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The proportion and size of GABA-immunoreactive neurons in the magnocellular and parvocellular layers of the lateral geniculate nucleus of the rhesus monkey.

Neurons containing GABA-immunoreactivity in LGN of the macaque monkey were analyzed quantitatively in semithin (1 micron) sections. The percentage of GABA(+) cells per unit area of the sections was 26% in the magnocellular layers and 19% in the parvocellular layers. However, the percentage of GABA(+) cells in a unit volume of LGN, calculated by a stereological method that takes into account the observed difference in size of labeled and unlabeled somata, was 35% in the magnocellular layers and 25% in the parvocellular layers. GABA(+) somata in the magnocellular layers were significantly larger than those in the parvocellular layers. The possible role of GABAergic cells in inhibitory mechanisms of receptive fields of parvo- and magnocellular neurons are discussed in the light of current knowledge of the physiology and neural circuits of macaque LGN.

Animals↗

The marginal layer in the neocortex of a 7 week-old human embryo. A light and electron microscopic study.

Ultrastructural study of the molecular layer of the neocortex of a 7 week-old human embryo confirms recent observations on various laboratory animals that call for revision of some classical concepts of corticogenesis. 1. At 7 weeks, the subpial, marginal or molecular layer is the first layer to differentiate from the ventricular layer and represents almost half the thickness of the telencephalic vesicle. 2. The first cells that have already migrated from the ventricular zone, even before any cortical plate is visible, are to be found in this marginal layer. These large cells are well differentiated and most probably represent the so called Cajal Retzius cells. 3. The earliest synapses ever seen in human embryo are found in the marginal or plexiform layer; this indicates the presence of a precocious set-up for an elaborate neuronal circuitry at this level.

Cell Differentiation↗

Granular layer induction following the topical application of proliferating agents.

Evidence is provided for a possible dermal influence on the epidermis. Topical vitamin A stimulates a number of dermal cells with different enzyme reactions, and these invade the epidermis at about the time a granular layer is induced in mouse tail scale epidermis. N-hexadecane also induced a granular layer formation in the tail scale epidermis but the application of this agent only results in the invasion of the epidermis by non-specific esterase cells. These non-specific esterase cells are present in the follicular zone where a granular layer is usually present. It appears that dendritic cells may be responsible for the formation of a granular layer and that these cells in some way influence the keratinocytes to discharge their lyosomal enzymes and thus form a granular layer. It appears unlikely that the dendritic cells actually contribute their own acid hydrolases to the cell cytolysis necessary for the production of granular layer.

Acid Phosphatase↗

Surface properties from the S-layer of Clostridium thermosaccharolyticum D120-70 and Clostridium thermohydrosulfuricum L111-69.

Labelling experiments using a positively charged topographical marker for electron microscopy, polycationized ferritin, showed that the S-layers of two closely related clostridia Clostridium thermohydrosulfuricum L111-69 and C. thermosaccharolyticum D120-70 do not exhibit a net negative charge, as usually observed for bacterial cell surfaces. Chemical modification of reactive sites confirmed that amino and carboxyl groups are exposed on the S-layer surface of both strains. Amino-specific, bifunctional agents crosslinked both S-layer lattices. Studies with carbodiimides revealed that only the S-layer surface of C. thermohydrosulfuricum L111-69 had amino and carboxyl groups closely enough aligned to permit electrostatic interactions between the constituent protomers. The regular structure of this S-layer lattice was lost upon converting the carboxyl groups into neutral groups by amidation. Disintegration of both S-layer lattices occurred upon N-acetylation or N-succinylation of the free amino groups. Adhesion experiments showed that in neutral and weakly alkaline environment whole cells of C. thermosaccharolyticum D120-70 exhibited a stronger tendency to bind to charged surfaces than whole cells of C. thermohydrosulfuricum L111-69, but showed a lower tendency to bind to hydrophobic materials.

Cell Wall↗

Diffusion coefficients of oxygen and hemoglobin as obtained simultaneously from photometric determination of the oxygenation of layers of hemoglobin solutions.

The oxygenation of layers of deoxygenated hemoglobin solutions after a sudden exposure to a gas containing oxygen at a partial pressure P1 has been studied by a photometric method. Layer thicknesses varied between 50 and 250 micron, hemoglobin concentrations between 0.1 and 0.34kg/l, and oxygen partial pressures between 4.65 and 93.1 kPa (35 and 700 mmHg). The diffusion chamber containing the layer of hemoglobin solution permitted a step change in gas atmosphere without changing the optical apparatus constant. The following results were obtained: 1. The oxygen saturation increase is independent of the layer thickness when expressed as a function of time divided by layer thickness squared (normalized oxygenation time). This justifies the assumption of chemical equilibrium between oxygen and hemoglobin in the range considered. 2. The oxygen saturation increases proportionally to the square root of time over a wide range of oxygenation as expected. This range reaches to almost 100% oxygenation at P1=93.1kPa (700mmHg) but less far as P1 is lower. Thus at high P1 values there is a sharp boundary between the oxygenated and deoxygenated part of the layer allowing the application of the advancing front concept. 3. Fitting the theoretical equations derived in a preceding paper to the experimental results provides simultaneous values of the oxygen permeability (or, knowing oxygen solubility, of the oxygen diffusion coefficient) and of the hemoglobin diffusion coefficient. These values agree fairly well with values obtained by other authors from experiments yielding the diffusion coefficients of oxygen or hemoglobin separately.

Diffusion↗

Use of a wedge cuvette in thin layer photometry and its application to oximetry.

A wedge cuvette was constructed by fixing 2 glass plates at a known angle with a spacer at one end. This resulted in a thin layer with thickness varying from 0 to 250 micrometer. By measuring the intensity of a beam of light through the thin layer as a function of distance along the wedge (and thus layer thickness), the absorption coefficient at the light wavelength used could be obtained without a separate measurement of I0, the reference light intensity. In addition, the difficult problem of determining accurate layer thickness as encoutered in conventional thin layer photometry has been avoided. Tests of the wedge cuvette method with Evans Blue and Malachite Green serial dilutions as well as with haemoglobin solutions at several oxygen saturations demonstrate that accuracy of the order of 1% can be obtained. Application of the wedge cuvette in experiments on oxygen uptake by layers of haemoglobin solution are discussed.

Hemoglobins↗

[The tensile strength of bracket adhesives depending on the adhesive layer thickness--an in-vitro study].

The purpose of this study was to determine the influence of the thickness of the adhesive layer between bracket base and enamel surface on tensile bond strength. Stainless steel brackets with retentive net bases were bonded to bovine incisors with adhesive layers varying in thickness from a minimum of 0.0 mm to a maximum of 0.8 mm. Eight different orthodontic adhesives were investigated; in order to simulate the procedure of indirect bonding, combinations of two different adhesives were also tested. It was found that the highly filled composites provided greatest bond strength. Increasing the thickness of the adhesive layer had no significant influence on tensile bond strength. In the case of light-cured composites, maximum tensile bond strength was achieved at a thickness of the adhesive layer of 0.2 mm. This is probably due to better penetration of light at this thickness. In the case of chemically curing no-mix adhesives, it was impossible to produce effective adhesive layers thicker than 0.2 mm, presumably because curing at the primer paste interface becomes a problem at greater thickness. The influence of cement thickness on tensile bond strength was basically identical whether brackets were bonded with one or two adhesives. Thus, an additional adhesive-adhesive interface has no influence on bond strength. If adhesive layer thickness of more than 0.2 mm are required, a chemically cured, highly filled paste-paste system should be used.

Animals↗

Keratan sulfate content in the superficial and deep layers of osteophytic and nonfibrillated human articular cartilage in osteoarthritis.

Very thin slices of the superficial and deep layers of osteophytic and apparently normal articular cartilage from six human osteoarthritic femoral heads were digested with papain. The digests were analyzed for keratin sulfate content using an enzyme-linked immunosorbent assay (ELISA) with inhibition step, and for chondroitin sulfate and collagen contents using biochemical assays. Although there were marked differences in Safranin-O staining of the superficial and deep layers of osteophytic cartilage, these two layers had identical high ratios of chondroitin sulfate/collagen. Keratan sulfate was present only in small amounts in osteophytic cartilage. However, the deeper layer contained significantly more of this glycosaminoglycan. The deeper layer of articular cartilage contained approximately twice as much chondroitin sulfate and six times more keratan sulfate relative to collagen than the superficial layer. The results of this study showed that this new sensitive approach, which requires as little as 200 micrograms wet cartilage as starting material, provides important qualitative and quantitative information about the major constituents of the matrix.

Cartilage, Articular↗

Electrophoretic analysis of proteins and enantiomers using capillaries modified by a successive multiple ionic-polymer layer (SMIL) coating technique.

The applicability of three-layer coatings consisting of three different polymers (A(+)-B(-)-C(+) coating) prepared by a successive multiple ionic-polymer layer (SMIL) coating technique to the immobilization of polypeptides and/or proteins onto the inner surface of the capillaries was investigated to provide a high-performance separation medium for proteins and enantiomers in capillary electrophoresis (CE). To obtain a stable protein-coated capillary, high molecular mass poly(ethyleneimine) (PEI) was employed as the first layer in the A(+)-B(-)-C(+) coating, and then a cationic protein was immobilized as the third layer. Comparisons of analytical performances between the A(+)-B(-)-C(+) coating and the conventional SMIL-coated (A(+)-B(-)-A(+) coating) capillary were conducted. The CE separation of cationic proteins was successfully achieved with the prepared capillaries. In addition, the polypeptide- and protein-coated capillaries were applied to the chiral separation of a binaphthyl compound. It should be noted that the chiral separation efficiency was strongly dependent on the second anionic polymer layer of the coating. Effects of the interaction between oppositely charged ionic polymer layers on the separation efficiency are discussed.

Electrophoresis, Capillary↗

Structural and chemical characterisation of titanium deuteride films covered by nanoscale evaporated palladium layers.

Thin titanium deuteride (TiD(y)) films, covered by an ultra-thin palladium layer, have been compared with the corresponding titanium and palladium films using a combination of scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS). The TiD(y) layers were prepared under ultra-high vacuum (UHV) conditions by precisely controlled deuterium sorption at 298 K on a Ti film evaporated onto a Si(100) substrate. Both Ti and TiD(y) films were then covered in situ by a nanoscale Pd layer. It was found that a 10- to 12-nm-thick Pd layer protects the TiD(y) films efficiently against extensive air interaction. The morphology of both the surface and bulk Pd/TiD(y) (Ti) films have been observed using SEM and cross-sectional TEM analysis, respectively. A polycrystalline bulk morphology in both Ti and TiD(y) films accompanied by a fine-grained Pd surface was observed. High-magnification cross-sectional TEM images reveal the TiD(y) film to be plastically deformed leading to an increase in the roughness of the top Pd layer. Complex structures, including Moiré patterns, have been identified within the Pd/TiD(y) interface. The chemical nature of this interface has been analysed after partial sputtering of the Pd top layer using XPS. Besides TiD(y) and Pd, TiO and PdO were found to be the main chemical species in the interface region of the Pd/TiH(y) film. The XPS valence-band spectra of the Pd/TiD(y) interface reveal electronic features characteristic of a Pd-Ti bimetallic structure.

Journal Article↗

Activity in deep intermediate layer collicular neurons during interrupted saccades.

The activity of neurons located in the deep intermediate and adjacent deep layers (hereafter called just deep intermediate layer neurons) of the superior colliculus (SC) in monkeys was recorded during saccades interrupted by electrical stimulation of the brainstem omnipause neuron (OPN) region. The goal of the experiment was to determine if these neurons maintained their discharge during the saccadic interruption, and thus, could potentially provide a memory trace for the intended movement which ends accurately on target in spite of the perturbation. The collicular neurons recorded in the present study were located in the rostral three-fifths of the colliculus. Most of these cells tended to show considerable presaccadic activity during a delayed saccade paradigm, and, therefore, probably overlap with the population of SC cells called buildup neurons or prelude bursters in previous studies. The effect of electrical stimulation in the OPN region (which interrupted ongoing saccades) on the discharge of these neurons was measured by computing the percentage reduction in a cell's activity compared to that present during non-interrupted saccades. During saccade interruption about 70% of deep intermediate layer neurons experienced a major reduction (30% or greater) in their activity, but discharge recovered quickly after the termination of the stimulation as the eyes resumed their movement to finish the saccade on the target. Therefore, the pattern of activity recorded in most of the deep intermediate layer neurons during interrupted saccades qualitatively resembled that previously reported for the saccade-related burst neurons which tend to be located more dorsally in the intermediate layer. In contrast, some of our cells (30%) showed little or no perturbation in their activity caused by the saccade interrupting stimulation. Because all the more dorsally located burst neurons and the majority of our deep intermediate layer neurons show a total or major suppression in their discharge during interrupted saccades, it seems unlikely that the colliculus by itself could maintain an accurate memory of the desired saccadic goal or the remaining dynamic motor error required to account for the accuracy of the resumed movement which occurs following the interruption. However, it remains possible that the smaller proportion of our neurons whose activity was not perturbed during interrupted movements could play a role in the mechanisms underlying saccade accuracy in the interrupted saccade paradigm. Interrupted saccades have longer durations than normal saccades to the same target. Therefore, we investigated whether the discharge of our deeper collicular cells was also necessarily prolonged during interrupted saccades, and, if so, how the prolongation compared to the prolongation of the saccade. Sixty percent of our sample neurons showed a prolongation in discharge that was approximately the same as the prolongation in saccade duration (difference < 15 ms in magnitude). The, observation that temporal discharge in our neurons was perturbed to roughly match saccadic temporal perturbation suggests that dynamic feedback about ongoing saccadic motion is provided to the colliculus, but does not necessarily imply that this structure is the site responsible for the computation of dynamic motor error.

Animals↗

Probing the stability of S-layer-supported planar lipid membranes.

Isolated protein subunits of the crystalline bacterial cell surface layer (S-layer) of Bacillus coagulans E38-66 have been recrystallized on one side of planar black lipid membranes (BLMs) and their influence on the electrical properties, rupture kinetics and mechanical stability of the BLM was investigated. The effect on the boundary potential, the capacitance or the conductance of the membrane was negligible whereas the mechanical properties were considerably changed. The mechanical stability was characterized by applying voltage pulses or ramps to induce irreversible rupture. The amplitude of the voltage pulse leading to rupture allows conclusions on the ability of membranes to resist external forces. Surprisingly, these amplitudes were significantly lower for composite S-layer/lipid membranes compared to undecorated BLMs. In contrast, the delay time between the voltage pulse and the appearance of the initial defect was found to be drastically longer for the S-layer-supported lipid bilayer. Furthermore, the kinetics of the rupture process was recorded. Undecorated membranes show a fast linear increase of the pore conductance in time, indicating an inertia-limited defect growth. The attachment of an S-layer causes a slow exponential increase in the conductance during rupture, indicating a viscosity-determined widening of the pore. In addition, the mechanical properties on a longer time scale were investigated by applying a hydrostatic pressure across the BLMs. This causes the BLM to bulge, as monitored by an increase in capacitance. Compared to undecorated BLMs, a significantly higher pressure gradient has to be applied on the S-layer face of the composite BLMs to observe any change in capacitance.

Bacillus↗

Resuscitation of ischemically damaged pancreas by the two-layer (University of Wisconsin solution/perfluorochemical) mild hypothermic storage method.

We have shown that 24-hour preservation by a two-layer [University of Wisconsin solution (UW)/perfluorochemical (PFC)] cold storage method at 4 degrees C allowed tissue ATP synthesis and resuscitated canine pancreases subjected to 90 minutes of warm ischemia. The purpose of this study was to examine whether the two-layer (UW/PFC) mild hypothermic storage method at 20 degrees C could shorten a preservation period for recovery of ischemically damaged pancreas and clarify changes of tissue adenine nucleotide metabolism and tissue perfusions. After 90 minutes of warm ischemia, canine pancreas grafts were preserved by the two-layer method and then autotransplanted. Tissue adenine nucleotide levels at the end of preservation and tissue perfusions after reperfusion were measured. Pancreas grafts subjected to 90 minutes of warm ischemia did not survive (0 of 5), without preservation. During a 5-hour preservation by the two-layer cold storage method the grafts did not synthesize enough ATP to repair damaged cell, although tissue perfusions were maintained after reperfusion. Consequently, ischemically damaged pancreases were not resuscitated (0 of 3). However, during 5-hour preservation by the two-layer mild hypothermic storage method, the grafts supplied enough ATP for processes that repair damaged cells, and tissue perfusions were maintained after reperfusion. As a result, ischemically damaged grafts were resuscitated (5 of 5). We conclude that 5-hour preservation by the two-layer mild hypothermic storage method accelerates ATP synthesis, which is essential for repairing damaged cells and protects the vascular microcirculation. This method can resuscitate ischemically damaged pancreas faster and holds promise for pancreas-kidney transplantation from cardiac arrest donors.

Adenine Nucleotides↗

[Retinal nerve fiber layer thickness and peripapillary blood flow in glaucoma patients and healthy probands].

BACKGROUND: In progressive glaucoma there is increasing loss of retinal nerve fibers and therefore decreasing nerve fiber layer thickness. As measurements of capillary blood flow have been reported to depend on nerve fiber layer thickness, this could result in incorrectly high blood flow measurements in patients with advanced glaucoma. METHODS: In 33 healthy controls and 59 glaucoma patients we measured retinal nerve fiber layer thickness by laser polarimetry and relative capillary blood flow by scanning laser doppler flowmetry three times on the nasal and temporal peripapillary retina. For statistical analysis a regression analysis was used. RESULTS: The correlation coefficients for volume, velocity, and flow with nerve fiber layer thickness at the same location were 0.02/-0.03/-0.02 in the temporal retina and -0.22/-0.07/-0.19 in the nasal retina (all correlations nonsignificant). CONCLUSION: No correlation was found between nerve fiber layer thickness and capillary blood flow. Measurement of capillary blood flow in glaucoma patients thus does not appear to be affected by decreasing nerve fiber layer thickness.

Adolescent↗

Number of cell layers of the stratum corneum in normal skin - relationship to the anatomical location on the body, age, sex and physical parameters.

We counted the number of cell layers in the stratum corneum (SC) of normal skin taken from different anatomical locations of the body of 301 individuals of various ages. Frozen 6 microm thick sections were stained with a 1% aqueous solution of safranin and observed under a microscope after application of 2% KOH solution. There were great variations in the number of SC cell layers (mean +/- SD) according to location and among different individuals. The smallest number was found in genital skin (6 +/- 2), followed in order by skin of the face (9 +/- 2), neck (10 +/- 2), scalp (12 +/- 2), trunk (13 +/- 4), extremities (15 +/- 4) and the palms and soles (47 +/- 24). The heel showed the largest number (86 +/- 36). No definite correlation was found between the number of corneocyte layers and sex of the individual, whereas there was a slight increase in the number of SC layers with age in the skin of the cheek and back, particularly in male individuals. Comparison of these data with those from functional assessments of the SC of the skin from various locations of healthy adults showed that transepidermal water loss, an indicator of SC barrier function, reflected the number of corneocyte cell layers. In contrast, high-frequency conductance, an indicator of the hydration state of the outer SC, did not seem to be under the influence simply of the number of SC cell layers.

Abdomen↗

Basal layer reactivity in hyperplastic and metaplastic lesions of the bronchi as studied by lectin histochemistry.

The relationship between squamous metaplasia and squamous cell carcinoma of the bronchi has been the subject of controversy. We investigated basal cell hyperplasia, stratification, squamous metaplasia, and squamous cell carcinoma by means of lectin histochemistry with peanut agglutinin (PNA), Ulex europeus agglutinin-I, soybean agglutinin, and Dolichos biflorus agglutinin for all normal basal cells that are reactive. A basal layer stained with PNA was observed in basal cell hyperplasia, stratification, and squamous metaplasia, but this layer was not exhibited by squamous cell carcinoma. In hyperplasia and metaplasia, PNA staining was biased toward the lowest (basal) layer, whereas staining for the other lectins was more uniformly distributed across the layers. A PNA-positive basal layer may be important for the morphologic reversibility of the bronchial mucosa in hyperplastic and metaplastic lesions, and destruction of this layer may be associated with a progression from metaplasia to squamous cell carcinoma.

Adenocarcinoma↗

Detection in situ and characterization of lignin in the G-layer of tension wood fibres of Populus deltoides.

The occurrence of lignin in the additional gelatinous (G-) layer that differentiates in the secondary wall of hardwoods during tension wood formation has long been debated. In the present work, the ultrastructural distribution of lignin in the cell walls of normal and tension wood fibres from poplar (Populus deltoides Bartr. ex Marshall) was investigated by transmission electron microscopy using cryo-fixation-freeze-substitution in association with immunogold probes directed against typical structural motifs of lignin. The specificity of the immunological probes for condensed and non-condensed guaiacyl and syringyl interunit linkages of lignin, and their high sensitivity, allowed detection of lignin epitopes of definite chemical structures in the G-layer of tension wood fibres. Semi-quantitative distribution of the corresponding epitopes revealed the abundance of syringyl units in the G-layer. Predominating non-condensed lignin sub-structures appeared to be embedded in the crystalline cellulose matrix prevailing in the G-layer. The endwise mode of polymerization that is known to lead to these types of lignin structures appears consistent with such an organized cellulose environment. Immunochemical labelling provides the first visualization in planta of lignin structures within the G-layer of tension wood. The patterns of distribution of syringyl epitopes indicate that syringyl lignin is deposited more intensely in the later phase of fibre secondary wall assembly. The data also illustrate that syringyl lignin synthesis in tension wood fibres is under specific spatial and temporal regulation targeted differentially throughout cell wall layers.

Animals↗

Buckling of inner cell wall layers after manipulations to reduce tensile stress: observations and interpretations for stress transmission.

The inner layer of the cell wall in tissues that are under tensile stress in situ, e.g. epidermis and collenchyma of etiolated sunflower hypocotyls, shows a pattern of transverse folds when the tissues are detached and plasmolysed. This can be observed by Nomarski imaging of inner surfaces of the outer cell walls and electron microscopy of longitudinal sections after peeling the epidermis and bathing it in plasmolysing solutions. The folds are apparently caused by buckling of the inner layer due to the longitudinal compressive force exerted on this layer by the outer wall layer, when it shrinks after the removal of the longitudinal tensile stresses. In these stresses, two components can be distinguished: the tissue stress, disappearing on peeling, and that caused directly by turgor pressure, disappearing in hyperosmotic solution. Investigation of the buckling indicates that the outer layer of the cell wall transmits in situ most of the longitudinal tensile stress in the wall. The common concept that the inner layer of the wall is the region bearing most stress and therefore regulating growth can still be valid with respect to the transverse stress component.

Cell Wall↗