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PubMed1996

A potato cDNA encoding a homologue of mammalian multidrug resistant P-glycoprotein.

A homologue of the multidrug resistance (MDR) gene was obtained while screening a potato stolon tip cDNA expression library with 35S-labeled calmodulin. The mammalian MDR gene codes for a membrane-bound P-glycoprotein (170-180 kDa) which imparts multidrug resistance to cancerous cells. The potato cDNA (PMDR1) codes for a polypeptide of 1313 amino acid residues (ca. 144 kDa) and its structural features are very similar to the MDR P-glycoprotein. The N-terminal half of the PMDR1-encoded protein shares striking homology with its C-terminal half, and each half contains a conserved ATP-binding site and six putative transmembrane domains. Southern blot analysis indicated that potato has one or two MDR-like genes. PMDR1 mRNA is constitutively expressed in all organs studied with higher expression in the stem and stolon tip. The PMDR1 expression was highest during tuber initiation and decreased during tuber development.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2003

Flexor bias of joint position in humans during spaceflight.

The ability to estimate ankle and elbow joint position was tested before, during, and after a 17-day spaceflight. Subjects estimated targeted joint angles during isovelocity (IsoV) joint movements with agonist muscle groups either active or relaxed. These movements included elbow extension (EE) and elbow flexion (EF), and plantarflexion (PF) and dorsiflexion (DF) of the ankle. Subjects also estimated these joint positions while moving the dynamometer at their chosen (variable) velocity (VarV) during EE and PF. For IsoV tests, no differences were observed between active and passive movements for either the ankle or elbow. Compared with those of pre-flight test days, estimates of targeted elbow joint angles were approximately 5 degrees to 15 degrees more flexed in-flight, and returned toward the pre-flight values during recovery. The spaceflight effects for the ankle were inconsistent and less prevalent than those for the elbow. The VarV PF test condition for the 120 degrees target angle at the ankle exhibited approximately 5 degrees to 7 degrees more DF target angle estimates in-flight compared with those pre- or post-flight. In contrast, during IsoV PF there was a tendency for ankle estimates to be approximately 2 degrees to 3 degrees more PF after 2-3 days exposure to spaceflight. These data indicate that during spaceflight the perception of elbow extension is greater than actuality, and are consistent with the interpretation that microgravity induced a flexor bias in the estimation of the actual elbow joint position. Moreover, these effects in joint proprioception during spaceflight were observed in individual isolated single-joint movements during tasks in which vestibular function in maintaining posture were minimal.

Adult↗

PubMed2000

Complement activation promotes muscle inflammation during modified muscle use.

Modified muscle use can result in muscle inflammation that is triggered by unidentified events. In the present investigation, we tested whether the activation of the complement system is a component of muscle inflammation that results from changes in muscle loading. Modified rat hindlimb muscle loading was achieved by removing weight-bearing from the hindlimbs for 10 days followed by reloading through normal ambulation. Experimental animals were injected with the recombinant, soluble complement receptor sCR1 to inhibit complement activation. Assays for complement C4 or factor B in sera showed that sCR1 produced large reductions in the capacity for activation of the complement system through both the classical and alternative pathways. Analysis of complement C4 concentration in serum in untreated animals showed that the classical pathway was activated during the first 2 hours of reloading. Analysis of factor B concentration in untreated animals showed activation of the alternative pathway at 6 hours of reloading. Administration of sCR1 significantly attenuated the invasion of neutrophils (-49%) and ED1(+) macrophages (-52%) that occurred in nontreated animals after 6 hours of reloading. The presence of sCR1 also reduced significantly the degree of edema by 22% as compared to untreated animals. Together, these data show that increased muscle loading activated the complement system which then briefly contributes to the early recruitment of inflammatory cells during modified muscle loading.

Animals↗

PubMed1992

A phylogenetic analysis of Aquifex pyrophilus.

The 16S rRNA of the bacterion Aquifex pyrophilus, a microaerophilic, oxygen-reducing hyperthermophile, has been sequenced directly from the the PCR amplified gene. Phylogenetic analyses show the Aq. pyrophilus lineage to be probably the deepest (earliest) in the (eu)bacterial tree. The addition of this deep branching to the bacterial tree further supports the argument that the Bacteria are of thermophilic ancestry.

Archaea↗

PubMed2001

Development of a space radiation Monte Carlo computer simulation based on the FLUKA and ROOT codes.

This NASA funded project is proceeding to develop a Monte Carlo-based computer simulation of the radiation environment in space. With actual funding only initially in place at the end of May 2000, the study is still in the early stage of development. The general tasks have been identified and personnel have been selected. The code to be assembled will be based upon two major existing software packages. The radiation transport simulation will be accomplished by updating the FLUKA Monte Carlo program, and the user interface will employ the ROOT software being developed at CERN. The end-product will be a Monte Carlo-based code which will complement the existing analytic codes such as BRYNTRN/HZETRN presently used by NASA to evaluate the effects of radiation shielding in space. The planned code will possess the ability to evaluate the radiation environment for spacecraft and habitats in Earth orbit, in interplanetary space, on the lunar surface, or on a planetary surface such as Mars. Furthermore, it will be useful in the design and analysis of experiments such as ACCESS (Advanced Cosmic-ray Composition Experiment for Space Station), which is an Office of Space Science payload currently under evaluation for deployment on the International Space Station (ISS). FLUKA will be significantly improved and tailored for use in simulating space radiation in four ways. First, the additional physics not presently within the code that is necessary to simulate the problems of interest, namely the heavy ion inelastic processes, will be incorporated. Second, the internal geometry package will be replaced with one that will substantially increase the calculation speed as well as simplify the data input task. Third, default incident flux packages that include all of the different space radiation sources of interest will be included. Finally, the user interface and internal data structure will be melded together with ROOT, the object-oriented data analysis infrastructure system. Beyond the benefits of 'objectivity', ROOT's incorporation will also provide a graphical user interface with powerful tools for input prior to the calculation, as well as for data analysis and visualization of the results.

Computer Simulation↗

PubMed1996

A mutation in protein phosphatase 2A regulatory subunit A affects auxin transport in Arabidopsis.

The phytohormone auxin controls processes such as cell elongation, root hair development and root branching. Tropisms, growth curvatures triggered by gravity, light and touch, are also auxin-mediated responses. Auxin is synthesized in the shoot apex and transported through the stem, but the molecular mechanism of auxin transport is not well understood. Naphthylphthalamic acid (NPA) and other inhibitors of auxin transport block tropic curvature responses and inhibit root and shoot elongation. We have isolated a novel Arabidopsis thaliana mutant designated roots curl in NPA (rcn1). Mutant seedlings exhibit altered responses to NPA in root curling and hypocotyl elongation. Auxin efflux in mutant seedlings displays increased sensitivity to NPA. The rcn1 mutation was transferred-DNA (T-DNA) tagged and sequences flanking the T-DNA insert were cloned. Analysis of the RCN1 cDNA reveals that the T-DNA insertion disrupts a gene for the regulatory A subunit of protein phosphatase 2A (PP2A-A). The RCN1 gene rescues the rcn1 mutant phenotype and also complements the temperature-sensitive phenotype of the Saccharomyces cerevisiae PP2A-A mutation, tpd3-1. These data implicate protein phosphatase 2A in the regulation of auxin transport in Arabidopsis.

Amino Acid Sequence↗

PubMed1989

Molecular modeling of calmodulin: a comparison with crystallographic data.

Two methods of side-chain placement on a modeled protein have been examined. Two molecular models of calmodulin were constructed that differ in the treatment of side chains prior to optimization of the molecule. A virtual bond analysis program developed by Purisima and Scheraga was used to determine the backbone conformation based on 2.2 angstroms resolution C alpha coordinates for the molecules. In the first model, side chains were initially constructed in an extended conformation. In the second model, a conformational grid search technique was employed. Calcium ions were treated explicitly during energy optimization using CHARMM. The models are compared to a recently published refined crystal structure of calmodulin. The results indicate that the initial choices for side-chains, but also significant effects on the main-chain conformation and supersecondary structure. The conformational differences are discussed. Analysis of these and other methods makes possible the formulation of a methodology for more appropriate side-chain placement in modeled proteins.

Amino Acid Sequence↗

PubMed1999

A role for cysteine 3635 of RYR1 in redox modulation and calmodulin binding.

Oxidation of the skeletal muscle Ca(2+) release channel (RYR1) increases its activity, produces intersubunit disulfide bonds, and blocks its interaction with calmodulin. Conversely, bound calmodulin protects RYR1 from the effects of oxidants (Zhang, J.-Z., Wu, Y., Williams, B. Y., Rodney, G., Mandel, F., Strasburg, G. M., and Hamilton, S. L. (1999) Am. J. Physiol. 276, Cell Physiol. C46-C53). In addition, calmodulin protects RYR1 from trypsin cleavage at amino acids 3630 and 3637 (Moore, C. P., Rodney, G., Zhang, J.-Z., Santacruz-Toloza, L., Strasburg, G. M., and Hamilton, S. L. (1999) Biochemistry 38, 8532-8537). The sequence between these two tryptic sites is AVVACFR. Alkylation of RYR1 with N-ethylmaleimide (NEM) blocks both (35)S-apocalmodulin binding and oxidation-induced intersubunit cross-linking. In the current work, we demonstrate that both cysteines needed for the oxidation-induced intersubunit cross-link are protected from alkylation with N-ethylmaleimide by bound calmodulin. We also show, using N-terminal amino acid sequencing together with analysis of the distribution of [(3)H]NEM labeling with each sequencing cycle, that cysteine 3635 of RYR1 is rapidly labeled by NEM and that this labeling is blocked by bound calmodulin. We propose that cysteine 3635 is located at an intersubunit contact site that is close to or within a calmodulin binding site. These findings suggest that calmodulin and oxidation modulate RYR1 activity by regulating intersubunit interactions in a mutually exclusive manner and that these interactions involve cysteine 3635.

Alkylation↗

PubMed1992

Inherent limitations of nondestructive chlorophyll meters: a comparison of two types of meters.

Two types of nondestructive chlorophyll meters were compared with a standard, destructive chlorophyll measurement technique. The nondestructive chlorophyll meters were 1) a custom built, single-wavelength meter, and 2) the recently introduced, dual-wavelengh, chlorophyll meter from Minolta (model SPAD-502). Data from both meters were closely correlated with destructive measurements of chlorophyll (r2 = 0.90 and 0.93; respectively) for leaves with chlorophyll concentrations ranging from 100 to 600 mg m-2, but both meters consistently overestimated chlorophyll outside this range. Although the dual-wavelength meter was slightly more accurate than the single-wavelength meter (higher r2), the light-scattering properties of leaf cells and the nonhomogeneous distribution of chlorophyll in leaves appear to limit the ability of all meters to estimate in vivo chlorophyll concentration.

Chlorophyll↗

PubMed1995

Photodissociation of cyanoacetylene: application to the atmospheric chemistry of Titan.

The quantum yield and reaction threshold for the photochemical dissociation of cyanoacetylene into a hydrogen atom and the cyanoethynyl radical have been determined. The quantum yield at 185 nm is approximately 0.09. The threshold is approximately 240 nm. Combination of this data with literature values shows that production of excited-state cyanoacetylene is the major primary process resulting from irradiation between 185 and 254 nm. Also determined are the relative rate constants for the abstraction of a hydrogen atom from hydrogen, methane, and ethane by the cyanoethynyl radical (k(H2):k(CH4):k(C2H6) = 1:9.3:63). Implications of these results for the proposal that hydrogen abstraction plays an important role in the conversion of methane to ethane and in the protection of unsaturated compounds from photoconsumption in the atmosphere of Titan are discussed.

Acetylene↗

PubMedDate not supplied

Combined dietary and exercise intervention for control of serum cholesterol in the workplace.

PURPOSE: To elucidate a potential combined dietary and exercise intervention affect on cardiovascular risk reduction of the National Aeronautics and Space Administration Headquarters employees. DESIGN: A nonexperimental, longitudinal, clinical-chart review study (1987 to 1996) of an identified intervention group and a reference (not a control) group. SETTING: The study group worked in an office environment and participated in the annual medical examinations. SUBJECTS: An intervention group of 858 people with initially elevated serum cholesterol, and a reference group of 963 people randomly sampled from 10% of the study group. MEASURES: Serum cholesterol data were obtained for both groups, respectively, from pre- and postintervention and annual examinations. The reference group was adjusted by statistical exclusion of potential intervention participants. Regression equations (cholesterol vs. study years) for the unadjusted/adjusted reference groups were tested for statistical significance. INTERVENTION: An 8-week individualized, combined dietary and exercise program was instituted with annual follow-ups and was repeated where warranted. RESULTS: Only the unadjusted (but not the adjusted) reference group with initial mean total serum cholesterol levels above 200 mg/dL shows a significant 9-year decline trend and significant beta coefficient tests. An intervention effect is suggested. Mean high density lipoprotein cholesterol rose slightly in the intervention group but was maintained in the reference group. CONCLUSION: With potential design limitations, the NASA intervention program focusing on a high risk group may be associated to some degree, if not fully, with an overall cardiovascular risk profile improvement.

Cardiovascular Diseases↗

PubMed2003

Short-period circumnutations found in sunflower hypocotyls in satellite orbit. A reappraisal of data from Spacelab-1.

We have further analysed data from an experiment performed in satellite orbit, in Spacelab-1. In micro-gravity the hypocotyls of Helianthus annuus, cv. "Teddy Bear", showed short period circumnutations (periods around 30 minutes) as well as the already reported long period nutations (with an average period of about 115 minutes). We applied various types of signal analysis (Fourier and wavelet analysis) to the data series. The long period circumnutations have a larger amplitude than the short term circumnutations. Both short and long period circumnutations exist in one and the same hypocotyl. (This is in contrast to our ground control experiments, where were found only the long-period nutations.) The period of the nutations changed throughout the experiment. These results are extending the conclusions drawn after the Spacelab experiment (Brown et al. 1990). In particular they emphasize the existence of both short- and long-period circumnutations in micro-gravity.

Fourier Analysis↗

PubMed1991

Pilot personality and crew coordination: implications for training and selection.

The performance of pilots can be construed as a product of skill, attitude, and personality factors. Although a great deal of effort within the aviation community has been focused on ensuring technical expertise, and new efforts highlight attitudes associated with crew coordination, personality factors have been relatively unexplored. Further, it is argued that past failures to find linkages between personality and performance were due to a combination of inadequate statistical modeling, premature performance evaluation, and/or the reliance on data gathered in contrived as opposed to realistic situations. The goal of the research presented in this article is to isolate subgroups of pilots along performance-related personality dimensions and to document limits on the impact of crew coordination training between the groups. Two samples of military pilots were surveyed in the context of training in crew coordination. Three different profiles were identified through cluster analysis of personality scales. These clusters replicated across samples and predicted attitude change following training in crew coordination.

Attitude↗

PubMed1991

Phylogenetic diversity in the genus Bacillus as seen by 16S rRNA sequencing studies.

Comparative sequence analysis of 16S ribosomal (r)RNAs or DNAs of Bacillus alvei, B. laterosporus, B. macerans, B. macquariensis, B. polymyxa and B. stearothermophilus revealed the phylogenetic diversity of the genus Bacillus. Based on the presently available data set of 16S rRNA sequences from bacilli and relatives at least four major "Bacillus clusters" can be defined: a "Bacillus subtilis cluster" including B. stearothermophilus, a "B. brevis cluster" including B. laterosporus, a "B. alvei cluster" including B. macerans, B. maquariensis and B. polymyxa and a "B. cycloheptanicus branch".

Bacillus↗

PubMed1991

Isotopic analysis of cometary organic matter.

Carbon isotope ratios have been measured for CN in the coma of comet Halley and for several CHON particles emitted by Halley. Of these, only the CHON-particle data may be reasonably related to organic matter in the cometary nucleus, but the true range of 13C/12C values in those particles is quite uncertain. The D/H ratio in H2O in the Halley coma resembles that in Titan/Uranus. The next decade should substantially improve our understanding of the distribution of C, H, N, and O isotopes in cometary organics. The isotopic composition of meteoritic organic matter is better understood and can serve as a useful analog for the cometary case.

Carbon↗

PubMed1994

Long-term modulation of Galactic Cosmic Radiation and its model for space exploration.

As the human exploration of space has received new attention in the United States, studies find that exposure to space radiation could adversely impact the mission design. Galactic Cosmic Radiation (GCR), with its very wide range of charges and energies, is particularly important for a mission to Mars, because it imposes a stiff mass penalty for spacecraft shielding. Dose equivalent versus shielding thickness calculations, show a rapid initial drop in exposure with thickness, but an asymptotic behavior at a higher shielding thickness. Uncertainties in the radiobiology are largely unknown. For a fixed radiation risk, this leads to large uncertain ties in shielding thickness for small uncertainties in estimated dose. In this paper we investigate the application of steady-state, spherically-symmetric diffusion-convection theory of solar modulation to individual measurements of differential energy spectra from 1954 to 1989 in order to estimate the diffusion coefficient, kappa (r,t), as a function of time. We have correlated the diffusion coefficient to the Climax neutron monitor rates and show that, if the diffusion coefficient can be separated into independent functions of space and time: kappa (-r,t)=K(t)kappa 0 beta P kappa 1(r), where beta is the particle velocity and P the rigidity, then (i) The time dependent quantity 1/K(t), which is proportional to the deceleration potential, phi(r,t), is linearly related to the Climax neutron monitor counting rate. (ii) The coefficients obtained from hydrogen or helium intensity measurements are the same. (iii) There are different correlation functions for odd and even solar cycles. (iv) The correlation function for the Climax neutron monitor counting rate for given time, t, can be used to estimate mean deceleration parameter phi(t) to within +/- 15% with 90% confidence. We have shown that kappa(r,t) determined from hydrogen and/or helium data, can be used to fit the oxygen and iron differential energy spectra with a root mean square error of about +/- 10%, and essentially independent of the particle charge or energy. We have also examined the ion chamber and 14C measurements which allow the analysis to be extended from the year 1906 to 1990. Using this model we have defined reference GCR spectra at solar minimum and solar maximum. These can be used for space exploration studies and provide a quantitative estimate of the error in dose due to changes in GCR intensities.

Carbon Radioisotopes↗

PubMed2000

Vascular mechanics of the coronary artery.

UNLABELLED: This paper describes our research into the vascular mechanics of the coronary artery and plaque. The three sections describe the determination of arterial mechanical properties using intravascular ultrasound (IVUS), a constitutive relation for the arterial wall, and finite element method (FEM) models of the arterial wall and atheroma. METHODS: Inflation testing of porcine left anterior descending coronary arteries was conducted. The changes in the vessel geometry were monitored using IVUS, and intracoronary pressure was recorded using a pressure transducer. The creep and quasistatic stress/strain responses were determined. A Standard Linear Solid (SLS) was modified to reproduce the non-linear elastic behavior of the arterial wall. This Standard Non-linear Solid (SNS) was implemented into an axisymetric thick-walled cylinder numerical model. Finite element analysis models were created for five age groups and four levels of stenosis using the Pathobiological Determinants of Atherosclerosis Youth (PDAY) database. RESULTS: The arteries exhibited non-linear elastic behavior. The total tissue creep strain was epsilon creep = 0.082 +/- 0.018 mm/mm. The numerical model could reproduce both the non-linearity of the porcine data and time dependent behavior of the arterial wall found in the literature with a correlation coefficient of 0.985. Increasing age had a strong positive correlation with the shoulder stress level, (r = 0.95). The 30% stenosis had the highest shoulder stress due to the combination of a fully formed lipid pool and a thin cap. CONCLUSIONS: Studying the solid mechanics of the arterial wall and the atheroma provide important insights into the mechanisms involved in plaque rupture.

Animals↗

PubMed1994

Radiation effects on late cytopathological parameters in the murine lens relative to particle fluence.

Lenses of mice irradiated with 250 MeV protons, 670 MeV/amu 20Ne, 600 MeV/amu 56Fe, 600 MeV/amu 93Nb and 593 MeV/amu 139La ions were evaluated by analyzing cytopathological indicators which have been implicated in the cataractogenic process. The LETs ranged from 0.40 keV/micrometer to 953 keV/micrometer and fluences from 1.31 10(3)/mm2 to 4.99 x 10(7)/mm2. 60Co gamma-rays were used as the reference radiation. The doses ranged from 10 to 40 cGy. The lenses were assessed 64 weeks post irradiation in order to observe the late effects of LET and dose on the target cell population of the lens epithelium. Our study shows that growth dependent pathological changes occur at the cellular level as a function of dose and LET. The shapes of the RBE-LET and RBE-dose curves are consistent with previous work on eye and other biological systems done in both our laboratory and others. The RBEmax's were estimated, for the most radiation cataract related cytological changes, MN frequency and MR disorganization, by calculating the ratio of the initial slopes of dose effect curve for various heavy ions to that of 60Co gamma-ray. For each ion studied, the RBEmax derived from micronucleus (MN) frequency is similar to that derived from meridional row (MR) disorganization, suggesting that heavy ions are equally efficient at producing each type of damage. Furthermore, on a per particle basis (particle/cell nucleus), both MN frequency and MR disorganization are LET dependent indicating that these classic precataractogenic indicators are multi-gene effects. Poisson probability analysis of the particle number traversing cell nuclei (average area = 24 micrometers2) suggested that single nuclear traversals determine these changes. By virtue of their precataractogenic nature the data on these endpoints intimate that radiation cataract may also be the consequence of single hits. In any case, these observations are consistent with the current theory of the mechanism of radiation cataractogenesis, which proposes that genomic damage to the epithelial cells surviving the exposure is responsible for opacification.

Animals↗