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PubMed1992

The superficial plexiform layer: a third retinal association area.

Electron microscopy enables the identification of a new association area in the mammalian retina. It is a region whose processes and synaptic junctions form a diffuse and intermittent layer bridging the boundary between the optic nerve fibre layer and ganglion cell layer. It involves displaced amacrine cells, retinal ganglion cells and probably interplexiform cell processes. Because it is close to the vitreal surface of the retina and has several properties of a plexiform layer, it has been named the superficial plexiform layer. It is much more sparse than the outer or inner plexiform layers in the rabbit, but contains a significant and substantial density of 6,100 synapses/mm2 near the visual streak. Morphological criteria distinguish two classes of synapses in the new association area. One has features of Colonnier's symmetric type and is formed by amacrine-like processes or their terminals, onto other amacrine-like processes or the cell bodies, dendrites and axon hillocks of some large ganglion cells. It makes up 79% of the population at a density of 4,800 synapses/mm2 in the examined sample. The second class arises from large processes of the fibre layer, which resemble axons rather than amacrine cell branches, has Colonnier's asymmetric form, and synapses onto a variety of other neuronal processes and cells. At a local density of 1,300/mm2 this type forms 21% of the population. A series of experiments including Wallerian degeneration, retrograde degeneration, electron microscopy and horseradish peroxidase (HRP) transport was employed to determine the nature of the large synapse-forming processes of the fibre layer that were the most amenable to investigation. It is concluded that the processes project into the optic nerve because they can be filled with HRP by retrograde transport from the end of the transected optic nerve. Although this result is based on limited evidence, it is complemented by the observation that the majority of the fibres undergo retrograde degeneration within 25 days of optic nerve section. It is concluded that the processes are either centrifugal fibres or the axons of retinal ganglion cells. The persistence of a relatively normal density of synapse-forming large fibres for seven days after optic nerve section, without Wallerian degeneration, argues that they are not the terminals of centrifugal axons. This conclusion is complemented by a separate class of profiles that did degenerate in this short period and are concluded to represent the centrifugal fibres. The large synapsing processes of the fibre layer are concluded to be the axons of some large ganglion cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

Pyramidal neurons in layer 5 of the rat visual cortex. I. Correlation among cell morphology, intrinsic electrophysiological properties, and axon targets.

Previous work has established two structure/function correlations for pyramidal neurons of layer 5 of the primary visual cortex of the rat. First, cells projecting to the superior colliculus have thick apical dendrites with a florid terminal arborization in layer 1, whereas those projecting to the visual cortex of the opposite hemisphere have thinner apical dendrites that terminate below layer 1, without a terminal tuft (e.g., Hallman et al.: J Comp Neurol 272:149, '90). Second, intracellular recording combined with dye injection has revealed two classes of cells: the first has a thick, tufted apical dendrite and fires a distinctive initial burst of two or more impulses, of virtually fixed, short interspike interval, in response to current injection; and the other, with a slender apical dendrite lacking a terminal tuft, tends to have a longer membrane time constant and higher input resistance, and does not fire characteristic bursts (e.g., Larkman and Mason: J Neurosci 10:1407, '90). The present study combined intracellular recording in isolated slices of rat visual cortex and injection of carboxyfluorescein, to reveal soma-dendritic morphology, with prior injection of rhodamine-conjugated microspheres into the superior colliculus or contralateral visual cortex to label neurons according to the target of their axons. This permitted a complete correlation of morphology, intrinsic electrophysiological properties, and identity of the projection target for individual pyramidal cells. Neurons retrogradely labeled from the opposite visual cortex were found in all layers except layer 1 while those labeled from the superior colliculus lay exclusively in layer 5. Within layer 5 interhemispheric cells were more concentrated in the lower half of the layer but extensively overlapped the distribution of corticotectal cells. Every cell studied that projected to the superior colliculus was of the bursting type and had a thick apical dendrite with a terminal tuft. Every cell in this study projecting to the opposite visual cortex was a "nonburster" and had a slender apical dendrite with fewer oblique branches that ended without a terminal tuft, usually in the upper part of layer 2/3. Interhemispheric cells also had rounder, less conical somata and generally had fewer basal dendrites than corticotectal neurons. Many cells with the physiological and morphological characteristics of interhemispheric cells were not back-labeled from the opposite visual cortex, implying that pyramidal cells of this type can have other projection targets (e.g., other cortical sites in the ipsilateral hemisphere).(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1995

Prenatal development of fibrous (white matter), protoplasmic (gray matter), and layer I astrocytes in the human cerebral cortex: a Golgi study.

The prenatal developmental histories of layer I, fibrous (white matter), and protoplasmic (gray matter) astrocytes have been studied in the human neocortex by the rapid Golgi method. The developmental route followed by each of these astrocytes is a distinct process which evolves from a specific precursor, occurs at a different time, and is linked to a specific event. The differentiation of layer I astrocytes is linked to the neocortex external glial limiting membrane (EGLM), that of fibrous astrocytes to the early white matter vascularization and maturation, and that of protoplasmic astrocytes to the late gray matter ascending vascularization and maturation. At the start of development, three glial precursors are established in the neocortex: 1) original radial neuroectodermal cells with nuclei above the primordial plexiform layer (PPL) by losing their ependymal and retaining their pial attachments become early astrocytes of layer I and EGLM components; 2) neuroectodermal cells with nuclei below the PPL that retain their pial and ependymal attachments become type I radial glial cells which are committed to the guidance of neurons and the early EGLM maintenance; and, 3) neuroectodermal cells that lose their pial but retain their ependymal attachment are transformed into type II radial glial precursors. By progressively losing their ependymal attachment, type II radial glia precursors become freely migrating cells, establish vascular contacts, and differentiate into fibrous astrocytes (and into oligodendrocytes?) throughout the subplate, developing white matter, and paraventricular regions. After the formation of the gray matter, additional layer I astrocytes are needed for the EGLM late prenatal and postnatal maintenance because type I radial glia cells start to regress and to reabsorb their EGLM endfeet. A late ependyma-to-pia migration of glial precursors progressively repopulates layer I with additional astrocytes and establishes the ephemeral subpial granular layer (SGL) of Ranke. From the 15th week of gestation to the time of birth, late astrocytes of layer I lose their EGLM attachments, migrate freely into the maturing gray matter, establish vascular contacts, and differentiate into protoplasmic astrocytes. The protoplasmic astrocytes of the gray matter evolve from transformation of layer I astrocytes rather than from radial glia cells as is generally believed.

Astrocytes↗
PubMed1987

The cell envelope of Thermoproteus tenax: three-dimensional structure of the surface layer and its role in shape maintenance.

The sulphur-dependent archaebacterium Thermoproteus tenax has a cylindrical cell shape variable in length, but constant in diameter. Its whole surface is covered by a regular protein layer (S-layer). The lattice has p6 symmetry and a lattice constant of 32.8 nm. The three-dimensional reconstruction from a tilt series of isolated and negatively stained S-layer shows a complex mass distribution of the protein: a prominent, pillar-shaped protrusion is located at the 6-fold crystallographic axis with radiating arms connecting neighbouring hexamers in the vicinity of the 3-fold axis. The base vectors of the S-layer lattice have a preferred orientation with respect to the longitudinal axis of the cell. The layer can be seen as a helical structure consisting of a right-handed, two-stranded helix, with the individual chains running parallel. Supposing that new S-layer protein is inserted at lattice faults (wedge disclinations) near the poles, growing of the layer would then proceed by moving a disclination at the end of the helix. The constant shape of the cell, as well as the particular structure of the layer, strongly suggest that this S-layer has a shape-maintaining function.

Journal Article↗
PubMed1992

Compositional dependence of calcium phosphate layer formation in fluoride Bioglasses.

Bioglasses form a double layer composed of apatite and a silica-rich layer when placed in a simulated physiological solution as well as in living tissue [A.E. Clark, C.G. Pantano, and L. L. Hench, "Auger spectroscopic analysis of bioglass corrosion films," J. Am. Ceram. Soc., 59(1-2), 37-39 (1976).]. In the present work, the mechanisms of the calcium phosphate layer and the silica-rich layer formation of fluoride Bioglasses in Tris-buffer solution are studied as a function of the SiO2 content. Fourier Transform Infrared Reflection Spectroscopy (FTIRS) is used to investigate the mechanism of formation of calcium phosphate and silica-rich layers on the glass surface. Ion concentration in reacted solution and elemental depth profiles are obtained by Induced Coupled Plasma Atomic Emission Spectrometry (ICP) and Auger Electron Spectroscopy (AES), respectively. Si--O bonds with one nonbridging oxygen and Si--O--Si bonds form at the early stage of reaction. Strong phosphorus ion uptake occurs when an amorphous calcium phosphate layer crystallizes. Glasses with high silica content (conventional glass) form the silica-rich layer first followed by a calcium phosphate layer on top. However, glasses with low silica content (invert glass) form both layers simultaneously. The rate of apatite formation decreases with increasing SiO2 content, especially in the region of conventional glass compositions. Ion release rates decreases as SiO2 content increases, with a significant change occurring at the compositional boundary between invert and conventional glasses.

Biocompatible Materials↗
PubMed2006

The outermost layer of egg-jelly is crucial to successful fertilization in the newt, Cynops pyrrhogaster.

The significance of egg-jelly layers in internal fertilization was evaluated in the newt, Cynops pyrrhogaster. In this species, six egg-jelly layers, J1, J2, J3, J4, J5 and the outermost J6 layers, are accumulated on the surface of the fertilizable eggs in pars convoluta of the oviduct. When a large number of sperm (about 6 x 10(5)) were placed on eggs having different numbers of jelly layers, all the eggs were fully fertilized, although many of the eggs developed abnormally. Upon insemination using about 600 sperm, only eggs with the full set of jelly layers were fertilized at a high rate with normal development. Since around 300 (the range of 48-1,192) sperm were observed on and in the egg-jelly in naturally spawned eggs, we conclude that the J6 layer must be present on the outermost surface of the egg-jelly for successful internal fertilization of the newt. Previous studies have suggested that the J6 layer is a prerequisite for the initiation of sperm motility and the acrosome reaction. In the present study, the fertilization rate decreased in eggs with a full set of jelly layers when inseminated using acrosome-reacted and motile sperm. However, the fertilization rate was high when motile sperm with intact acrosome was used. These results suggest that induction of the sperm acrosome reaction in the J6 layer is an important step in the internal fertilization of the newt.

Acrosome Reaction↗
PubMed2006

Magnetic resonance imaging of tissue and vascular layers in the cat retina.

PURPOSE: To report the visual resolution of multiple cell and vascular "layers" in the cat retina using MRI. MATERIALS AND METHODS: T2- and diffusion-weighted MRI at 4.7 Tesla was performed. Layer-specific thickness, T2, spin density, apparent diffusion coefficient perpendicular (ADC(perpendicular)) and parallel (ADC(parallel)) to the retinal surface were tabulated. T1-weighted MRI was acquired before and after intravenous administration of Gd-DTPA and subtraction images were obtained. Histology was performed for validation. RESULTS: Three distinct "layers" were observed. The inner strip nearest to the vitreous (exhibiting large T2, ADC, spin density with Gd-DTPA enhancement) overlapped the ganglion cell layer, bipolar cell layer, and the embedded retinal vascular layer. The middle strip (exhibiting small T2, ADC, spin density without Gd-DTPA enhancement) overlapped the photoreceptor cell layer and the inner and outer segments. The outer strip (exhibiting large T2, ADC, spin density with Gd-DTPA enhancement) overlapped the tapetum and choroidal vascular layer. T2, spin density, ADC(perpendicular) and ADC(parallel) of different "layers" were tabulated. The inner strip was slightly thicker than the other two strips. The total thickness, including neural and nonneural retina, was 358 +/- 13 microm (N = 6) by MRI and 319 +/- 77 microm (N = 5) by histology. CONCLUSION: MRI provides a noninvasive tool to study the retina with laminar specificity without depth limitation.

Animals↗
PubMed1994

Significance of viable skin layers in percutaneous permeation and its implication in mathematical models: theoretical consideration based on parameters for betamethasone 17-valerate.

The role of viable skin layers (viable epidermis and dermis) is examined by the three-layer model using parameter values for betamethasone 17-valerate. The mathematical three-layer diffusion model indicates that the lag time and half-life after vehicle removal in epidermis and split-thickness skin are longer than those in stratum corneum without viable layers, even if drug flux at steady-state is minimally altered. The theoretical values of the lag time and half-life predicted for epidermis and split-thickness-skin samples are compatible with the values observed in another in vitro study. The results predicted by the three-layer model indicate that the simplified model (single-layer or compartment model), which regards the whole skin as one diffusion layer or one compartment, may be warranted because all three skin layers have the same half-life after vehicle removal. The parameters used in the simplified model are estimated from some of the following values directly obtainable in the experiment: flux from skin and amount in the whole skin at steady-state, lag time, and drug concentration or amount maintained to be unchanged in the donor site. However, the simplified model often cannot resolve some discrepancy between the data and model (e.g., the ratio of the half-life to lag time) even if the data may be explained by the three-layer model.

Betamethasone Valerate↗
PubMed1998

The Denaturation of Lysozyme Layers Adsorbed at the Hydrophobic Solid/Liquid Surface Studied by Neutron Reflection.

We have studied the adsorption of lysozyme layers at a hydrophobic silicon water interface using specular neutron reflection. The hydrophobic surface was obtained by self-assembly of a densely packed monolayer of octadecyltrichlorosilane (OTS) onto the natural silica layer on the smooth surface of a (111) silicon block. The effect of pH on the adsorbed lysozyme layer was examined at a constant lysozyme concentration of 0.03 g dm-3 and at a constant ionic strength of 0.02 M. Reflectivity profiles at different pH show that adsorption is irreversible with respect to pH, the composition and structure of the final layer being dependent on the route by which the pH was achieved. The adsorbed protein layer was found to divide into approximately two regions, a densely packed thin layer next to the OTS surface and a diffuse thicker layer extending into the bulk solution. None of the dimensions of this structure corresponds to those of the globular protein in solution, suggesting that, unlike its adsorption at the hydrophilic silica/water interface, lysozyme is denatured at the OTS/water surface. The irreversible adsorption is explained by the combined interaction of the hydrophobic attraction of the hydrophobic fragments in lysozyme to the OTS surface and electrostatic repulsion within the adsorbed layer. The hydrophobic surface induces the exposure of hydrophobic fragments from the lysozyme assembly. The thickness of the dense layer suggests that the denatured protein adsorbs in the form of peptide chains with the hydrophobic amino acid side chains attached to the OTS surface with the hydrophilic side chains extending into the bulk solution. Since lysozyme is more stable at pH 7 than at pH 4, the difference in initial adsorption is dominated by the greater relative stability of lysozyme to denaturation at the higher pH. A change of pH from 7 to 4 reduces the stability of the protein to unfolding and results in more adsorption than when the pH is changed in the opposite direction. Solution pH also affects the net charges within the hydrophilic tail region and the structural distribution of the tail region was found to vary with pH. Copyright 1998 Academic Press.

Journal Article↗
PubMed2000

Effect of a Dynamic Stern Layer on the Sedimentation Velocity and Potential in a Dilute Suspension of Colloidal Particles.

In this paper the theory of the sedimentation velocity and potential (gradient) in a dilute suspension of charged spherical colloidal particles developed by Ohshima et al. (H. Ohshima, T. W. Healy, L. R. White, and R. W. O'Brien, J. Chem. Soc., Faraday Trans. 2, 80, 1299 (1984)) has been modified to include the presence of a dynamic Stern layer on the particle surfaces. The starting point has been the theory that Mangelsdorf and White (C. S. Mangelsdorf, and L. R. White, J. Chem. Soc., Faraday Trans. 86, 2859 (1990)) developed to calculate the electrophoretic mobility of a colloidal particle allowing for the lateral motion of ions in the inner region of the double layer (dynamic Stern layer). The effects of varying the different Stern layer parameters on the sedimentation velocity and potential are discussed and compared to the case when a Stern layer is absent. The influence of electrolyte concentration and zeta potential of the particles is also analyzed. The results show that regardless of the chosen set of Stern layer and solution parameters, the presence of a dynamic Stern layer causes the sedimentation velocity to increase and the sedimentation potential to decrease, in comparison with the standard case (no Stern layer present). These changes are almost negligible when sedimentation velocity is concerned, but they are very important when it comes to the sedimentation potential. A justification for this fact can be given in terms of an Onsager reciprocal relation, connecting the magnitudes of the sedimentation potential and the electrophoretic mobility. As previously reported, the presence of a dynamic Stern layer exerts a great influence on the electrophoretic mobility of a colloidal particle, and by means of the Onsager relation, the same is confirmed to occur when the sedimentation potential is concerned. Copyright 2000 Academic Press.

Journal Article↗
PubMed2002

Electrokinetics of concentrated suspensions of spherical colloidal particles with surface conductance, arbitrary zeta potential, and double-layer thickness in static electric fields.

In this paper the electrophoretic mobility and the electrical conductivity of concentrated suspensions of spherical colloidal particles have been numerically studied under arbitrary conditions including zeta potential, particle volume fraction, double-layer thickness (overlapping of double layers is allowed), surface conductance by a dynamic Stern layer model (DSL), and ionic properties of the solution. We present an extensive set of numerical data of both the electrophoretic mobility and the electrical conductivity versus zeta potential and particle volume fraction, for different electrolyte concentrations. The treatment is based on the use of a cell model to account for hydrodynamic and electrical interactions between particles. Other theoretical approaches have also been considered for comparison. Furthermore, the study includes the possibility of adsorption and lateral motion of ions in the inner region of the double layers (DSL model), according to the theory developed by C. S. Mangelsdorf and L. R. White (J. Chem. Soc. Faraday Trans.86, 2859 (1990)). The results show that the correct limiting cases of low zeta potentials and thin double layers for dilute suspensions are fulfilled by our conductivity formula. Moreover, the presence of a DSL causes very important changes, even dramatic, on the values of both the electrophoretic mobility and the electrical conductivity for a great range of volume fractions and zeta potentials, specially when double layers of adjacent cells overlap, in comparison with the standard case (no Stern layer present). It can be concluded that in general the presence of a dynamic Stern layer causes the electrophoretic mobility to decrease and the electrical conductivity to increase in comparison with the standard case for every volume fraction, zeta potential, and double-layer thickness.

Journal Article↗
PubMed2002

Rheology of PVC plastisol--VI: criteria for yielding and fracture of an immobilized layer.

PVC plastisol exhibits pseudo-plastic flow in steady shear; that is, viscosity decreases with the increasing shear rate. At higher shear rates viscosity reaches a minimum and then increases, i.e., dilatant behavior. Previously, pseudo-plastic behavior was explained by a mechanism in which the suspended particles partition into an immobilized layer and a mobile phase. The development of the immobilized layer with the increase in shear rate was shown to quantitatively account for pseudo-plastic behavior. In higher shear rates dilatation of the immobilized layer was shown to be the cause of dilatacy. At even higher shear rates the immobilized layer fractures. In this paper the viscosity minimum was interpreted as the yielding of the immobilized layer. Subsequently, data in the literature were analyzed to find criteria for the yielding and fracture of the immobilized layer. Yielding was found to obey Coulomb's criterion, from which the coefficient of friction and the cohesive strength of the immobilized layer were evaluated. These properties were controlled by the nature of particle assembly in the immobilized layer and the plasticizer type had only a minor effect. The value of the coefficient of friction was on the lower side and within the range of values found in the literature for other materials. There were two modes of fracture of the immobilized layer, one with low strength, low strain to break, and the other with high strength, high strain to break. The former is analogous to the brittle fracture of solids and the latter ductile failure. The strength of brittle fracture was somewhat higher than cohesive strength, which was evaluated from yielding data. This is akin to Griffith's criterion for brittle fracture of a solid. Ductile failure occurred when the shear stress exceeded normal stress.

Journal Article↗
PubMed1993

Double-layer prostheses for repair of abdominal wall defects in a rabbit model.

The primary objective of this study was to compare the effectiveness of expanded polytetrafluoroethylene (e-PTFE) and polypropylene (PPM) in a rabbit abdominal wall defect model using a double-layer repair technique. Fifty-four New Zealand white rabbits had full thickness resection of a 5 x 5-cm segment of the mid portion of at the abdominal wall. The defect was repaired with two 6 x 6-cm layers of prosthesis in an underlay/overlay fashion incorporating a 1-cm edge of abdominal wall and grouped accordingly. Group I: inner and outer layer of PPM; Group II: inner layer e-PTFE, outer layer PPM; Group III: inner and outer layer e-PTFE. Animals from each group were sacrificed at 3, 6, and 12 months. The abdominal wall was assessed for herniations and bowel adhesions to the inner layer of the prosthesis. Adhesions were graded according to an adhesion scoring system (grade 0-3). No herniations were observed. Intestinal adhesions to PPM were quite dense at 3 months with a mean adhesion score of 2.5; at 12 months, adhesions were more dense and extensive, having a mean score of 3. Adhesions to e-PTFE were thin and filmy with a mean score of 0.4 at 3 months and 0.6 at 12 months (P < 0.001, 95% CI for each time period). Histologically, PPM fibers were consistently surrounded by dense fibrous connective tissue; foreign body giant cells were present. A thin connective tissue capsule covered the inner e-PTFE layer; its pores were progressively infiltrated by fibroblasts. The double-layer technique prevented hernias in all groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Abdominal Muscles↗
PubMed1998

On the concept of leaf boundary layer resistance for forced convection

The definition of leaf boundary layer resistance is reconsidered in respect of the three-dimensional diffusion-controlled mass transport region just above the leaf surface. Due to the existence of this superstomatal air layer, the conventional convective boundary layer is not in direct contact with the surface. Thus, in terms of plant physiology, the diffusive "end correction" to the stomatal resistance should be included in the boundary layer resistance. This is true for laminar as well as turbulent flows. When the surface mole fraction of an exchanged gas is estimated using the boundary layer resistance ignoring the diffusive term may lead to a noticeable error. The self-consistent approach is used to clarify the problems of the boundary layer formation and stomatal interference. If the correction is taken into account, the boundary layer resistance becomes dependent also on stomatal shape and distribution on the leaf. The traditional semiempirical formula corrected by the superstomatal diffusion is applied in numerical calculations. In estimates of the water vapour mole fraction on the surface of a transpiring leaf the relative error ranges from insignificant (quiescent air, large leaf and large stomatal pores) to 20 % (low humidity, strong wind, small leaf and small elliptic pores). The boundary layer resistance can decrease by a factor of 3 when the semiaxis lengths of the stomata increase from 1 and 0.5 &mgr;m to 10 and 5 &mgr;m. The effective thickness of the superstomatal air layer is maximally several millimetres (small stomatal surface concentration and small pores). Copyright 1998 Academic Press Limited

Journal Article↗
PubMed2001

Modelled influences of non-exchanging trichomes on leaf boundary layers and gas exchange.

The two main resistances in the exchange of gases between plants and the atmosphere are stomatal and boundary layer resistances. We modeled boundary layer dynamics over glabrous and pubescent leaves (assuming non-exchanging trichomes) with leaf lengths varying from 0.01 to 0.2 m, and windspeeds of 0.1-5.0 m x s(-1). Results from theoretical and semi-empirical formulae were compared. As expected, boundary layer thickness decreased with decreasing leaf length and increasing windspeed. The presence of trichomes increased leaf surface roughness, resulting in lowered Reynolds numbers at which the boundary layer became turbulent. This effect is especially important at low windspeeds and over small leaves, where the Reynolds number over glabrous surfaces would be low. We derived a new simple dimensionless number, the trip factor, to distinguish field conditions that would lead to a turbulent boundary layer based on the influence of trichomes. Because modeled rates of CO2 and H2O(v) exchange over turbulent boundary layers are one or more orders of magnitude faster than over laminar boundary layers, a turbulent boundary layer may lead to increased carbon uptake by plants. The biological trade-off is potentially increased transpirational water loss. However, in understory habitats characterized by low windspeeds, even a few trichomes may increase turbulence in the boundary layer, thus facilitating photosynthetic gas exchange. Preliminary field data show that critical trip factors are exceeded for several plant species, both in understory and open habitats.

Carbon Dioxide↗
PubMed1986

The innermost cell layer of the outer root sheath in anagen hair follicle: light and electron microscopic study.

To investigate cell differentiation in the outer root sheath (ORS) of the human anagen hair follicle, scalp skin specimens from individuals with normal hair were examined using light and electron microscopes. In the bulbar portion, the ORS was composed of two cell layers. The cells in the outer layer gradually increased in number upwards and finally underwent so-called trichilemmal keratinization, which proceeded toward the hair canal. On the other hand, the inner cells in the bulb formed a single cell layer along the outside of Henle's layer during cell differentiation; this unique layer was referred to as the innermost cell (IMC) layer of the ORS. With the use of hematoxylin and eosin stain, at the suprabulbar portion, where Henle's cells were keratinizing, an eosinophilic substance was deposited in the inner (Henle's) side of the IMC cytoplasm. The IMCs gradually became entirely eosinophilic and often produced keratohyaline granules. Ultrastructurally, the IMCs of the ORS showed an oblong shape forming a regularly arranged single-cell layer along the keratinizing Henle's layer and accumulated tonofilaments in the cytoplasm. They produced a few small electron-dense keratohyaline granules and were keratinized at the level at which Henle's layer still preserved its cell structure. From these findings, it is suggested that there are two types of keratinization of the ORS: trichilemmal keratinization and IMC keratinization.

Cell Differentiation↗
PubMed1986

Structural and chemical characterization of S-layers of selected strains of Bacillus stearothermophilus and Desulfotomaculum nigrificans.

The structures, amino acid- and neutral sugar compositions of the crystalline surface layers (S-layers) of four selected strains each of Bacillus stearothermophilus and Desulfotomaculum nigrificans were compared. Among the four strains of each species a remarkable diversity in the molecular weights of the S-layer subunits and in the geometry and constants of the S-layer lattices was apparent. The crystalline arrays included hexagonal (p6), square (p4) and oblique (p2) lattices. In vitro self-assembly of isolated S-layer subunits (or S-layer fragments) led to the formation of flat sheets or open-ended cylindrical assembly products. The amino acid composition of the S-layers exhibited great similarities and was predominantly acidic. With the exception of the S-layers of two strains of B. stearothermophilus (where only traces of neutral sugars could be detected), all other S-layer proteins seemed to be glycosylated. Among these strains significant differences in the amount and composition of the glycan portions were found. Based on this diversity interesting questions may be asked about the biological significance of the carbohydrate units of glycoproteins in prokaryotic organisms.

Amino Acids↗
PubMed1992

A separating method for quantifying mucus glycoprotein localized in the different layer of rat gastric mucosa.

A method was devised for separating rat gastric mucosa into three layers each containing a different mucin species. The mucus gel (first layer) was removed by stirring the gastric mucosa in a solution of phosphate-buffered saline containing 2% N-acetylcysteine. The surface mucosa (second layer), rich in surface mucus cells, was then separated from the deep mucosa (third layer) containing mucus neck cells, by scraping with forceps. The effectiveness of this method was confirmed by light microscopical observation after GOCTS-PCS (dual staining by the galactose oxidase-cold thionin Schiff method and paradoxical concanavalin A method) and AB-PAS staining (dual staining with alcian blue and the periodic acid Schiff method). The fixed specimen of scraped mucus and cell debris was rich in AB-PAS and GOCTS positive mucus, but was hardly stained by PCS, indicating mucus derived from surface mucus cells to have been efficiently recovered from this preparation. The residual mucosa could be stained by PCS but hardly at all by AB-PAS or GOCTS. The lyophilized powder specimens obtained from the three different layers of rat gastric mucosa were used to extract and quantify mucus glycoprotein (mucin). This was done to examine changes in mucin content in the three layers of gastric mucosa one hour following the oral administration of 20% ethanol or 0.35 N hydrochloric acid, both mild irritants. Mucin content was noted to significantly increase in the first layer but hardly at all in the second layer. In the third layer, it decreased significantly by 0.35 N hydrochloric acid, but changed only slightly by 20% ethanol administration.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗