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Angiogenesis and osteogenesis in an orthopedically expanded suture.

The purpose of this study was to examine the angiogenic and the subsequent osteogenic responses during a 96-hour time-course after sutural expansion. Fifty rats were divided into: (1) a control group that received only angiogenic induction through injection of 5 ng/gm recombinant human endothelial cell growth factor (rhECGF); (2) an experimental group that received orthopedic expansion and rhECGF; (3) a sham group that received expansion and sodium chloride (NaCl) injection; and (4) a baseline group that received no expansion or injection. All rats were injected with 3H-thymidine (1.0 microCi/gm) 1 hour before death to label the DNA of S-phase cells. Demineralized sections (4 microm thick) were stained with hematoxylin and eosin. Angiogenesis and cell migration were analyzed with a previously established cell kinetics model. Analysis of variance was used to test the hypothesis that enhancement of angiogenesis stimulates reestablishment of osteogenic capability. Blood vessel number, area, and endothelial cell-labeled index significantly increased in experimental groups, but no difference was found between control and baseline groups. Labeled-pericyte index and activated pericyte numbers in the experimental group were also higher than in the sham groups. These results show that supplemental rhECGF enhances angiogenesis in expanded sutures but not in nonexpanded sutures. Data also suggest that pericytes are the source of osteoblasts in an orthopedically expanded suture.

Analysis of Variance↗

SO and SO2 in mass-loss envelopes of red giants: probes of nonequilibrium circumstellar chemistry and mass-loss rates.

We have made a search for SO in the mass-loss envelopes of 24 mostly oxygen-rich red giants, using the 8(8)-7(7) (344.3 GHz), 6(5)-5(4) (219.9 GHz), 4(3)-3(2) (138.2 GHz), and 3(2)-2(1) (99.3 GHz) transitions. One or more of these transitions were detected toward 14 envelopes, including those of the supergiants VY CMa and IRC +10420. The SO envelope of IK Tau was measurably extended, and that of IRC +10420 marginally so, allowing us to determine the envelope sizes directly. The SiO (v = 0) J = 8-7, J = 5-4, and J = 3-2 lines at 347.3, 217.1, and 130.3 GHz were detected simultaneously with the 344.3, 219.9, and 138.2 GHz SO lines, respectively. From our SO data, as well as from SO2 data published elsewhere (for a few sources), we derive the circumstellar SO and SO2 abundances and compare them with theoretical predictions. We find that the photospheric abundances of SO and SO2 are insignificant, and that SO and SO2 must be produced in the circumstellar envelopes. However, the (average) SO abundances are larger than even the peak values predicted by nonequilibrium circumstellar chemistry models. The large SO abundances imply that S is not heavily depleted into grains in these envelopes, and further suggest that (i) the stellar mass-loss rates in these objects are substantially larger than accepted values, (ii) the circumstellar H/H2 abundance ratio is quite small, and (iii) the reaction of O and SH dominates over the reaction of S and OH in the production of SO. In the peculiar bipolar nebula OH 231.8+4.2, the combined SO and SO2 abundance exceeds the cosmic S abundance, if the mass-loss rate in this object is smaller than or equal to l0(-4) M solar yr-1. In IK Tau the data can only be fitted with models in which the SO is distributed in a hollow shell, as predicted by the nonequilibrium chemistry models. In another three, which have no measurable OH, the discovery of SO is difficult to explain by such models.

Astronomical Phenomena↗

Skeletal muscle calcineurin: influence of phenotype adaptation and atrophy.

Calcineurin (CaN) has been implicated as a signaling molecule that can transduce physiological stimuli (e.g., contractile activity) into molecular signals that initiate slow-fiber phenotypic gene expression and muscle growth. To determine the influence of muscle phenotype and atrophy on CaN levels in muscle, the levels of soluble CaN in rat muscles of varying phenotype, as assessed by myosin heavy chain (MHC)-isoform proportions, were determined by Western blotting. CaN levels were significantly greater in the plantaris muscle containing predominantly fast (IIx and IIb) MHC isoforms, compared with the soleus (predominantly type I MHC) or vastus intermedius (VI, contains all 4 adult MHC isoforms). Three months after a complete spinal cord transection (ST), the CaN levels in the VI muscle were significantly reduced, despite a significant increase in fast MHC isoforms. Surprisingly, the levels of CaN in the VI were highly correlated with muscle mass but not MHC isoform proportions in ST and control rats. These data demonstrate that CaN levels in skeletal muscle are highly correlated to muscle mass and that the normal relationship with phenotype is lost after ST.

Adaptation, Physiological↗

Making it without losing it: Type A, achievement motivation, and scientific attainment revisited.

In a study by Matthews, Helmreich, Beane, and Lucker (1980), responses by academic psychologists to the Jenkins Activity Survey for Health Prediction (JAS), a measure of the Type A construct, were found to be significantly, positively correlated with two measures of attainment, citations by others to published work and number of publications. In the present study, JAS responses from the Matthews et al. sample were subjected to a factor analysis with oblique rotation and two new subscales were developed on the basis of this analysis. The first, Achievement Strivings (AS) was found to be significantly correlated with both the publication and citation measures. The second scale, Impatience and Irritability (I/I), was uncorrelated with the achievement criteria. Data from other samples indicate that I/I is related to a number of health symptoms. The results suggest that the current formulation of the Type A construct may contain two components, one associated with positive achievement and the other with poor health.

Achievement↗

Application of a data-mining method based on Bayesian networks to lesion-deficit analysis.

Although lesion-deficit analysis (LDA) has provided extensive information about structure-function associations in the human brain, LDA has suffered from the difficulties inherent to the analysis of spatial data, i.e., there are many more variables than subjects, and data may be difficult to model using standard distributions, such as the normal distribution. We herein describe a Bayesian method for LDA; this method is based on data-mining techniques that employ Bayesian networks to represent structure-function associations. These methods are computationally tractable, and can represent complex, nonlinear structure-function associations. When applied to the evaluation of data obtained from a study of the psychiatric sequelae of traumatic brain injury in children, this method generates a Bayesian network that demonstrates complex, nonlinear associations among lesions in the left caudate, right globus pallidus, right side of the corpus callosum, right caudate, and left thalamus, and subsequent development of attention-deficit hyperactivity disorder, confirming and extending our previous statistical analysis of these data. Furthermore, analysis of simulated data indicates that methods based on Bayesian networks may be more sensitive and specific for detecting associations among categorical variables than methods based on chi-square and Fisher exact statistics.

Algorithms↗

Blood vessel adaptation to gravity in a semi-arboreal snake.

The effects of vasoactive agonists on systemic blood vessels were examined with respect to anatomical location and gravity acclimation in the semi-arboreal snake, Elaphe Obsoleta. Major blood vessels were reactive to putative neurotransmitters, hormones or local factors in vessel specific patterns. Catecholamines, adenosine triphosphate, histamine and high potassium (80 mM) stimulated significantly greater tension per unit vessel mass in posterior than anterior arteries. Anterior vessels were significantly more sensitive to catecholamines than midbody and posterior vessels. Angiotensin II stimulated significantly greater tension in carotid artery than in midbody and posterior dorsal aorta. Arginine vasotocin strongly contracted the left and right aortic arches and anterior dorsal aorta. Veins were strongly contracted by catecholamines, high potassium and angiotensin II, but less so by adenosine triphosphate, arginine vasotocin and histamine. Precontracted vessel were relaxed by acetylcholine and sodium nitroprusside, but not by atrial natriuretic peptide or bradykinin. Chronic exposure of snakes to intermittent hypergravity stress ( + 1.5 Gz at tail) did not affect the majority of vessel responses. These data demonstrate that in vitro tension correlates with that catecholamines, as well as other agonists, are important in mediating vascular responses to gravitational stresses in snakes.

Adaptation, Physiological↗

Sensory characterization of a ready-to-eat sweetpotato breakfast cereal by descriptive analysis.

The sweetpotato [Ipomoea batatas (L.) Lam], an important industry in the United States, has been selected as a candidate crop to be grown on future long-duration space missions by NASA. Raw sweetpotato roots were processed into flour, which was used to formulate ready-to-eat breakfast cereal (RTEBC). Twelve trained panelists evaluated the sensory attributes of the extruded RTEBC using descriptive analysis. The samples were significantly different (P<0.05) for all attributes. Twelve perceived sensory attributes, which could be used to differentiate the appearance, texture, and flavor of sweetpotato RTEBC, were described. The data could be used to optimize the RTEBC and for designing studies to test its consumer acceptance.

Adolescent↗

Running performance and cardiovascular capacity are not impaired in creatine-depleted rats.

Several published reports have indicated that derangement of the phosphocreatine/creatine (Cr) energy-buffering system via Cr analogue feeding results in cardiomyopathy when cardiac performance is assessed in vitro. The present study was designed to examine indexes of cardiac performance in rats that have been chronically Cr depleted. Adult (180 +/- 4 g) rats were assigned to a normal diet (ND) (n = 8) or a Cr-depletion diet (CD) group (n = 10). After 61 +/- 1 days of ad libitum feeding, measurements of steady-state exercise O2 consumption were made. Hemodynamic indexes were then assessed during incremental running to peak sustained levels. Rats were then killed and the left ventricle was excised. In the CD group Cr was depleted 82% and V1 isomyosin decreased while V2 increased. O2 consumption during steady-state running was not different in CD rats. The respiratory exchange ratios of CD rats reflected a bias toward fat utilization during the latter stages of prolonged exercise. The exercise heart rates and peak systolic blood pressures of CD rats were slightly lower than those of ND rats. Both negative and positive rates of left ventricular pressure development were significantly reduced at all running speeds in the CD rats. CD rats were capable of exercise performance equal to that of ND animals. The hemodynamic and metabolic data suggest that the adaptations seen in the CD animals may be similar to those reported after endurance training. These results indicate that chronic Cr depletion does not impair either the circulatory or exercise capacity of rodents.

Animals↗

Shape-dependent control of cell growth, differentiation, and apoptosis: switching between attractors in cell regulatory networks.

Development of characteristic tissue patterns requires that individual cells be switched locally between different phenotypes or "fates;" while one cell may proliferate, its neighbors may differentiate or die. Recent studies have revealed that local switching between these different gene programs is controlled through interplay between soluble growth factors, insoluble extracellular matrix molecules, and mechanical forces which produce cell shape distortion. Although the precise molecular basis remains unknown, shape-dependent control of cell growth and function appears to be mediated by tension-dependent changes in the actin cytoskeleton. However, the question remains: how can a generalized physical stimulus, such as cell distortion, activate the same set of genes and signaling proteins that are triggered by molecules which bind to specific cell surface receptors. In this article, we use computer simulations based on dynamic Boolean networks to show that the different cell fates that a particular cell can exhibit may represent a preprogrammed set of common end programs or "attractors" which self-organize within the cell's regulatory networks. In this type of dynamic network model of information processing, generalized stimuli (e.g., mechanical forces) and specific molecular cues elicit signals which follow different trajectories, but eventually converge onto one of a small set of common end programs (growth, quiescence, differentiation, apoptosis, etc.). In other words, if cells use this type of information processing system, then control of cell function would involve selection of preexisting (latent) behavioral modes of the cell, rather than instruction by specific binding molecules. Importantly, the results of the computer simulation closely mimic experimental data obtained with living endothelial cells. The major implication of this finding is that current methods used for analysis of cell function that rely on characterization of linear signaling pathways or clusters of genes with common activity profiles may overlook the most critical features of cellular information processing which normally determine how signal specificity is established and maintained in living cells.

Animals↗

Chromosome aberrations induced by high-LET radiations.

Measurements of chromosome aberrations in peripheral blood lymphocytes are currently the most sensitive and reliable indicator of radiation exposure that can be used for biological dosimetry. This technique has been implemented recently to study radiation exposures incurred by astronauts during space flight, where a significant proportion of the dose is delivered by high-LET particle exposure. Traditional methods for the assessing of cytogenetic damage in mitotic cells collected at one time point after exposure may not be suitable for measuring high-LET radiation effects due to the drastic cell cycle perturbations and interphase cell death induced by this type of exposure. In this manuscript we review the recent advances in methodology used to study high-LET induced cytogenetic effects and evaluate the use of chemically-induced Premature Chromosome Condensation (PCC) as an alternative to metaphase analysis. Published data on the cytogenetic effects of in vitro exposures of high-LET radiation is reviewed, along with biodosimetry results from astronauts after short or long space missions.

Astronauts↗

Acid-growth response and alpha-expansins in suspension cultures of bright yellow 2 tobacco.

The possibility that Bright Yellow 2 (BY2) tobacco (Nicotiana tabacum L.) suspension-cultured cells possess an expansin-mediated acid-growth mechanism was examined by multiple approaches. BY2 cells grew three times faster upon treatment with fusicoccin, which induces an acidification of the cell wall. Exogenous expansins likewise stimulated BY2 cell growth 3-fold. Protein extracted from BY2 cell walls possessed the expansin-like ability to induce extension of isolated walls. In western-blot analysis of BY2 wall protein, one band of 29 kD was recognized by anti-expansin antibody. Six different classes of alpha-expansin mRNA were identified in a BY2 cDNA library. Northern-blot analysis indicated moderate to low abundance of multiple alpha-expansin mRNAs in BY2 cells. From these results we conclude that BY2 suspension-cultured cells have the necessary components for expansin-mediated cell wall enlargement.

Amino Acid Sequence↗

Mutagenic effects of a single and an exact number of alpha particles in mammalian cells.

One of the main uncertainties in risk estimation for environmental radon exposure using lung cancer data from underground miners is the extrapolation from high- to low-dose exposure where multiple traversal is extremely rare. The biological effects of a single alpha particle are currently unknown. Using the recently available microbeam source at the Radiological Research Accelerator Facility at Columbia University, we examined the frequencies and molecular spectrum of S1- mutants induced in human-hamster hybrid (A(L)) cells by either a single or an exact number of alpha particles. Exponentially growing cells were stained briefly with a nontoxic concentration of Hoechst dye for image analysis, and the location of individual cells was computer-monitored. The nucleus of each cell was irradiated with either 1,2,4, or 8 alpha particles at a linear energy transfer of 90 keV/microm consistent with the energy spectrum of domestic radon exposure. Although single-particle traversal was only slightly cytotoxic to A(L) cells (survival fraction approximately 0.82), it was highly mutagenic, and the induced mutant fraction averaged 110 mutants per 10(5) survivors. In addition, both toxicity and mutant induction were dose-dependent. Multiplex PCR analysis of mutant DNA showed that the proportion of mutants with multilocus deletions increased with the number of particle traversals. These data provide direct evidence that a single a particle traversing a nucleus will have a high probability of resulting in a mutation and highlight the need for radiation protection at low doses.

Alpha Particles↗

Neutron yields from 435 MeV/nucleon Nb stopping in Nb and 272 MeV/nucleon Nb stopping in Nb and Al.

Neutron fluences were measured from 435 MeV/nucleon Nb ions stopping in a Nb target and 272 MeV/nucleon Nb ions stopping in targets of Nb and Al for neutrons above 20 MeV and at laboratory angles between 3 degrees and 80 degrees. The resultant spectra were integrated over angles to produce neutron energy distributions and over energy to produce neutron angular distributions. The total neutron yields for each system were obtained by integrating over the angular distributions. The angular distributions from all three systems are peaked forward, and the energy distributions from all three systems show an appreciable yield of neutrons with velocities greater than the beam velocity. Comparison of the total neutron yields from the two Nb + Nb systems suggests that the average neutron multiplicity decreases with decreasing projectile energy. Comparison of the total yields from the two 272 MeV/nucleon systems suggests that the total yields show the same dependence on projectile and target mass number as do total inclusive neutron cross sections. The data are compared with Boltzmann-Uehling-Uhlenbeck model calculations.

Aluminum↗

Quantification of scaling exponents and crossover phenomena in nonstationary heartbeat time series.

The healthy heartbeat is traditionally thought to be regulated according to the classical principle of homeostasis whereby physiologic systems operate to reduce variability and achieve an equilibrium-like state [Physiol. Rev. 9, 399-431 (1929)]. However, recent studies [Phys. Rev. Lett. 70, 1343-1346 (1993); Fractals in Biology and Medicine (Birkhauser-Verlag, Basel, 1994), pp. 55-65] reveal that under normal conditions, beat-to-beat fluctuations in heart rate display the kind of long-range correlations typically exhibited by dynamical systems far from equilibrium [Phys. Rev. Lett. 59, 381-384 (1987)]. In contrast, heart rate time series from patients with severe congestive heart failure show a breakdown of this long-range correlation behavior. We describe a new method--detrended fluctuation analysis (DFA)--for quantifying this correlation property in non-stationary physiological time series. Application of this technique shows evidence for a crossover phenomenon associated with a change in short and long-range scaling exponents. This method may be of use in distinguishing healthy from pathologic data sets based on differences in these scaling properties.

Heart Diseases↗

Detecting transient cognitive impairment with EEG pattern recognition methods.

This paper describes an initial evaluation of a new method for assessing transient states of cognitive impairment associated with intoxication or fatigue: neural network pattern recognition applied to features of the electroencephalogram (EEG) recorded from subjects performing a standardized task. Nine subjects performed a working memory task during an extended testing session occurring over the course of one night, and encompassing an alert baseline period, a state of mild acute intoxication, and a state of fatigue compounded by "hangover" or intoxication after-effects. Relative to the alert baseline, task performance was less accurate in the other test conditions, providing evidence of transient cognitive impairment. These states of impairment were associated with changes in spectral characteristics of the EEG. Neural network-based EEG pattern recognition techniques were used to develop and test detectors of these changes. Brief testing data samples originating from the alert baseline condition could be discriminated from those recorded during the state of acute intoxication with 98% accuracy (p < 0.0001), and from those recorded during the state of fatigue/hangover with 92% accuracy (p < 0.001). Furthermore, networks trained on data from a group of subjects were found to accurately classify data from test subjects who were not part of the training group. These results demonstrate the feasibility of using neurophysiological monitoring methods for detecting transient cognitive impairment.

Acute Disease↗

Molecular basis of the increase in invertase activity elicited by gravistimulation of oat-shoot pulvini.

An asymmetric (top vs. bottom) increase in invertase activity is elicited by gravistimulation in oat-shoot pulvini starting within 3 h after treatment. In order to analyze the regulation of invertase gene expression in this system, we examined the effect of gravistimulation on invertase mRNA induction. Total RNA and poly (A)+RNA, isolated from oat pulvini, and two oligonucleotide primers, corresponding to two conserved amino-acid sequences (NDPNG and WECPD) found in invertase from other species, were used for the polymerase chain reaction (PCR). A partial-length cDNA (550 base pairs) was obtained and characterized. There was a 52% deduced amino-acid sequence homology to that of carrot beta-fructosidase and a 48% homology to that of tomato invertase. Northern blot analysis showed that there was an obvious transient accumulation of invertase mRNA elicited by gravistimulation of oat pulvini. The mRNA was rapidly induced to a maximum level at 1 h following gravistimulation treatment and gradually decreased afterwards. The mRNA level in the bottom half of the oat pulvinus was significantly higher (five-fold) than that in the top half of the pulvinus tissue. The induction of invertase mRNA was consistent with the transient enhancement of invertase activity during the graviresponse of the pulvinus. These data indicate that the expression of the invertase gene(s) could be regulated by gravistimulation at the transcriptional and/or translational levels. Southern blot analysis showed that there were four genomic DNA fragments hybridized to the invertase cDNA. This suggests that an invertase gene family may exist in oat plants.

Amino Acid Sequence↗

Phylogenetic analysis of the formin homology 2 domain.

Formin proteins are key regulators of eukaryotic actin filament assembly and elongation, and many species possess multiple formin isoforms. A nomenclature system based on fundamental features would be desirable, to aid the rapid identification and characterization of novel formins. In this article, we attempt to systematize the formin family by performing phylogenetic analyses of the formin homology 2 (FH2) domain, an independently folding region common to all formins, which alone can influence actin dynamics. Through database searches, we identify 101 FH2 domains from 26 eukaryotic species, including 15 in mice. Sequence alignments reveal a highly conserved yeast-specific insert in the "knob loop" region of the FH2 domain, with unknown functional consequences. Phylogenetic analysis using minimum evolution (ME), maximum parsimony (MP), and maximum likelihood (ML) algorithms strongly supports the existence of seven metazoan groups. Yeast FH2 domains segregate from all other eukaryotes, including metazoans, other fungi, plants, and protists. Sequence comparisons of non-FH2 regions support relationships between three metazoan groups (Dia, DAAM, and FRL) and examine previously identified coiled-coil and Diaphanous auto-regulatory domain sequences. This analysis allows for a formin nomenclature system based on sequence relationships, as well as suggesting strategies for the determination of biochemical and cellular activities of these proteins.

Adaptor Proteins, Signal Transducing↗

Vestibular stimulation leads to distinct hemodynamic patterning.

Previous studies demonstrated that responses of a particular sympathetic nerve to vestibular stimulation depend on the type of tissue the nerve innervates as well as its anatomic location. In the present study, we sought to determine whether such precise patterning of vestibulosympathetic reflexes could lead to specific hemodynamic alterations in response to vestibular afferent activation. We simultaneously measured changes in systemic blood pressure and blood flow (with the use of Doppler flowmetry) to the hindlimb (femoral artery), forelimb (brachial artery), and kidney (renal artery) in chloralose-urethane-anesthetized, baroreceptor-denervated cats. Electrical vestibular stimulation led to depressor responses, 8 +/- 2 mmHg (mean +/- SE) in magnitude, that were accompanied by decreases in femoral vasoconstriction (23 +/- 4% decrease in vascular resistance or 36 +/- 7% increase in vascular conductance) and increases in brachial vascular tone (resistance increase of 10 +/- 6% and conductance decrease of 11 +/- 4%). Relatively small changes (<5%) in renal vascular tone were observed. In contrast, electrical stimulation of muscle and cutaneous afferents produced pressor responses (20 +/- 6 mmHg) that were accompanied by vasoconstriction in all three beds. These data suggest that vestibular inputs lead to a complex pattern of cardiovascular changes that is distinct from that which occurs in response to activation of other types of somatic afferents.

Analysis of Variance↗