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Membrane phospholipid phase separations in plants adapted to or acclimated to different thermal regimes.

The phase separation temperatures of total leaf phospholipids from warm and cool climate plants were determined in order to explore the relationship of lipid physical properties to a species' thermal habitat. The separation temperatures were determined by measuring the fluorescence intensity and fluorescence polarization of liposomes labeled with the polyene fatty acid probe trans-parinaric acid. To focus on a single climatic region, Mojave Desert dicots (chiefly ephemeral annuals) were examined, with plants grown under identical conditions whenever possible. Winter active species showed lower phase separation temperatures than the summer active species. A group of warm climate annual grasses showed separation temperatures distinctly higher than those of a group of cool climate grasses, all grown from seed under the same conditions. Growth at low temperature seems correlated with (and may require) a low phase separation temperature. Winter active ephemerals appear genetically programmed to synthesize a mixture of phospholipids which will not phase separate in the usual growth conditions. When the lipids of desert perennials were examined in cool and warm seasons, there was a pronounced seasonal shift in the phase separation temperature, implying environmental influences on lipid physical properties. The relationship of these results to high and low temperature tolerance is discussed.

Journal Article↗

An in vitro model of immune complex-mediated basement membrane zone separation caused by pemphigoid antibodies, leukocytes, and complement.

In this study, an in vitro model of immune complex-mediated basement membrane zone separation caused by periphigoid antibodies, serum complement, and peripheral blood leukocytes is described. When cryostat sections of fresh-frozen normal human skin were treated with either of 4 bullous pemiphigoid sera containing complement-activating anti-basement membrane zone antibodies and subsequently incubated at 37 degrees C with normal human peripheral blood leukocytes and fresh human serum, leukocytes attached to 96% of the basement membrane zone in 100% of sections. Sixty-seven percent of the sections developed focal areas of basement membrane zone separation resembling dermal-epidermal separation described in early pemphigoid lesions. In control sections in which either leukocytes, pemphigoid antibody or fresh human serum were omitted, significantly less leukocyte attachment and basement membrane zone separation occurred. Evidence that leukocytes caused separation was supported by an absolute requirement for viable leukocytes during incubation, a high correlation between leukocyte attachment and separation and experiments showing that leukocytes attached to the basement membrane zone were activated. This study provides the first in vitro evidence directly supporting a functional role for immune-complex mediated inflammation in the pathogenesis of basement membrane zone separation and blisters in bullous pemphigoid.

Antibodies↗

Involvement of FtsZ in coupling of nucleoid separation with septation.

The cell-cycle parameters of an Escherichia coli strain expressing essential division gene ftsZ at one-fifth of its normal level, because of antisense regulation by DicF RNA, have been analysed. Inhibition of FtsZ expression affects neither the generation time nor the replication initiation mass, the C period, or the constriction period, but it does dramatically retard the initiation of constriction relative to replication termination. Separation of the nucleoids is equally postponed, indicating that division is not coupled to termination of replication, but to partitioning. The severe inhibition of nucleoid separation by DicF RNA, and its suppression by overproduction of FtsZ, suggest a role for FtsZ in the control of separation, and consequently in the coupling of separation and division. We suggest that the normal pattern of nucleoid separation previously found in cells deficient in ftsZ function was a consequence of the loss of a negative effect exerted by FtsZ on separation. In agreement with this view, we find that nucleoid separation is temporarily inhibited after arrest of FtsZ synthesis, but is later resumed as FtsZ is further diluted into the elongating filaments.

Bacterial Proteins↗

Severing all ties between mother and daughter: cell separation in budding yeast.

At the end of nuclear division in the budding yeast, acto-myosin ring contraction and cytokinesis occur between mother and daughter cells. This is followed by cell separation, after which mother and daughter cells go their separate ways. While cell separation may be the last event that takes place between the two cells, it is nonetheless under tight regulation which ensures that both cells are viable upon separation. It is becoming increasingly clear that the components of the cell separation machinery are controlled at various levels, including the temporal and spatial regulation of the genes encoding for the components and the specific localization of the components to the neck. In addition, these regulatory controls are co-ordinated with exit from mitosis, thereby placing a mechanistic link between the end of mitosis and cell separation. More importantly, the success of the cell separation event is contingent upon the presence of a trilaminar septum, whose assembly is dependent on a host of proteins which localize to the neck over the span of one cell division cycle.

Cell Compartmentation↗

Dissociation in the effects of neonatal maternal separations on maternal care and the offspring's HPA and fear responses in rats.

The development of the hypothalamic-pituitary-adrenal (HPA) response to stress is influenced by the early mother-infant relationship. In rats, early handling (brief daily mother-offspring separations) attenuates the adult offspring's HPA and fear responses compared to both nonhandling (no separations) and maternal separation (prolonged daily separations). It has been proposed that variation in the amount of maternal care mediates these effects of neonatal manipulations on the adult offspring's stress and fear responses. Here we tested this hypothesis by assessing maternal care and the adult offspring's HPA and fear responses in Lister hooded rats which were subjected to either early handling (EH) or maternal separation (MS) from postnatal day 1-13, or were left completely undisturbed (nonhandled, NH) throughout this period. Both EH and MS induced a more active nursing style and elevated levels of maternal care compared to NH. Total levels of maternal care were indistinguishable between EH and MS, but diurnal distribution differed. MS dams showed elevated levels of maternal care following the 4-h separation period, thereby fully compensating for the amount of maternal care provided by EH dams during the time MS dams were separated from their pups. However, while EH resulted in reduced HPA and fear responses in the adult offspring compared to NH, MS and NH offspring did not differ. Our findings therefore demonstrate dissociation in the effects of EH and MS on maternal care and on the stress and fear responses in the offspring. This indicates that maternal care cannot be the sole mediator of these effects.

Animals↗

An ultrastructural comparison of dermo-epidermal separation techniques.

Dermo-epidermal separation through the lamina lucida is an essential technique for immunoblotting studies and for the diagnostic immunofluorescence of autoimmune bullous diseases. The most widely used methods of producing skin separation in the laboratory are suction blister induction and incubation in 1 molar sodium chloride. More recently the use of a proteolytic enzyme, thermolysin, has been described for this purpose. We examined the electron microscopic appearance of five suction blisters, five skin specimens separated by 1 M NaCl, and five treated with thermolysin. Both suction blister formation and treatment with 1 M NaCl resulted in a clean separation through the lamina lucida in all specimens examined. However specimens treated with thermolysin developed intra-epidermal separation in four cases without any lamina lucida separation in three. Suction blister formation was associated with hemidesmosome disruption. Incubation in 1 M NaCl remains the most reproducible, convenient, and reliable method of producing dermo-epidermal separation in the laboratory.

Basement Membrane↗

Separation surfaces for laminar flow in branching tubes--effect of Reynolds number and geometry.

Flow visualization experiments of Newtonian laminar flow through branching tubes have been performed to identify the shape of the separation surface or flow divider. The influences of Reynolds number, flow fraction into the side branch, and branch geometry on the separation surface shape have been considered. The shapes presented in this paper are formed by the intersection of the separation surface with the cross section of the parent tube. At low Re the separation surfaces are curved in a convex manner, bulging away from the opening of the side branch. Increasing Re causes the surface to become concave. At Re > 194 the surfaces can become closed for Q* > 0.3. The branch angle has no noticeable effect on the separation surface shape. The side to parent branch diameter ratio, Db/Dp, has a strong influence at low Re. As Re increases the diameter effect diminishes. Previous studies have shown no difference between the separation surfaces of T and Y type junctions at low Re. At Re = 194 there is a marked difference between the separation surfaces of T and Y bifurcations.

Biomechanical Phenomena↗

A one-dimensional unsteady separable and reattachable flow model for collapsible tube-flow analysis.

Taking into account both flow separation and reattachment observed in available experimental results on flows in a quasi-two-dimensional channel, we present a one-dimensional unsteady flow model, which is applicable to a flow in a collapsible tube. The flow model has been derived from the two-dimensional Navier-Stokes equations by introducing the concept of a dividing streamline, which divides a separated flow into a jet and a dead-water zone. We also present a criterion for the determination of a separation point. Numerical results show that the locations of the predicted separation points agree well with the experimental data. The predicted static pressure of the separated flow is almost constant downstream of the separation point and increases quickly just before the reattachment point as observed in the experiment. Finally, using the present flow model and the separation criterion, we examine the oscillatory behavior of an unsteady flow in a symmetric channel whose walls move sinusoidally.

Compliance↗

Separation of concurrent broadband sound sources by human listeners.

The effect of spatial separation on the ability of human listeners to resolve a pair of concurrent broadband sounds was examined. Stimuli were presented in a virtual auditory environment using individualized outer ear filter functions. Subjects were presented with two simultaneous noise bursts that were either spatially coincident or separated (horizontally or vertically), and responded as to whether they perceived one or two source locations. Testing was carried out at five reference locations on the audiovisual horizon (0 degrees, 22.5 degrees, 45 degrees, 67.5 degrees, and 90 degrees azimuth). Results from experiment 1 showed that at more lateral locations, a larger horizontal separation was required for the perception of two sounds. The reverse was true for vertical separation. Furthermore, it was observed that subjects were unable to separate stimulus pairs if they delivered the same interaural differences in time (ITD) and level (ILD). These findings suggested that the auditory system exploited differences in one or both of the binaural cues to resolve the sources, and could not use monaural spectral cues effectively for the task. In experiments 2 and 3, separation of concurrent noise sources was examined upon removal of low-frequency content (and ITDs), onset/offset ITDs, both of these in conjunction, and all ITD information. While onset and offset ITDs did not appear to play a major role, differences in ongoing ITDs were robust cues for separation under these conditions, including those in the envelopes of high-frequency channels.

Acoustic Stimulation↗

Distortion product otoacoustic emission (2f1-f2) amplitude as a function of f2/f1 frequency ratio and primary tone level separation in human adults and neonates.

Distortion product otoacoustic emission (DPOAE) (2f1-f2) amplitude is dependent upon both the frequency ratio and level separation of the eliciting primary tones. In adults it has been established that, on average, a f2/f1 ratio of 1.22 is optimal for evoking the most robust DPOAE; DPOAE amplitude is systematically reduced when f2/f1 ratio is either increased or decreased, thus forming a bandpass function (Harris et al., 1989). The frequency ratio function (DPOAE amplitude x f2/f1 ratio) is thought to reflect the filtering properties of the cochlea (Allen and Fahey, 1993; Brown et al., 1993). Primary tone level separation also influences DPOAE amplitude, with a 10- to 15-dB level difference between the primary tones (L1 > L2) typically generating largest amplitude in adults. Equivalent studies have not been conducted in neonates. The present study evoked the 2f1-f2 DPOAE in adults, term and premature neonates to define the optimal f2/f1 ratio and L1-L2 level separation and to investigate the filtering properties of the developing cochlea. Two f2 frequencies were investigated: 1500 and 6000 Hz. F2 was held constant while f1 was varied to produce 13 frequency ratios. Primary tone level separation varied from 15 to 0- in 5-dB intervals. ANOVA were conducted on the resulting f2/f1 frequency ratio and level separation data. Results showed that the mean optimal frequency ratio for DPOAE generation is comparable in adults and neonates. Also, either a 15- or 10-dB level separation (L1 > L2) produced the largest DPOAE amplitude for adults and term neonates whereas DPOAEs from premature neonates appeared to be relatively insensitive to primary tone level separation. The f2/f1 frequency ratio functions were similar in shape, slope, and bandwidth for adults and neonates, suggesting adult-like cochlear filtering prior to term birth. This finding is in agreement with previous work from our laboratory reporting adult-like DPOAE suppression tuning curves in term-born neonates [Abdala et al., Hear. Res. (1996)].

Acoustic Stimulation↗

Diagnostic significance of flow separation within the carotid bifurcation demonstrated by digital subtraction angiography.

The presence of an area of reversed blood flow due to flow separation in the interval carotid artery is a normal finding in Doppler ultrasound studies in vivo as well as in model carotid bifurcations. This flow separation phenomenon is caused by the geometry of the carotid bifurcation and the fluid dynamics of blood. We demonstrated the flow separation phenomenon on lateral-projection intra-arterial digital subtraction angiograms in 99 of 100 carotid bifurcations. The mean duration of flow separation was 5.8 seconds, with values up to 14 seconds in normal carotid bifurcations. The presence of this flow separation phenomenon is almost independent of atherosclerotic lesions and is not correlated with cerebral ischemia. Therefore, the angiographic finding of a flow separation has no diagnostic value. However, our results refer to a factor that may be important in the genesis of atherosclerosis at the carotid bifurcation, namely the duration of the stay of blood and its components within the area of flow separation.

Adult↗

Study on composition and particle characteristics of shredded municipal waste for the improvement of separation efficiency in a municipal bulky waste processing facility.

In order to propose a new and more efficient method of separation for shredded municipal bulky waste (SBW), particle characteristics (namely shape, density and terminal settling velocity) were investigated. It was found that particles of the combustibles such as wood, paper and plastics were included in the fraction smaller than 50 mm. Consequently, separation of SBW into combustibles and non-combustibles by particle size is difficult. Since the particles of the various materials are of all shapes, it is difficult to sort them by shape discrimination. The overall separation efficiencies using air flow and density for SBW separation were estimated and the maximum values are as high as approximately 90% and 99% respectively. However, because of the overlapping ranges of the terminal settling velocities in air for glass, metals, rubber, and plastics, the overall efficiency will become smaller if those materials are major components of the feed. It is concluded that density separation is more suitable for the separation of combustibles and non-combustibles in SBW than air separation.

Air Movements↗

Fat reduction in pericardial suction blood by spontaneous density separation: an experimental model on human liquid fat versus soya oil.

Pericardial suction blood (PSB) contains mediastinal liquid wound fat with an embolic potential to cause brain damage after cardiopulmonary bypass (CPB). The aims were to measure how fat separates spontaneously from blood by density and how temperature and fat surface adhesion affect the results under experimental conditions. Human liquid fat was heat-extracted from retrieved pericardial fat tissue of coronary artery bypass graft (CABG) patients (n = 10). Human fat or soya oil, 5% and 10%, respectively, were mixed with postoperatively shed mediastinal blood (n = 20). The mixture was loaded into a temperature-controlled (37 degrees C, 20 degrees C, 10 degrees C) vertical separation column. At 1, 2.5, 5 and 10 minutes, the blood was collected in five fractions, representing layers of density separation, followed by centrifugation. Human fat solidified at 8 degrees C. Soya oil remained liquid below 0 degrees C. Soya oil separated fast in water, but was slower in blood. At 10 minutes and 37 degrees C 73 +/- 6% of added soya oil was found in the top 20% fraction. Human fat at 37 degrees C behaved similarly to soya oil, with 58 +/- 2% separation at 10 minutes. However, at lower temperatures the density separation became less efficient (p < 0.001), whereas human fat more effectively adhered to the walls of the column, which added to the removal. In total, 66%-78% of the human fat was removed, depending on temperature. In conclusion, fat in PSB can be reduced by simple density separation and surface adhesion while it is temporarily retained from the CPB circuit.

Adipose Tissue↗

Separation of noncutaneous epithelia in a fetus diagnosed in utero with junctional epidermolysis bullosa.

This study was undertaken to identify the extent of involvement of cutaneous and noncutaneous epithelia during expression in utero of junctional epidermolysis bullosa. Skin and other organs from a 19-wk estimated gestational age fetus affected with junctional epidermolysis bullosa and from age-matched controls were examined by light and transmission electron microscopy. In the affected fetus, skin samples from different body regions including trunk, leg, arm, and finger all showed some separation at the dermal-epidermal junction in the plane of the lamina lucida. Hemidesmosomes were absent or hypoplastic, whereas anchoring fibrils appeared normal in structure and number. Interfollicular epidermis appeared to have separated easily, whereas some follicles remained anchored in the dermis. Areas of epithelium in the trachea and bronchi had separated, but within the lung parenchyma the epithelium of smaller bronchioles and alveoli remained attached to supporting connective tissue. The transitional epithelium of the urinary bladder showed small areas of separation compared with the gall bladder epithelium, which showed extensive separation. Gall bladder epithelium in several control fetuses also was consistently separated. In the affected fetus as well as in controls, gall bladder had multiple layers of basal lamina, a previously unrecognized structural feature of human fetal gall bladder. Epithelia that remained intact included the linings of the stomach and small and large intestines and endothelium of large and small vessels in all organs. Kidney, spleen, liver, and lymph nodes appeared normal in all aspects. In control samples, neither skin nor most of the noncutaneous epithelia had separated from the underlying connective tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Advancement and separation of apex hole eliminators with cementless Duraloc 100 cups. A report of nineteen cases.

BACKGROUND: From 1992 to 1996, we performed 541 cementless total hip arthroplasties using a Duraloc 100 acetabular component with an apex hole eliminator that did not have a positive stop design. During routine follow-up radiographic evaluation, we noted several hips with advanced or separated hole eliminators. This study presents the clinical consequences of this occurrence and its relationship to polyethylene wear and osteolysis. METHODS: A total of 541 hips (358 with a minimum duration of follow-up of five years) were evaluated clinically with use of the method of Merle D'Aubigné and Postel. Anteroposterior pelvic and iliac oblique radiographs were used to evaluate polyethylene wear, osteolysis, hole eliminator position, and component stability. RESULTS: Nineteen hips that had been followed for an average of eighty-nine months showed advancement or complete separation of the hole eliminator on radiographic evaluation. In most hips, advancement of the hole eliminator was visible on only the iliac oblique radiograph. In all but one hip with separation of the hole eliminator, complete plug separation was detectable on both the iliac oblique and anteroposterior pelvic radiographs. No association was found between polyethylene wear and either advancement or separation of the hole eliminator. No osteolysis was visible radiographically in the hips with hole eliminator advancement, but osteolysis was always visible in the hips that had complete separation of the hole eliminator. CONCLUSIONS: The hole eliminator has been redesigned since November 1995 to prevent possible advancement. However, patients with earlier designs should be followed carefully because advancement or separation can be expected in about 5% of the patients who have been followed for a minimum of five years. Despite this complication, which is seen radiographically, all patients were satisfied with the function of the hip. We do not recommend surgery solely to remove or replace the hole eliminator.

Adult↗

Freezability of equine semen after glass beads column separation.

REASONS FOR PERFORMING STUDY: The success rate of artificial insemination following the freezing of stallion semen is limited; therefore, improving the stallion semen quality after the freezing and thawing process is a necessary objective. OBJECTIVES: To investigate the influence of glass bead column separation on the freezability of stallion semen. HYPOTHESIS: Glass beads in a column separator remove damaged and dead spermatozoa in the ejaculate during centrifugation. METHODS: In total, 50 ejaculates from 6 Lipizzaner stallions were studied. Each ejaculate was divided into 2 parts, one half processed following standard procedure and the second half used for the column separation procedure. After freezing, semen quality was evaluated using standard tests for motility, morphology and viability of semen. RESULTS: Motility and progressive motility of the column-separated (CS) semen were significantly higher (P < 0.001) before freezing and immediately, 24 and 48 h after thawing. A significant increase (P < 0.001) in the percentage of hypoosmotic positive spermatozoa was observed in CS samples. The percentage of total morphological changes in the separated samples before and after freezing was significantly lower (P < 0.001) compared with samples prepared using the standard procedure. A substantial decrease (P < 0.001) was found in the percentage of spermatozoa with damaged acrosomes. However, the percentage of spermatozoa with coiled tails was increased in the separated samples (P < 0.001). CONCLUSIONS: Column separation before freezing has a positive effect on the quality of thawed equine semen. POTENTIAL RELEVANCE: The quality of CS frozen/thawed samples indicates their potential use for increasing insemination success in mares.

Animals↗

Separation of sutured tendon ends when different suture techniques and different suture materials are used. An experimental study in rabbits.

Calcaneal tendons in rabbits were divided and then sutured with varying suture techniques and suture materials. The separation of the tendon ends after suturing was registered with an X-ray technique--the tendon ends being identified by thin steel wires. Independently of the suture techniques and the suture materials a biphasic separation of the tendon ends was registered. The first separation took place during the first 4-5 days while the second separation began after 20-24 days. A small initial separation seems to indicate that the tendon suture used is adequate. The initial separation can therefore be used to evaluate a suturing technique and how the result is influenced by the suture material used. In the present study we could show that steel wire was ideal for a Bunnell suture. Prolene and Ethiflex cannot be recommended. A suture technique with a single short loop gave good results with all suture materials. Dexon ligatures placed around the tendon ends according to Schink in order to prevent the strands of the suture from cutting the tissue, did not reduce the initial separation. When a complex suture technique, like a Bunnell suture, is used the physical properties of the suture material influence the result. With a simple suture technique, a single short loop, good results can be achieved with every suture material.

Animals↗

Expanding the search for a linear separability constraint on category learning.

Formal models of categorization make different predictions about the theoretical importance of linear separability. Prior research, most of which has failed to find support for a linear separability constraint on category learning, has been conducted using tasks that involve learning two categories with a small number of members. The present experiment used four categories with three or nine patterns per category that were either linearly separable or not linearly separable. With overall category structure equivalent across category types, the linearly separable categories were found to be easier to learn than the not linearly separable categories. An analysis of individual participants' data showed that there were more participants operating under a linear separability constraint when learning large categories than when learning small ones. Formal modeling showed that an exemplar model could not account for many of these data. These results are taken to support the existence of multiple processes in categorization.

Exploratory Behavior↗