-)-secoisolariciresinol has been delineated.

Katayama, Takeshi↗

Stereoselectivity in Polyphenol Biosynthesis

Stereoselectivity plays an important role in the late stages of phenyl-propanoid metabolism, affording lignins, lignans, and neolignans. Stereoselectivity is manifested during monolignol (glucoside) synthesis, e.g., where the geometry (E or Z) of the pendant double bond affects the specificity of UDPG:coniferyl alcohol glucosyltransferases in different species. Such findings are viewed to have important ramifications in monolignol transport and storage processes, with roles for both E- and Z-monolignols and their glucosides in lignin/lignan biosynthesis being envisaged. Stereoselectivity is also of great importance in enantiose-lective enzymatic processes affording optically active lignans. Thus, cell-free extracts from Forsythia species were demonstrated to synthesize the enantiomerically pure lignans, (-)-secoisolariciresinol, and (-)-pinoresinol, when NAD(P)H, H2O2 and E-coniferyl alcohol were added. Progress toward elucidating the enzymatic steps involved in such highly stereoselective processes is discussed. Also described are preliminary studies aimed at developing methodologies to determine the subcellular location of late-stage phenylpropanoid metabolites (e.g., coniferyl alcohol) and key enzymes thereof, in intact tissue or cells. This knowledge is essential if questions regarding lignin and lignan tissue specificity and regulation of these processes are to be deciphered.

Lewis, Norman G.↗

Sera from cancer patients contain two oscillating ECTO-NOX activities with different period lengths

ECTO-NOX protein's are cell surface-associated and growth-related hydroquinone oxidases with both protein disulfide-thiol interchange activity and the capacity to oxidize NAD(P)H. The activities of these ECTO-NOX proteins are not steady state but fluctuate to create a repeating pattern of oscillations. Two forms of ECTO-NOX activities have been distinguished. The constitutive ECTO-NOX (CNOX), is hormone responsive and refractory to quinone-site inhibitors. A tumor-associated NOX (tNOX) is unregulated, refractory to hormones and growth factors and responds to quinone-site inhibitors. CNOX proteins are widely distributed and exhibit oscillations in enzymatic activity with a period length of 24 min. tNOX proteins are cancer specific and exhibit oscillations with a period length of about 22 min. Our findings now demonstrate the presence of the novel oscillating tNOX activity in sera of patients with cancer whereas the constitutive NOX of non-cancer cells is present in sera of both cancer patients and healthy volunteers. We conclude that ECTO-NOX proteins in sera exhibit oscillatory characteristics similar to those of ECTO-NOX forms of the cell surface.

NASA Discipline Cell Biology↗

Trypsin digestion for determining orientation of ATPase in Halobacterium saccharovorum membrane vesicles

Membranes prepared by low pressure disruption of cells exhibited no ATPase activity in the absence of Triton X-100, although 43% of the total menadione reductase activity was detected. Trypsin digestion reduced menadione reductase activity by 45% whereas ATPase activity was not affected. Disruption of the membrane fraction at higher pressure solubilized about 45% of the ATPase activity. The soluble activity was still enhanced by Triton X-100, suggesting that the detergent, besides disrupting membrane vesicles, also activated the ATPase. The discrepancy in localization of menadione reductase and ATPase activities raised questions regarding the reliability of using a single marker enzyme as an indicator of vesicle orientation.

NASA Center ARC↗

Considering Combined or Separated Roughness and Vegetation Effects in Soil Moisture Retrievals

For more than six years, the Soil Moisture and Ocean Salinity (SMOS) mission has provided multi angular and full-polarization brightness temperature (TB) measurements at L-band. Geophysical products such as soil moisture (SM) and vegetation optical depth at nadir (tau(sub nad)) are retrieved by an operational algorithm using TB observations at different angles of incidence and polarizations. However, the quality of the retrievals depends on several surface effects, such as vegetation, soil roughness and texture, etc. In the microwave forward emission model used in the retrievals (L-band Microwave Emission Model, L-MEB),soil roughness is modeled with a semi-empirical equation using four main parameters (Q(sub r), H(sub r), N(sub rp), with p = H or V polarizations). At present, these parameters are calibrated with data provided by airborne studies and in situ measurements made at a local scale that is not necessarily representative of the large SMOS footprints (43 km on average) at global scale. In this study, we evaluate the impact of the calibrated values of N(sub rp) and H(sub r) on the SM and tau(sub nad) retrievals based on SMOS TB measurements (SMOS Level 3 product) over the Soil Climate Analysis Network (SCAN) network located in North America over five years (2011-2015). In this study, Qr was set equal to zero and we assumed that N(sub rH)= N(sub rV). The retrievals were performed by varying N(sub rp) from −1 to 2 by steps of 1 and H(sub r) from 0 to 0.6 by steps of 0.1. At satellite scale, the results show that combining vegetation and roughness effects in a single parameter provides the best results in terms of soil moisture retrievals, as evaluated against the in situ SM data. Even though our retrieval approach was very simplified, as we did not account for pixel heterogeneity, the accuracy we obtained in the SM retrievals was almost systematically better than those of the Level 3 product. Improved results were also obtained in terms of optical depth retrievals. These new results may have key consequences in terms of calibration of roughness effects within the algorithms of the SMOS (ESA) and the SMAP (NASA) space missions.

full-polarization↗

Changes in sulfhydryl groups of honeybee glyceraldehyde phosphate dehydrogenase associated with generation of the intermediate plateau in its saturation kinetics

Honeybee and rabbit muscle GPDH were studied to obtain information at the chemical level regarding anomolous saturation kinetics of the honeybee enzyme. Results demonstrate that the enzyme's sulfhydryl groups are implicated in the process. Measured by DTNB titration, native honeybee GPDH has one less active SH than the native rabbit muscle enzyme and displays changes in overall sulfhydryl reactivity after preincubation with G-3-P or G-3-P plus NAD+. The total DTNB reactive sulfhydryls of rabbit muscle GPDH are not changed by preincubation with NAD+ or G-3-P; honeybee GPDH, under certain conductions of preincubation with these ligands, shows a decrease of two total DTNB reactive SH groups. This difference has been confirmed by an independent experiment in which the two enzymes were carboxymethylated with C-14 bromoacetic acid.

Gelb, W. G.↗

Trajectory Studies of Large HNO3-Containing PSC Particles in the Arctic: Evidence for the Role of NAT

Large (5 to >20 micron diameter) nitric-acid-containing polar stratospheric cloud (PSC) particles were observed in the Arctic stratosphere during the winter of 1999-2000. We use a particle growth and sedimentation model to investigate the environment in which these particles grew and the likely phase of the largest particles. Particle trajectory calculations show that, while simulated nitric acid dihydrate (NAD) particle sizes are significantly smaller than the observed maximum particle sizes, nitric acid trihydrate (NAT) particle trajectories are consistent with the largest observed particle sizes.

McKinney, K. A.↗

Correlation of ERTS MSS data and earth coordinate systems

The author has identified the following significant results. Experience has revealed a problem in the analysis and interpretation of ERTS-1 multispectral scanner (MSS) data. The problem is one of accurately correlating ERTS-1 MSS pixels with analysis areas specified on aerial photographs or topographic maps for training recognition computers and/or evaluating recognition results. It is difficult for an analyst to accurately identify which ERTS-1 pixels on a digital image display belong to specific areas and test plots, especially when they are small. A computer-aided procedure to correlate coordinates from topographic maps and/or aerial photographs with ERTS-1 data coordinates has been developed. In the procedure, a map transformation from earth coordinates to ERTS-1 scan line and point numbers is calculated using selected ground control points nad the method of least squares. The map transformation is then applied to the earth coordinates of selected areas to obtain the corresponding ERTS-1 point and line numbers. An optional provision allows moving the boundaries of the plots inward by variable distances so the selected pixels will not overlap adjacent features.

Malila, W. A.↗

On the decay of outbursts in dwarf novae nad X-ray novae

We perform computations using a time-dependent model for the accretion disk limit-cycle mechanism to examine the decay of the optical light following the peak of a dwarf nova outburst. We present the results of a parameter study of the physical input variables which affect the decay rate. In the model, the decay is brought about by a cooling transition front which begins at large radii in the disk and moves inward. The nature of the decay is strongly influenced by the radial dependence of the accretion disk viscosity parameter alpha. To obtain exponential decays for typical dwarf nova parameters, we require alpha proportional to r(exp epsilon(sub 0)), where epsilon(sub 0) approximately = 0.3-0.4. The exact value of epsilon(sub 0) which produces exponential decays depends on factors such as the mass of the accreting star and the inner radius of the accretion disk. Therefore, the observed ubiquity of exponential decays in two different types of systems (dwarf novae and X-ray novae) leads us to believe that alpha is an unnatural scaling for the viscosity. The physics of the cooling transition front must be self-regulating in that the timescale (-parital derivative of lnSigma(r)/partial derivative +)(exp -1) (where Sigma is the surface density) for mass extraction across the front remains constant. This may be consistent with a scaling alpha proportional to (h/r)(exp n), where h is the local disk semi-thickness and n approximately 1-2. As regards the speed of the cooling front, we find v(sub F)(r) proportional to r(exp p), where p approximately 3 at large radii, with an abrupt transition to p approximately 0 at some smaller radius. The r(exp 3) dependence is much steeper than has been found by previous workers and appears to result from the strong variation of specific heat within the cooling front when the front resides at a large radius in the disk. The outflow of disk material across the cooling front causes a significant departure of dln T(sub dff0/dln r from the standard value of -0.75 (expected from steady state accretion) within about 0.2 dex in radius of the break associated with the cooling front -- T(sub eff) aproximately 10(exp 3.9) K (r/10(exp 10 cm)) (exp -0.1). These effects should be observable with eclipse mapping. Finally, it appears that the relatively slow decay rate for the optical flux in the 1975 outburst of A0620-00 can be accounted for if the primary is a approximately 10 Solar mass black hole.

Cannizzo, John K.↗

Galileo photometry of Apollo landing sites

As of December 1992, the Galileo spacecraft performed its second and final flyby (EM2), of the Earth-Moon system, during which it acquired Solid State Imaging (SSI) camera images of the lunar surface suitable for photometric analysis using Hapke's, photometric model. These images, together with those from the first flyby (EM1) in December 1989, provide observations of all of the Apollo landing sites over a wide range of photometric geometries and at eight broadband filter wavelengths ranging from 0.41 micron to 0.99 micron. We have completed a preliminary photometric analysis of Apollo landing sites visible in EM1 images and developed a new strategy for a more complete analysis of the combined EM1 and EM2 data sets in conjunction with telescopic observations and spectrogoniometric measurements of returned lunar samples. No existing single data set, whether from spacecraft flyby, telescopic observation, or laboratory analysis of returned samples, describes completely the light scattering behavior of a particular location on the Moon at all angles of incidence (i), emission (e), and phase angles (a). Earthbased telescopic observations of particular lunar sites provide good coverage of incidence nad phase angles, but their range in emission angle is limited to only a few degrees because of the Moon's synchronous rotation. Spacecraft flyby observations from Galileo are now available for specific lunar features at many photometric geometries unobtainable from Earth; however, this data set lacks coverage at very small phase angles (a less than 13 deg) important for distinguishing the well-known 'opposition effect'. Spectrogoniometric measurements from returned lunar samples can provide photometric coverage at almost any geometry; however, mechanical properties of prepared particulate laboratory samples, such as particle compaction and macroscopic roughness, likely differ from those on the lunar surface. In this study, we have developed methods for the simultaneous analysis of all three types of data: we combine Galileo and telescopic observations to obtain the most complete coverage with photometric geometry, and use spectrogoniometric observations of lunar soils to help distinguish the photometric effects of macroscopic roughness from those caused by particle phase function behavior (i.e., the directional scattering properties of regolith particles).

Helfenstein, P.↗

Correlation between calmodulin activity and gravitropic sensitivity in primary roots of maize

Recent evidence indicates a role for calcium and calmodulin in the gravitropic response of primary roots of maize (Zea mays, L.). We examined this possibility by testing the relationship between calmodulin activity and gravitropic sensitivity in roots of the maize cultivars Merit and B73 x Missouri 17. Roots of the Merit cultivar require light to the gravitropically competent. The gravitropic response of the Missouri cultivar is independent of light. The occurrence of calmodulin in primary roots of these maize cultivars was tested by affinity gel chromatography followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with bovine brain calmodulin as standard. The distribution of calmodulin activity was measured using both the phosphodiesterase and NAD kinase assays for calmodulin. These assays were performed on whole tissue segments, crude extracts, and purified extracts. In light-grown seedlings of the Merit cultivar or in either dark- or light-grown seedlings of the Missouri cultivar, calmodulin activity per millimeter of root tissue was about 4-fold higher in the apical millimeter than in the subtending 3 millimeters. Calmodulin activity was very low in the apical millimeter of roots of dark-grown (gravitropically nonresponsive) seedlings of the Merit cultivar. Upon illumination, the calmodulin activity in the apical millimeter increased to a level comparable to that of light-grown seedlings and the roots became gravitropically competent. The time course of the development of gravitropic sensitivity following illumination paralleled the time course of the increase in calmodulin activity in the apical millimeter of the root. The results are consistent with the suggestion that calmodulin plays an important role in the gravitropic response of roots.

NASA Program Space Biology↗