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Correlation of Pectin Methylesterase Activity in Root Caps of Pea with Root Border Cell Separation.

We tested predictions of the hypothesis that pectin methylesterase in the root cap plays a role in cell wall solubilization leading to separation of root border cells from the root tip. Root cap pectin methylesterase activity was detected only in species that release large numbers of border cells daily. In pea (Pisum sativum) root caps, enzyme activity is correlated with border cell separation during development: 6-fold more activity occurs during border cell separation than after cell separation is complete. Higher levels of enzyme activity are restored by experimental induction of border cell separation. A corresponding increase in transcription of a gene encoding root cap pectin methylesterase precedes the increase in enzyme activity. A dramatic increase in the level of soluble, de-esterified pectin in the root tip also is correlated with pectin methylesterase activity during border cell development. This increase in acidic, de-esterified pectin during development occurs in parallel with a decrease in cell wall/apoplastic pH of cells in the periphery of the root cap.

Journal Article↗

Phase Separation of Plant Cell Wall Polysaccharides and Its Implications for Cell Wall Assembly.

Concentrated binary mixtures of polymers in solution commonly exhibit immiscibility, resolving into two separate phases each of which is enriched in one polymer. The plant cell wall is a concentrated polymer assembly, and phase separation of the constituent polymers could make an important contribution to its structural organization and functional properties. However, to our knowledge, there have been no published reports of the phase behavior of cell wall polymers, and this phenomenon is not included in current cell wall models. We fractionated cell walls purified from the pericarp of unripe tomatoes (Lycopersicon esculentum) by extraction with cyclohexane diamine tetraacetic acid (CDTA), Na2CO3, and KOH and examined the behavior of concentrated mixtures. Several different combinations of fractions exhibited phase separation. Analysis of coexisting phases demonstrated the immiscibility of the esterified, relatively unbranched pectic polysaccharide extracted by CDTA and a highly branched, de-esterified pectic polysaccharide present in the 0.5 N KOH extract. Some evidence for phase separation of the CDTA extract and hemicellulosic polymers was also found. We believe that phase separation is likely to be a factor in the assembly of pectic polysaccharides in the cell wall and could, for example, provide the basis for explaining the formation of the middle lamella.

Journal Article↗

Dipole separability in a neuromagnetic source analysis.

By studying the ability of a one-dipole model to explain the magnetic field actually resulting from two dipoles, minimum requirements for a successful separation of two dipoles were explored. Two dipoles in different depths generally require a much higher signal-to-noise ratio (SNR) than two dipoles in the same depth. For the latter condition, the dipole distance as well as the angles between the moments and the line connecting the dipole locations (connecting line) were systematically varied. A perpendicular orientation of the two dipoles turned out to be the most favorable condition: the minimum distance required for a separation of two dipoles was more than four times smaller than for a configuration with both moments oriented parallel to the connecting line. Separability of parallel dipoles was moderately enhanced if both moments assumed an orientation perpendicular to the connecting line. The separability of two antiparallel dipoles is not limited by concurrence with a one-dipole model, but by the low signal amplitudes resulting from a mutual cancellation of the fields arising from the two dipoles, and by concurrence with a quadrupole model. The results are presented so that quantitative conclusions about dipole separability can be derived for arbitrary SNR's. The study does not generally disprove the common believe that magnetoencephalography has a relatively poor spatial resolution, but it qualifies this view by suggesting that under favorable conditions two sources with a distance of only 1 cm may be resolvable.

Electromagnetic Fields↗

Real-time separation of discontinuous adventitious sounds from vesicular sounds using a fuzzy rule-based filter.

The separation of pathological discontinuous adventitious sounds (DAS) from vesicular sounds (VS) is of great importance to the analysis of lung sounds since DAS are related to certain pulmonary pathologies. An automated way of revealing the diagnostic character of DAS, by isolating them from VS, based on their nonstationarity, is presented in this paper. The proposed algorithm uses two adaptive network-based fuzzy inference systems to compose a generalized fuzzy rule-based stationary-nonstationary filter (GFST-NST). The training procedure of the fuzzy inference systems involves the outputs of the wavelet transform-based stationary-nonstationary filter (WTST-NST), proposed by Hadjileontiadis and Panas [1]. The basic idea of the GFST-NST was initially proposed by the authors with the introduction of the fuzzy rule-based stationary-nonstationary filter (FST-NST) [2], tested with the separation of crackles from VS. The main contribution of this paper is the modification of the structure of the FST-NST filter to a serial-type fuzzy filter that, unlike the parallel operation of the FST-NST filter, sends a predicted stationary signal (VS) into the predictor of the nonstationary (DAS). Applying the GFST-NST filter to fine-coarse crackles and squawks, selected from three lung sound databases, the coherent structure of DAS is revealed and they are separated from VS. The separation performance of the GFST-NST filter was evaluated through quantitative and qualitative indexes that proved its efficiency and superiority against the FST-NST filter. When compared to the WTST-NST filter, the GFST-NST filter performed similarly in accuracy and objectiveness, but in a faster way. Thus, the GFST-NST filter combines the separation accuracy of the WTST-NST filter with the real-time implementation of the FST-NST filter, so it can easily be used in clinical medicine as a module of an integrated intelligent patient evaluation system.

Auscultation↗

Thermolysin treatment: a new method for dermo-epidermal separation.

The epidermis of superficial human skin samples could easily be separated from the dermis following incubation at +4 degrees C for 1 h in a solution containing 250-500 micrograms/ml thermolysin, a proteolytic enzyme hitherto mostly used for protein analysis. Light and electron microscopy revealed that the dermo-epidermal separation occurred at the basement membrane between the sites of bullous pemphigoid antigen and laminin and that the hemidesmosomes were selectively disrupted. The cohesion and morphology of the separated epidermis as well as the immunologic parameters investigated were not altered by this procedure. The clear cut dermo-epidermal separation produced by thermolysin treatment differed from the separation obtained with trypsin, which predominantly occurred between basal and suprabasal cells by disruption of desmosomes.

Adult↗

Pre-anesthetic maternal separation increases pups' locomotor behavior during emergence from anesthesia in rats.

BACKGROUND: Problematic behavior at emergence from anesthesia in children, partly linked with maternal separation, is a major problem in pediatric anesthesia. In humans, as well as in many other mammalian species, such separation causes psychological and behavioral changes. This study was to investigate whether or not pre-anesthetic maternal separation has a similar effect on rat pups. METHODS: This study was conducted on 66 3-week-old Sprague-Dawley male rats. The rats were divided into two groups; pups housed with a dam (n = 33) and those housed without (n = 33). Pups were anesthetized with 1.2% halothane for 30 min. Afterwards we recorded their locomotor behavior at emergence from general anesthesia using a video tracking system. RESULTS: Pre-anesthetic maternal separation significantly increased the maximum velocity and the distance traveled by pups at the emergence. CONCLUSION: Pre-anesthetic maternal separation influenced pups' locomotive behavior at emergence.

Anesthesia↗

Quantification of variability and uncertainty using mixture distributions: evaluation of sample size, mixing weights, and separation between components.

Variability is the heterogeneity of values within a population. Uncertainty refers to lack of knowledge regarding the true value of a quantity. Mixture distributions have the potential to improve the goodness of fit to data sets not adequately described by a single parametric distribution. Uncertainty due to random sampling error in statistics of interests can be estimated based upon bootstrap simulation. In order to evaluate the robustness of using mixture distribution as a basis for estimating both variability and uncertainty, 108 synthetic data sets generated from selected population mixture log-normal distributions were investigated, and properties of variability and uncertainty estimates were evaluated with respect to variation in sample size, mixing weight, and separation between components of mixtures. Furthermore, mixture distributions were compared with single-component distributions. Findings include: (1). mixing weight influences the stability of variability and uncertainty estimates; (2). bootstrap simulation results tend to be more stable for larger sample sizes; (3). when two components are well separated, the stability of bootstrap simulation is improved; however, a larger degree of uncertainty arises regarding the percentiles coinciding with the separated region; (4). when two components are not well separated, a single distribution may often be a better choice because it has fewer parameters and better numerical stability; and (5). dependencies exist in sampling distributions of parameters of mixtures and are influenced by the amount of separation between the components. An emission factor case study based upon NO(x) emissions from coal-fired tangential boilers is used to illustrate the application of the approach.

Journal Article↗

Anomalous binding characteristics of human thyroxine binding globulin due to a dilution-dependent separation artefact.

Contrary to the accepted view, a recent study using Sephadex column separation suggested that thyroxine binding globulin (TBG) binds T4 and T3 with similar affinity, but with a much larger capacity for T4 than T3. We have evaluated this finding by comparing this separation method with equilibrium dialysis, taking account of the effect of serum dilution with each method. Estimates of free T4 fraction by equilibrium dialysis (with magnesium chloride precipitation) were valid over a wide range of serum dilutions. In contrast, Sephadex column separation gave a major overestimate of free hormone (underestimate of binding) in less diluted serum, indicating that this method cannot be used to establish a value for T4 affinity independent of serum dilution. Such a systematic error will result in a greater underestimate of affinity for the ligand with higher affinity when two ligands are compared at a single serum dilution. By equilibrium dialysis at 37 degrees C, the affinity of T4 for TBG was approximately 13-fold higher than that of T3, while the capacity of TBG for both T4 and T3 was close to the concentration of immunoreactive TBG. The previous report of similar T4 and T3 affinities appears to be due to a dilution-dependent underestimate of T4 affinity inherent in Sephadex column separation. Direct comparison of binding kinetics of various ligands requires a separation method that is valid over a wide range of binding protein concentrations.

Alpha-Globulins↗

The chromosome partitioning proteins Soj (ParA) and Spo0J (ParB) contribute to accurate chromosome partitioning, separation of replicated sister origins, and regulation of replication initiation in Bacillus subtilis.

Soj (ParA) and Spo0J (ParB) of Bacillus subtilis belong to a conserved family of proteins required for efficient plasmid and chromosome partitioning in many bacterial species. Unlike most Par systems, for which intact copies of both parA and parB are required for the Par system to function, inactivating soj does not cause a detectable chromosome partitioning phenotype whereas inactivating spo0J leads to a 100-fold increase in the production of anucleate cells. This suggested either that Soj does not function like other ParA homologues, or that a cellular factor might compensate for the absence of soj. We found that inactivating smc, the gene encoding the structural maintenance of chromosomes (SMC) protein, unmasked a role for Soj in chromosome partitioning. A soj null mutation dramatically enhanced production of anucleate cells in an smc null mutant. To look for effects of a soj null on other phenotypes perturbed in a spo0J null mutant, we analysed replication initiation and origin positioning in (soj-spo0J)+, Deltasoj, Deltaspo0J and Delta(soj-spo0J) cells. All of the mutations caused increased initiation of replication and, to varying extents, affected origin positioning. Using a new assay to measure separation of the chromosomal origins, we found that inactivating soj, spo0J or both led to a significant defect in separating replicated sister origins, such that the origins remain too close to be spatially resolved. Separation of a region outside the origin was not affected. These results indicate that there are probably factors helping to pair sister origin regions for part of the replication cycle, and that Soj and Spo0J may antagonize this pairing to contribute to timely separation of replicated origins. The effects of Deltasoj, Deltaspo0J and Delta(soj-spo0J) mutations on origin positioning, chromosome partitioning and replication initiation may be a secondary consequence of a defect in separating replicated origins.

Bacillus subtilis↗

Rapid separation of enkephalins and endorphins on Sep-Pak reverse phase cartridges.

A simple, rapid method is described for separating neuropeptides. Samples were injected on Sep-Pak disposable reverse phase cartridges and eluted with a step gradient of n-propanol in formate-pyridine buffer. Enkephalins are separated from metabolic products. Separation alpha, beta and gamma-endorphin demonstrated, and Met- and Leu-enkephalin are also separated. This inexpensive, reproducible method a useful in studies of neuropeptide metabolism, purification of synthesized peptides, separation prior to RIA, or concentration of neuropeptides.

Chromatography↗

Long-term follow-up of previously separated pigtail macaques: group and individual differences in response to novel situations.

A follow-up study of monkeys who experienced maternal separations as infants 2.5-4.9 years earlier was performed. Matched pairs of previously separated and nonseparated control monkeys were observed individually in four unfamiliar situations, and their behavioral responses and plasma cortisol levels were measured. Results indicated that previously separated animals displayed more disturbance than did their matched controls, and levels of disturbance were significantly correlated with the degree of behavioral and physiological disturbance displayed during the original maternal separations. The results support the idea that the perception of the maternal separation may be a significant factor in whether long-term behavioral deficits exist in social and emotional functioning.

Affective Symptoms↗

Computational simulation of the blood separation process.

The aim of this work is to construct a computational fluid dynamics model capable of simulating the quasitransient process of apheresis. To this end a Lagrangian-Eulerian model has been developed which tracks the blood particles within a delineated two-dimensional flow domain. Within the Eulerian method, the fluid flow conservation equations within the separator are solved. Taking the calculated values of the flow field and using a Lagrangian method, the displacement of the blood particles is calculated. Thus, the local blood density within the separator at a given time step is known. Subsequently, the flow field in the separator is recalculated. This process continues until a quasisteady behavior is reached. The simulations show good agreement with experimental results. They shows a complete separation of plasma and red blood cells, as well as nearly complete separation of red blood cells and platelets. The white blood cells build clusters in the low concentrate cell bed.

Animals↗

Effects of electrode-myocardial separation on cardiac stimulation in conductive solution.

INTRODUCTION: Effects of a conductive bath and electrode-myocardial separation on cardiac stimulation have not been elucidated. These factors may play a role in endocardial catheter stimulation or defibrillation. METHODS AND RESULTS: We studied effects of a bath and separation on transmembrane voltage changes during stimulation (deltaVm) and excitation thresholds in rabbit hearts, cultured rat cardiac cell monolayers, and cardiac bidomain computer models. Similar to previous epicardial measurements with no bath, a dogbone pattern of deltaVm during stimulation was found in bathed epicardium and right ventricular septal endocardium and in models of bathed anisotropic myocardium. Electrode-myocardial separation altered spatial distributions of deltaVm, moved reversals of the sign of deltaVm farther from the stimulation epicenter, and decreased aspect ratio of deltaVm (i.e., length/width of dogbone contours of deltaVm). The separation increased thresholds and reduced maximal deltaVm, while deltaVm at sites away from maxima increased or decreased. Anodal thresholds in models initially were larger than those in experiments and decreased when models were altered to include nonuniform cellular coupling. Existence of nonuniformity in monolayers was indicated by irregular excitation patterns. CONCLUSION: Electrode-myocardial separation alters spatial distributions of deltaVm, which may impact on arrhythmia induction by altering distributions of states of deltaVm-sensitive ion channels. The results also indicate that excitation thresholds may depend on tissue nonuniformities.

Animals↗

Macronuclear development in conjugants of Tetrahymena thermophila, which were artificially separated at meiotic prophase.

Conjugant pairs of Tetrahymena thermophila were mechanically separated by vigorous pipetting at the early stages of meiotic prophase. The complete sequence of conjugational nuclear events including the appearance of pronuclei, development of the new macronuclei (postzygotic development), and resorption of the old macronuclei was observed in the separated cells, without pronuclear exchange. The pronuclei in the separated cells were recognised by the presence of components of the extranuclear cytoskeleton, which were labelled with anti-tubulin and anti-fenestrin antibodies in the same way as in undisturbed conjugants. The apical region of the separated conjugants (the post-junction area), corresponding to the junction area of conjugants was labelled with anti-fenestrin antibody and maintained the properties required for the nuclear development. The results of the genetic study were consistent with a hypothesis that cytogamy (pronuclear fusion) was induced in the separated conjugants. Therefore, the lasting cell contact is not necessary for the successful completion of conjugational nuclear events.

Animals↗

Short mother-infant separation during first week of life influences the duration of breastfeeding.

Thirty term infants, who were separated 1-6 days (mean 3.3 days) from their mothers during the first week of life because of minor illnesses, were compared with 116 term non-separated infants with regard to breastfeeding. At 3 months of age the frequency of entirely breastfed infants was 37% in the separated group versus 72% in the non-separated group, p less than 0.001. Thus a few days of separation between mothers and infants during the first week of life seems to decrease the duration of breastfeeding.

Breast Feeding↗

Influence of optical properties and fiber separation on laser doppler flowmetry.

Microcirculatory blood flow can be measured using a laser Doppler flowmetry (LDF) probe. However, the readings are affected by the tissue's optical properties (absorption and scattering coefficients, mu(a) and mu(s)) and probe geometry. In this study the influence of optical properties [mu(a)in(0.053,0.23) mm-1,mu(s)in(14.7,45.7) mm-1] on LDF perfusion and LDF sampling depth was evaluated for different fiber separations. In vitro measurements were made on a sophisticated tissue phantom with known optical properties that mimicked blood flow at different depths. Monte Carlo simulations were carried out to extend the geometry of the tissue phantom. A good correlation between measured and simulated data was found. The simulations showed that, for fixed flow at a discrete depth, the influence of mu(s) or mu(a) on LDF perfusion increased with an increase in flow depth and decreased with an increase in fiber separation. For a homogeneous flow distribution, however, the perfusion varied 40% due to variations in the optical properties, almost independent of the fiber separation (0.23-1.61 mm). Therefore, the effect in real tissue is likely to vary due to the unknown heterogeneous blood flow distribution. Further, the LDF sampling depth increased with a decrease in mu(s) or mu(a) and an increase in fiber separation. For fiber separation of 0.46 mm, the e-1 sampling depth ranged from 0.21 to 0.39 mm.

Blood Flow Velocity↗

The effects of spatial separation in distance on the informational and energetic masking of a nearby speech signal.

Although many studies have shown that intelligibility improves when a speech signal and an interfering sound source are spatially separated in azimuth, little is known about the effect that spatial separation in distance has on the perception of competing sound sources near the head. In this experiment, head-related transfer functions (HRTFs) were used to process stimuli in order to simulate a target talker and a masking sound located at different distances along the listener's interaural axis. One of the signals was always presented at a distance of 1 m, and the other signal was presented 1 m, 25 cm, or 12 cm from the center of the listener's head. The results show that distance separation has very different effects on speech segregation for different types of maskers. When speech-shaped noise was used as the masker, most of the intelligibility advantages of spatial separation could be accounted for by spectral differences in the target and masking signals at the ear with the higher signal-to-noise ratio (SNR). When a same-sex talker was used as the masker, the intelligibility advantages of spatial separation in distance were dominated by binaural effects that produced the same performance improvements as a 4-5-dB increase in the SNR of a diotic stimulus. These results suggest that distance-dependent changes in the interaural difference cues of nearby sources play a much larger role in the reduction of the informational masking produced by an interfering speech signal than in the reduction of the energetic masking produced by an interfering noise source.

Adult↗

Evaluation of two voice-separation algorithms using normal-hearing and hearing-impaired listeners.

Two signal-processing algorithms, designed to separate the voiced speech of two talkers speaking simultaneously at similar intensities in a single channel, were compared and evaluated. Both algorithms exploit the harmonic structure of voiced speech and require a difference in fundamental frequency (F0) between the voices to operate successfully. One attenuates the interfering voice by filtering the cepstrum of the combined signal. The other uses the method of harmonic selection [T. W. Parsons, J. Acoust. Soc. Am. 60, 911-918 (1976)] to resynthesize the target voice from fragmentary spectral information. Two perceptual evaluations were carried out. One involved the separation of pairs of vowels synthesized on static F0's; the other involved the recovery of consonant-vowel (CV) words masked by a synthesized vowel. Normal-hearing listeners and four listeners with moderate-to-severe, bilateral, symmetrical, sensorineural hearing impairments were tested. All listeners showed increased accuracy of identification when the target voice was enhanced by processing. The vowel-identification data show that intelligibility enhancement is possible over a range of F0 separations between the target and interfering voice. The recovery of CV words demonstrates that the processing is valid not only for spectrally static vowels but also for less intense time-varying voiced consonants. The results for the impaired listeners suggest that the algorithms may be applicable as components of a noise-reduction system in future digital signal-processing hearing aids. The vowel-separation test, and subjective listening, suggest that harmonic selection, which is the more computationally expensive method, produces the more effective voice separation.

Algorithms↗