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Effects of space food bar feeding on bone mass and metabolism in normal and unloaded rats.

During spaceflights in the shuttle, rats are provided specific food bars. To determine whether this diet allows normal body and skeletal growth, we used four groups of rats fed either standard pellet food or space food bars during a 2-wk unloading experiment. We recorded food intake, body weight, tibial bone mass, and mineral content by ash analyses, cancellous bone volume, and cell activities by histomorphometry. We found that food intake was not different when comparing the two types of food, but that suspended animals had a lower food intake than normal loaded animals. Body weight and bone mass were found lower in suspended animals than in normal loaded animals. Finally, longitudinal growth rate, cancellous bone volume, and bone formation rate were lower in suspended animals, irrespective of the type of food. These results show that space food bar feeding did not affect normal body and skeletal growth, and that body and bone changes due to unloading were not significantly different in animals fed space food bars and standard food.

Animal Feed↗

Animal models for the study of the effects of spaceflight on the immune system.

Animal models have been used to determine the effects of spaceflight on the immune system. Rats and rhesus monkeys have been the primary animals used for actual space flight studies, but mice have also been utilized for studies in ground-based models. The primary ground based model used has been hindlimb unloading of rodents, which is similar to the chronic bed-rest model for humans. A variety of immune responses have been shown to be modified when animals are hindlimb unloaded. These results parallel those observed when animals are flown in space. In general, immune responses are depressed in animals maintained in the hindlimb unloading model or flown in space. These results raise the possibility that spaceflight could result in decreased resistance to infection in animals.

Animals↗

A codiffusing system as a novel model for capillary wall charge selectivity.

The electrostatic interaction associated with polyion-polyion interaction has been thought to be the basis for the differential transport of charged transport probes across the capillary wall. The charge of the transport probe may interact with the negative charges of capillary wall cell surfaces and the intercellular glycocalyx or extracellular matrix. This study has set out to quantitate the exact nature of the polyion-polyion interaction through the theoretical and experimental study of the partition-diffusion of albumin in solutions of anionic polysaccharides (dextran sulfate or heparin) at concentrations up to 250 meq 1(-1) or at an average intercharge distance of 2.2 nm, as compared with solutions of uncharged dextran at the same volume concentration. The results demonstrate that the albumin partition-diffusion is exactly the same in dextran sulfate, heparin and uncharged dextran matrices of the same polymer volume fraction. These results confirm previous studies from this laboratory that the charge effect through polyion-polyion interaction under physiological conditions is negligible.

Albumins↗

Human respiratory airflow through an artificial nasal model: pressure/flow relationship.

Nasal airflow and differential pressure at peak flow were measured by Rhinorheograph MPR-1100 made by Nihon Kohden Co., Ltd. with human resting respiratory airflow through an artificial nasal model. In the healthy nasal model, it was assumed that nasal resistance was produced mainly in the vicinity of the pyriform aperture. Reduction of air space in the nasal model due to various simulated diseases led to decrease airflow and increased differential pressure with consequent increase of nasal resistance. Severe diffuse swelling of the inferior turbinate was the most important cause of marked elevation of resistance, nasal secretion also was an important factor which increased nasal resistance.

Airway Resistance↗

A review of the effects of microgravity and of hypergravity on aging and longevity.

This article reviews the literature dealing with the effects of acute or chronic exposure to microgravity (spacecrafts) or hypergravity (centrifuge) on longevity and the aging process. Even if space flights are now common, the effects of these two kinds of environment on aging are still poorly documented, particularly in mammals. However, there is a growing interest for the use of the fruit fly Drosophila melanogaster, and this species may be now considered as a model organism in gravitational biology studies dealing with aging.

Aging↗

Source separation in astrophysical maps using independent factor analysis.

A microwave sky map results from a combination of signals from various astrophysical sources, such as cosmic microwave background radiation, synchrotron radiation and galactic dust radiation. To derive information about these sources, one needs to separate them from the measured maps on different frequency channels. Our insufficient knowledge of the weights to be given to the individual signals at different frequencies makes this a difficult task. Recent work on the problem led to only limited success due to ignoring the noise and to the lack of a suitable statistical model for the sources. In this paper, we derive the statistical distribution of some source realizations, and check the appropriateness of a Gaussian mixture model for them. A source separation technique, namely, independent factor analysis, has been suggested recently in the literature for Gaussian mixture sources in the presence of noise. This technique employs a three layered neural network architecture which allows a simple, hierarchical treatment of the problem. We modify the algorithm proposed in the literature to accommodate for space-varying noise and test its performance on simulated astrophysical maps. We also compare the performances of an expectation-maximization and a simulated annealing learning algorithm in estimating the mixture matrix and the source model parameters. The problem with expectation-maximization is that it does not ensure global optimization, and thus the choice of the starting point is a critical task. Indeed, we did not succeed to reach good solutions for random initializations of the algorithm. Conversely, our experiments with simulated annealing yielded initialization-independent results. The mixing matrix and the means and coefficients in the source model were estimated with a good accuracy while some of the variances of the components in the mixture model were not estimated satisfactorily.

Astronomy↗

Speed congenics: accelerated genome recovery using genetic markers.

Genetic markers throughout the genome can be used to speed up 'recovery' of the recipient genome in the backcrossing phase of the construction of a congenic strain. The prediction of the genomic proportion during backcrossing depends on the assumptions regarding the distribution of chromosome segments, the population structure, the marker spacing and the selection strategy. In this study simulation was used to investigate the rate of recovery of the recipient genome for a mouse, Drosophila and Arabidopsis genome. It was shown that an incorrect assumption of a binomial distribution of chromosome segments, and failing to take account of a reduction in variance in genomic proportion due to selection, can lead to a downward bias of up to two generations in the estimation of the number of generations required for the formation of a congenic strain.

Animals↗

Peak separation and sorting by coherent 2D resonance Raman spectroscopy.

The ability to separate and sort peaks is explored using a new coherent two-dimensional form of resonance Raman spectroscopy. This experimental technique distributes normally congested rotational-vibrational peaks along a series of curved lines according to vibrational sequence, rotational quantum number, and selection rule. Each line consists of rotational-vibrational peaks that have the same vibrational sequence and the same value for DeltaJ, distributed in order by rotational quantum number. For diatomic molecules, these lines originate from points where they initially travel in opposite or orthogonal directions in two-dimensional space, which helps facilitate the separation between lines. Simulations and experimental results on C2 in a flame confirm the ability to separate and sort these normally congested rotational-vibrational peaks. This method appears to provide a solution to the long-standing problems of spectral congestion and disorder in gas-phase electronic spectra.

Computer Simulation↗

Structural influence of RNA incorporation in DNA: quantitative nuclear magnetic resonance refinement of d(CG)r(CG)d(CG) and d(CG)r(C)d(TAGCG).

RNA and DNA adopt different types of conformations, i.e., A-type with C3'-endo sugar pucker for RNA and B-type with C2'-endo sugar pucker for DNA, respectively. The structural influence of the incorporation of RNA nucleotides into DNA is less understood. In this paper, we present the three-dimensional structures of two RNA-containing oligonucleotides, d(CG)r(CG)d(CG) and d(CG)r(C)d-(TAGCG), as determined by the NMR refinement procedure, and assess the possible structural perturbation of DNA induced by RNA. With a single RNA insertion into an octamer DNA, its overall conformation remains as the canonical B-DNA, except that the sugar pucker of the rC3 residue is C3'-endo (pseudorotation angle P = 3.6 degrees). In contrast, the hybrid hexamer is neither the pure B-DNA nor the pure A-DNA conformation. Instead, we propose a model in which the DNA parts adopt B conformation, whereas the RNA part adopts A conformation, with the overall conformation closer to A-DNA. To ensure an exhaustive search of the conformational space, the model was subjected to 100-ps simulated annealing with slow cooling or 100-ps molecular dynamics with subsequent quenching. Models obtained at different time points of the trajectories were further subjected to the SPEDREF NOE refinement [Robinson & Wang (1992) Biochemistry 31, 3524] and they appeared to arrive at a convergent model (< 0.5 A RMSD for the central four base pairs). The consensus hexamer structure contains a significant discontinuity at the (rG4)p(dC5) step with a base pair tilt angle of 6.7 degrees and roll angle of 11.5 degrees.(ABSTRACT TRUNCATED AT 250 WORDS)

Cold Temperature↗

Solution structure of the toluene 4-monooxygenase effector protein (T4moD).

Toluene 4-monooxygenase (T4MO) from Pseudomonas mendocina catalyzes the NADH- and O(2)-dependent hydroxylation of toluene to form p-cresol. The complex consists of an NADH oxidoreductase (T4moF), a Rieske ferredoxin (T4moC), a diiron hydroxylase [T4moH, with (alphabetagamma)(2) quaternary structure], and a catalytic effector protein (T4moD). The solution structure of the 102-amino acid T4moD effector protein has been determined from 2D and 3D (1)H, (13)C, and (15)N NMR spectroscopic data. The structural model was refined through simulated annealing by molecular dynamics in torsion angle space (DYANA software) with input from 1467 experimental constraints, comprising 1259 distance constraints obtained from NOEs, 128 dihedral angle constraints from J-couplings, and 80 hydrogen bond constraints. Of 60 conformers that met the acceptance criteria, the 20 that best satisfied the input constraints were selected to represent the solution structure. With exclusion of the ill-defined N- and C-terminal segments (Ser1-Asn11 and Asp99-Met102), the atomic root-mean-square deviation for the 20 conformers with respect to the mean coordinates was 0.71 A for the backbone and 1.24 A for all non-hydrogen atoms. The secondary structure of T4moD consists of three alpha-helices and seven beta-strands arranged in an N-terminal betaalphabetabeta and a C-terminal betaalphaalphabetabetabeta domain topology. Although the published NMR structures of the methane monooxygenase effector proteins from Methylosinus trichosporium OB3b and Methylococcus capsulatus (Bath) have a similar secondary structure topology, their three-dimensional structures differ from that of T4moD. The major differences in the structures of the three effector proteins are in the relative orientations of the two beta-sheets and the interactions between the alpha-helices in the two domains. The structure of T4moD is closer to that of the methane monooxygenase effector protein from M. capsulatus (Bath) than that from M. trichosporium OB3b. The specificity of T4moD as an effector protein was investigated by replacing it in reconstituted T4MO complexes with effector proteins from monooxygenases from other bacterial species: Pseudomonas pickettii PKO1 (TbuV, toluene 3-monooxygenase); Pseudomonas species JS150 (TbmC, toluene 2-monooxygenase); and Burkeholderia cepacia G4 (S1, toluene 2-monooxygenase). The results showed that the closely related TbuV effector protein (55% sequence identity) provided partial activation of the complex, whereas the more distantly related TbmC (34% sequence identity) and S1 (29% sequence identity) did not. The (1)H NMR chemical shifts of the side-chain amide protons of Asn34, a conserved, structurally relevant amino acid, were found to be similar in spectra of effector proteins T4moD and TbuV but not in the spectrum of TbmC. This suggests that the region around Asn34 may be involved in structural aspects contributing to functional specificity.

Amino Acid Sequence↗

Identification of potential regulatory sites of the Na+,K+-ATPase by kinetic analysis.

Kinetic models are presented that allow the Na(+),K(+)-ATPase steady-state turnover number to be estimated at given intra- and extracellular concentrations of Na(+), K(+), and ATP. Based on experimental transient kinetic data, the models utilize either three or four steps of the Albers-Post scheme, that is, E(2) --> E(1), E(1) --> E(2)P (or E(1) --> E(1)P and E(1)P --> E(2)P), and E(2)P --> E(2), which are the major rate-determining steps of the enzyme cycle. On the time scale of these reactions, the faster binding steps of Na(+), K(+), and ATP to the enzyme are considered to be in equilibrium. Each model was tested by comparing calculations of the steady-state turnover from rate constants and equilibrium constants for the individual partial reactions with published experimental data of the steady-state activity at varying Na(+) and K(+) concentrations. To provide reasonable agreement between the calculations and the experimental data, it was found that Na(+)/K(+) competition for cytoplasmic binding sites was an essential feature required in the model. The activity was also very dependent on the degree of K(+)-induced stimulation of the reverse reaction E(1) --> E(2). Taking into account the physiological substrate concentrations, the models allow the most likely potential sites of short-term Na(+),K(+)-ATPase regulation to be identified. These were found to be (a) the cytoplasmic Na(+) and K(+) binding sites, via changes in Na(+) or K(+) concentration or their dissociation constants, (b) ATP phosphorylation (as a substrate), via a change in its rate constant, and (c) the position of the E(2)<==>E(1) equilibrium.

Animals↗

Topology scanning and putative three-dimensional structure of the extracellular binding domains of the apical sodium-dependent bile acid transporter (SLC10A2).

The apical sodium-dependent bile acid transporter (ASBT, SLC10A2) facilitates the enterohepatic circulation of bile salts and plays a key role in cholesterol metabolism. The membrane topology of ASBT was initially scanned using a consensus topography analysis that predominantly predicts a seven transmembrane (TM) domain configuration adhering to the "positive inside" rule. Membrane topology was further evaluated and confirmed by N-glycosylation-scanning mutagenesis, as reporter sites inserted in the putative extracellular loops 1 and 3 were glycosylated. On the basis of a 7TM topology, we built a three-dimensional model of ASBT using an approach of homology-modeling and remote-threading techniques for the extramembranous domains using bacteriorhodopsin as a scaffold for membrane attachment points; the model was refined using energy minimizations and molecular dynamics simulations. Ramachandran scores and other geometric indicators show that the model is comparable in quality to the crystal structures of similar proteins. Simulated annealing and docking of cholic acid, a natural substrate, onto the protein surface revealed four distinct binding sites. Subsequent site-directed mutagenesis of the predicted binding domain further validated the model. This model agrees further with available data for a pathological mutation (P290S) because the mutant model after in silico mutagenesis loses the ability to bind bile acids.

Amino Acid Sequence↗

Real-time kinetic studies on the interaction of transforming growth factor alpha with the epidermal growth factor receptor extracellular domain reveal a conformational change model.

Transforming growth factor alpha (TGF-alpha), epidermal growth factor (EGF), and related factors mediate their biological effects by binding to the extracellular domain of the EGF receptor, which leads to activation of the receptor's cytoplasmic tyrosine kinase activity. Much remains to be determined, however, about the detailed molecular mechanism involved in this ligand-induced receptor activation. The determination of the binding mechanism and the related thermodynamic and kinetic parameters are of prime importance. To do so, we have used a surface plasmon resonance-based biosensor (the BIAcore) that allows the real-time recording of the interaction between TGF-alpha and the extracellular domain of the EGF receptor. By immobilizing different biotinylated derivatives of TGF-alpha on the sensor chip surface, we demonstrated that the N-terminus of TGF-alpha is not directly involved in receptor binding. By optimizing experimental conditions and interpreting the biosensor results by several data analysis methods, we were able to show that the data do not fit a simple binding model. Through global analysis of the data using a numerical integration method, we tested several binding mechanisms for the TGF-alpha/EGF receptor interaction and found that a conformational change model best fits the biosensor data. Our results, combined with other analyses, strongly support a receptor activation mechanism in which ligand binding results in a conformation-driven exposure of a dimerization site on the receptor.

Binding, Competitive↗

Models of (ZrO2)n complexes intercalated into montmorillonite.

The article discusses the properties of several model zirconium dioxide complexes (ZrO(2))(n)() intercalated into the interlayer space of montmorillonite clay. Grand canonical Monte Carlo simulation was used in a series of numerical experiments during analysis of the low-temperature nitrogen adsorption in the micropores thus generated. The goal of such experiments was to determine the geometrical parameters of introduced molecular complexes of different types inside micropores of various widths. The obtained information was used to characterize textural and structural properties of three pillared interlayer materials prepared by using pillaring species synthesized via aging of zirconyl chloride solutions containing as additives chlorides of Ca, Sr, or Ba. It was found that in the cases of Ba and Ca the interlayer micropores are filled with isolated tetramers (ZrO(2))(4). Meanwhile, the presence of Sr in the pillaring solution, most likely, favors the preservation of larger sheetlike complexes (ZrO(2))(8).

Journal Article↗

Competing through-space and through-bond, intramolecular triplet-energy transfer in a supposedly rigid ruthenium(II) tris(2,2'-bipyridine)--fullerene molecular dyad.

A ditopic ruthenium(II) tris(2,2'-bipyridyl)-based fullerene conjugate has been synthesized so as to separate the photoactive terminals by way of a short ethynylene spacer group that is expected to act as a rigid rod. Intramolecular triplet-energy transfer from the metal complex to the fullerene is quantitative at all temperatures and there is no indication for competing electron transfer. Temperature dependence studies indicate two pathways for triplet-energy transfer. An activationless route dominates at low temperature and is attributed to through-bond electron exchange that takes place via super-exchange interactions. The triplet energy of the bridging unit lies well above that of the metal complex. An activated process is switched-on at high temperatures and is believed to involve through-space electron exchange within closed conformations. Molecular dynamics simulations predict that, in addition to an extended conformation, the linker can distort in such a way that the terminals come into orbital contact. In fact, the resultant closed conformation possesses an idealised geometry for fast electron exchange.

Journal Article↗

Quinolinic acid is extruded from the brain by a probenecid-sensitive carrier system: a quantitative analysis.

Although the neurotoxic tryptophan-kynurenine pathway metabolite quinolinic acid originates in brain by both local de novo synthesis and entry from blood, its concentrations in brain parenchyma, extracellular fluid, and CSF are normally below blood values. In the present study, an intraperitoneal injection of probenecid (400 mg/kg), an established inhibitor of acid metabolite transport in brain, into gerbils, increased quinolinic acid concentrations in striatal homogenates, CSF, serum, and homogenates of kidney and liver. Direct administration of probenecid (10 mM) into the brain compartment via an in vivo microdialysis probe implanted into the striatum also caused a progressive elevation in both quinolinic acid and homovanillic acid concentrations in the extracellular fluid compartment but was without effect on serum quinolinic acid levels. A model of microdialysis transport showed that the elevations in extracellular fluid quinolinic acid and homovanillic acid levels following intrastriatal application are consistent with probenecid block of a microvascular acid transport mechanism. We conclude that quinolinic acid in brain is maintained at concentrations below blood levels largely by active extrusion via a probenecid-sensitive carrier system.

Animals↗

Theory relating in vitro and in vivo microdialysis with one or two probes.

In this paper, we further develop the general theory of microdialysis by extending the linear model of Bungay et al. to provide a theoretical basis for in vitro and in vivo microdialysis. Specifically, we considered the effect of active clearance processes on in vivo microdialysis, and thereby elaborated the theory of Benveniste et al. to endogenous compounds. We examined the use of steady state tissue diffusion resistance with negligible clearance processes to interpret microdialysis data. The influence of the tissue properties on the in vitro and in vivo recoveries in dual-probe microdialysis was analyzed and we simulated the effect of the operating parameters on dual probe microdialysis performance. We estimated that the minimum clearance rate constant detectable by microdialysis in a quasi-steady state is about 5.5 x 10(-5) s(-1). This minimum rate constant establishes a criterion, below which inhibition of the active clearance processes does not show detectable influences on the microdialysis extraction efficiency.

Agar↗

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