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PubMed1991

The egg-shell of Drosophila melanogaster. VI, Structural analysis of the wax layer in laid eggs.

Utilizing freeze-fracturing conventional electron microscopy and scanning electron microscopy methods, a wax layer was identified, sealing the oocyte of Drosophila melanogaster. In mature egg-shells wax forms a hydrophobic layer surrounding the oocyte and lying between, and in very close contact with the vitelline membrane (interiorly) and the crystalline intermediate chorionic layer (exteriorly). In cross-fractured views it is less than 50 A thick whereas in longitudinal fracturing it reveals smooth fracture faces of a multilayered material in the form of hydrophobic areas or plaques (0.5-1 microns in diameter) which are partially overlapping and highly compressed between the vitelline membrane and the innermost chorionic layer. The evidence for this layer being a wax are the facts that a) it is not preserved in conventional fat-extracting electron microscopy methods, b) it directs laterally the fracture planes during freeze-fracturing and reveals smooth fracture faces. Analysis of the structural features of wax in mature egg-shell in various species of Drosophilidae have shown that the wax layer exhibits indistinguishable (among the species) hydrophobic plaques, which have the same size and thickness with Drosophila melanogaster. These data provide structural evidence explaining the physiological resistance of the insect eggs studied, against water loss or water uptake, whenever they are laid on substrates with extreme environmental conditions. In addition, the data demonstrate how an extracellular substance can be organized to perform that function.

Animals↗
PubMed1982

Proteins of the outer layer of the vitelline membrane of hen's eggs.

A study has been made of the proteins in the vitelline membrane of hen's eggs before and after mechanical separation into the inner and outer layers. The membranes were dissolved in detergent (sodium dodecyl sulphate) and chromatographic fractions were examined by gel electrophoresis. The separated inner and outer layers were compared by gel electrophoresis. The outer layer contained (i) enzymically active lysozyme (EC 3.2.1.17) (about 60% dry weight), (ii) an insoluble ovomucin complex and (iii) a new protein, VMOI (vitelline membrane outer I). These account for most of the protein. In addition, some minor constituents were detected by gel electrophoresis but were not isolated. Except for ovomucin, the constituents of the outer layer could be dissolved from the membrane at high ionic strength (greater than 0.5 M sodium chloride), resulting in a loss of its structure. On lowering the ionic strength the soluble proteins recombined with the membrane, partially regenerating the original structure. Ovomucin appears to form the skeleton of the outer layer, but the salt-soluble proteins, especially lysozyme, are responsible for its integrity. The function of the newly-recognized protein (VMOI) is not known. Its molecular weight is 17,500 according to gel electrophoresis in detergent and it contains no methionine. The inner layer consists largely of the proteins GPI, GPII and GPIII isolated by Kido et al. (Kido, S., Janado, M. and Nunoura, H. (1975) J. Biochem. 78, 261-268) from the whole membrane.

Animals↗
PubMed1993

Postnatal ontogeny of GABAA and benzodiazepine receptors in individual layers of rat visual cortex and the effect of visual deprivation.

The postnatal development of GABAA and benzodiazepine receptors was studied in the rat visual cortex with [3H]muscimol and [3H]flunitrazepam, respectively. The studies were performed by quantitative receptor autoradiography in the different cortical layers from normally raised rats and from monocularly deprived animals. In all cortical layers of normal rats there was a sharp (approx 3-fold) increase in [3H]muscimol binding from days 1 to 10. Between postnatal days 10 and 20 there was a considerable decrease in GABAA receptors; thereafter this decline continued until adulthood but at a more moderate rate. In all cortical layers, [3H]flunitrazepam binding increased sharply, also from day 1 to 10; at this age the adult values were practically attained. Monocular deprivation, by unilateral eyelid suture at the age of 11 days onwards lead to an increase in [3H]muscimol binding in cortical layers IV-VI. This increase in GABAA receptors was detectable at the age of 25 days onwards. In contrast, monocular deprivation did not alter the density of benzodiazepine receptors in any of the cortical layers, and this lack of effects was regardless of the duration of visual deprivation. The present results suggest that the development of GABAA receptors in the deeper cortical layers, but not that of benzodiazepine receptors, is seemingly regulated by the visual experience at a critical period of brain maturation.

Animals↗
PubMed1995

Organization of the outer layers of the cell envelope of Corynebacterium glutamicum: a combined freeze-etch electron microscopy and biochemical study.

The cell surface of Corynebacterium glutamicum grown on solid medium was totally covered with a highly ordered, hexagonal surface layer. Also, freeze-fracture revealed two fracture surfaces which were totally covered with ordered arrays displaying an hexagonal arrangement and the same unit cell dimension as the surface layer. The ordered arrays on the concave fracture surface, closest to the cell surface, were due to the presence of particles while those on the convex fracture surface were their imprints. The same cells grown on liquid medium displayed a cell surface and fracture surfaces only partially covered with ordered arrays. In this case, the ordered regions had the same relative position on the cell surface and on the fracture surfaces. All ordered arrays were totally absent in a mutant for cspB, the gene encoding PS2, one of the two major cell wall proteins. Treatment of the cells with proteinase K caused the gradual alteration of PS2 into a slightly lower molecular mass form. This was accompanied by a concomitant disappearance of the ordered fracture surfaces followed by the detachment of the ordered surface layer from the cell as large ordered patches displaying the same lattice symmetry and dimensions as those of the surface layer. The ordered patches were isolated. They contained the totality of PS2 initially associated with the cell. We conclude that the highly ordered surface layer of the intact cell was composed of PS2 interacting strongly with some cell wall material leading to its organization. This organized cell wall material produced fracture surfaces. We show that in the absence of intact PS2 protein on the cell wall, the same cell wall material was not organized and formed a structureless smooth layer.

Bacterial Proteins↗
PubMed1986

Phased array ultrasound imaging through planar tissue layers.

Conventional ultrasound imaging devices are designed based on the assumption of a homogeneous tissue medium of constant acoustic velocity = 1540 m/sec. However, the body consists of tissue layers of varying thicknesses and velocities which range from 1470 m/sec in fat to 3200 m/sec in skull bone. Refraction effects from these layers degrade ultrasound image quality. In this paper, pulse-echo ultrasound imaging is modeled as imaging an organ of interest through an intervening planar tissue layer, such as liver through fat in the abdomen or brain through skull bone in the adult head. Refraction effects from planar tissue layer interfaces are analyzed using Snell's law and measured using phantoms. We also introduce an on-line phased array correction technique based on planar tissue layers to restore ultrasound image quality. We conclude that fat/organ planar interfaces do not degrade image quality significantly. However, refraction effects at a skull/brain planar interface degrades resolution and target acquisition and introduces geometric distortion. Our plane layer phased array correction technique significantly improves image quality in phantoms through lucite aberrators and improves adult cephalic ultrasound image quality when used through the top of the adult skull. The correction technique is robust even in the presence of inaccurate estimates of skull thickness.

Adipose Tissue↗
PubMed1985

A projection from the deep layers of the entorhinal area to the hippocampal formation in the rat brain.

The methods of retrograde fluorescent tracing and anterograde transport of the lectin Phaseolus vulgaris leucoagglutinin (PHA-L) were used to demonstrate the existence of projections from layers IV and VI of the entorhinal area to the hippocampal formation in the rat brain. These two layers of the medial and lateral entorhinal area innervate the molecular layer of Ammon's horn and the area dentata. In the area dentata the projection from layer IV follows that of the perforant path, while that from layer VI innervates the outer two-thirds of the molecular layer, the subgranular zone and the deep part of the hilus of the area dentata.

Amidines↗
PubMed1982

Anterograde tracer and field potential analysis of the neocortical layer I projection from nucleus ventralis medialis of the thalamus in cat.

The projection of the ventromedial nucleus of the thalamus to the neocortex was studied in cat by means of anterograde and retrograde transport of horseradish peroxidase, by the depth profile of evoked thalamocortical field potentials, and by superfusion of the cortex with manganese to block transmitter release. Horseradish peroxidase injected into the ventromedial nucleus was anterogradely transported to the outer third of layer I in the neocortex, extending from the depth of the cruciate sulcus anterior to the olfactory bulb and tract. The region of projection from the ventromedial nucleus extended mediolaterally from the medial wall of the proreus gyrus to the ventral tip of the coronal gyrus. Horseradish peroxidase injections or applications in these areas of the neocortex resulted in the retrograde labeling of neurons in the ventromedial nucleus. Injections in many other cortical areas did not result in labeled neurons in this nucleus. Stimulation of the ventromedial nucleus with single pulses elicited surface-negative waves in the medial part of the precruciate region that had superficial isoelectric points. Superfusion of the precruciate area with manganese resulted in the suppression of the ventromedial-evoked wave, whereas control extracellular waves in deeper layers were unaffected. An additional additional finding was that horseradish peroxidase injections in the ventromedial nucleus led to a dense reciprocal retrograde labeling of neurons in layer VI of that part of the cortex to which the ventromedial nucleus projects. We conclude that, in cat, (1) the ventromedial nucleus projects to layer I of the cerebral cortex anterior to the cruciate sulcus and receives a dense reciprocal projection from layer VI; (2) stimulation of neurons in the ventromedial nucleus causes depolarization of structures in layer I and these neurons are responsible for recruiting responses in the anterior cortex.

Action Potentials↗
PubMed1986

Complementary and non-matching afferent compartments in the cat's superior colliculus: innervation of the acetylcholinesterase-poor domain of the intermediate gray layer.

Three tectal afferent-fiber systems were experimentally labeled in the cat to learn how their distributions within the superior colliculus were related to the prominent compartments of high acetylcholinesterase activity found in the intermediate gray layer. Presumptive somatic sensory afferents were labeled by injections of horseradish peroxidase-wheatgerm agglutinin conjugate placed at the bulbospinal junction and in the ventral anterior ectosylvian cortex corresponding to somatic sensory area SIV. Vision-related afferents were labeled by injections of the same tracer substance into the lateral suprasylvian visual area. In each animal, a single type of injection was made and a detailed study was carried out to compare the patterns of anterograde labeling and acetylcholinesterase staining in serially adjoining sections through the superior colliculus. Fibers labeled by the three types of injection were distributed in clusters that resembled the acetylcholinesterase-positive patches in the intermediate gray layer. In no case, however, were the afferent-fiber clusters in register with the histochemically defined patches. Instead, the innervations derived from the bulbospinal junction, anterior estosylvian sulcus and lateral suprasylvian visual area all formed patchworks within the acetylcholinesterase-poor domain of the intermediate gray layer. In some instances, the afferent-fiber clusters and enzyme-positive patches appeared to have complementary distributions. In other instances, the afferent-fiber clusters seemed to be arranged in the acetylcholinesterase-poor parts of the intermediate layer in a fashion independent of, but not significantly overlapping, the acetylcholinesterase-positive patches. Not all of the space between the acetylcholinesterase-positive patches was taken up by any one of the afferent-fiber systems labeled. The complementary and non-matching distribution of these afferent systems in relation to the acetylcholinesterase-rich patches of the intermediate gray layer stands in contrast to the spatial registration of two other tectal afferent systems with the zones of high acetylcholinesterase activity. Both nigrotectal and frontotectal afferents converge on the acetylcholinesterase-positive patches. We conclude that afferent systems projecting to the intermediate gray layer can be divided into at least two groups: those innervating the acetylcholinesterase-rich compartments and those avoiding them.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholinesterase↗
PubMed1990

Segregation and heterogeneity of thalamic cell populations projecting to superficial layers of posterior parietal cortex: a retrograde tracer study in cat and monkey.

The thalamic neurons projecting to the superficial layers of areas 5 and 7 in the cat, and area 5 in the monkey, were investigated by using superficial deposits of either horseradish peroxidase or Fast Blue in one hemisphere. In the contralateral hemisphere injections of the same tracer involving the full cortical depth were made in homotopical locations, and the distribution and soma size of retrogradely labeled thalamocortical neurons in each side of the thalamus were compared. It was found that, in the cat, labeled neurons in the lateral posterior pulvinar complex, and in paralaminar regions of the ventrolateral complex, were fewer in number and smaller in size in cases of superficial deposits than in cases of deep injection. In more lateral portions of the ventrolateral complex, however, there were no size differences. In the monkey, similar differences in number and size appeared in the caudal division of the ventrolateral complex and in the lateral posterior and pulvinar nuclei, whereas no such differences were found for neurons labeled in the oral and medial divisions of the ventrolateral complex, and in the ventral posteroinferior nucleus. In all cases the intralaminar and midline nuclei exhibited retrogradely labeled neurons only when deep layers were injected. These and previous findings point to the existence of a widely distributed layer I-projecting system of neurons which, in most nuclei, are interspersed among neurons projecting mainly to middle or deep layers. In some nuclei, however, as is the case with the ventromedial nucleus proper, layer I-projecting system neurons would make up the whole nucleus. The cell groups located in a paralaminar position, which would be but a part of this system, could provide through their projections to layer I in the posterior parietal and frontal cortical regions a final path for recruiting responses and spontaneous spindling activities.

Amidines↗
PubMed2005

The effect of single- or multiple-layered garments on interface pressure measured at the backpack-shoulder interface.

Experiments were aimed at determining whether interface pressure measurements are a true reflection of skin contact pressure when made over different layers of clothing. Interface pressures were recorded at the skin interface and above clothing layers, with 11 participants wearing 10 different clothing combinations. The clothing used was standard issue British military clothing, including body armour (without ballistic plate). Participants wore both single and multiple clothing layers whilst walking on a treadmill carrying a loaded backpack (23.5 kg, British military issue). Results showed no significant differences (p=>0.05) in pressure between different clothing layers either singly, or worn in multiple. In conclusion, the soldier (or leisure user) will gain no or very little relief from applied pressure by wearing garments, even in layers, when carrying a backpack. Also, the findings suggest that interface pressure may adequately be assessed using a sensor placed above the clothing layer(s) rather than at skin surface.

Adult↗
PubMed2005

Immobilization of hemoglobin at the galleries of layered niobate HCa2Nb3O10.

Hemoglobin (Hb) was intercalated at the galleries of layered niobate HCa(2)Nb(3)O(10) (HCNO). Two different kinds of layered phases of Hb-CNO composites Hb-CNO-1 and Hb-CNO-2 were obtained with the interlayer distances of 7.2 and 10.3nm in correspondence with the monolayer and bilayer arrangements of proteins between the niobate layers, respectively, based on the powder XRD pattern, HRTEM, UV-vis spectra and CHN analyses. FTIR spectra of Hb-CNO composites show that amide I and amide II bands were actually the same as those of the native Hb, which indicates that there is almost no structural change after immobilization. Michaelis-Menten model methods were used to study the peroxidatic activity of the reaction of 2-methoxyphenol and H(2)O(2) for the entrapped Hb in the galleries of HCNO. Compared to that of free Hb, the kinetic parameters of Hb-CNO k(cat), K(M) and k(cat)/K(M) were affected by the immobilization process. The immobilized Hb showed a higher relative activity than that of free Hb after incubated in phosphate buffer (pH = 7) at 80 degrees C for a period of time. The environments between the layers of HCNO are hydrophilic which will bind water tightly and help to stabilize the 'essential water' layer around the protein. So, immobilization of Hb between the layers of HCNO enhanced the activity of Hb in water-DMSO mixture.

Adsorption↗
PubMed2006

The role of independently variable grafting density and layer thickness of polymer nanolayers on peptide adsorption and cell adhesion.

This contribution demonstrates a simple and reproducible method for fabricating surface-tethered polymer brushes that vary in grafting density and layer thickness for peptide adsorption and cell-adhesion studies. Surface-initiated atom transfer radical polymerization was used together with thiol self-assembly to generate these nanothin polymer brush layers of poly((polyethylene glycol) methacrylate). A kinetic study was done to measure the layer thickness growth rate at room temperature from flat gold substrates presenting different polymerization initiator molecule surface densities. The polymer brush layers transition from mushroom to brush regimes with increasing grafting density. A crossover density of 0.038+/-0.005 chains/nm(2) was determined for the PPEGMA polymer brushes. The results described in this paper show that layer properties such as wettability and dry layer thickness depend strongly on initiator surface density. Ultimately, the adsorbed concentration of an RGD-containing synthetic peptide Gly-Arg-Gly-Asp-Ser and the adhesion and spreading of cells were correlated with surface properties, which continues to be a major research theme in biomedical and biomaterials research.

3T3 Cells↗
PubMed2005

Dorsal raphe nucleus stimulation modulates the response of layers IV and V barrel cortical neurons in rat.

The effect of dorsal raphe nucleus (DRN) electrical stimulation on response properties of layers IV and V barrel cortical neurons was studied. To assess the receptive field characteristics of cortical neurons, responses of neurons were recorded following the displacement of principal and adjacent whiskers individually or in a condition test paradigm. Then neuronal responses to the displacement of whiskers were analyzed following DRN stimulation at 0, 50, 100, 200 and 400 ms inter-stimulation intervals. Considering On responses, DRN stimulation suppressed the response magnitude of layer V neurons to principal whisker deflection, while it slightly increased that of layer IV neurons (not statistically significant). The response latency of layer IV neurons increased when DRN was stimulated 200 or 400 ms before principal whisker deflection, while the response latency of layer V was not changed. DRN stimulation had no effect on either magnitude or latency of neuronal response to the adjacent whisker deflections. We observed a decrease in the inhibitory effect of the adjacent whisker deflection on the magnitude of neuronal response to the principal whisker deflection in layer IV when DRN was stimulated 200 ms before the principal whisker deflection. Off responses did not show any significant effect of DRN stimulation. Our results suggest a modulating role for DRN in processing of the incoming information into barrel cortex. This effect might be location dependent.

Action Potentials↗
PubMed2006

Short-term synaptic plasticity in layer II/III of the rat anterior cingulate cortex.

Recent in vivo electrophysiological studies in our laboratory demonstrated medial thalamus (MT) induced short-term facilitation in the middle layers of the anterior cingulate cortex (ACC). The aim of the present study was to investigate different forms of short-term plasticity (STP) in layer II/III of the ACC in an in vitro slice preparation. Extracellular field potentials in layer II/III consisting of an early component (fAP) and a late component (fPSP) were activated by electrical stimulation of the deep layers. The fPSP and intracellularly recorded excitatory post-synaptic potential (EPSP) could be facilitated by paired-pulse stimulation at a low frequency (0.033Hz, pulse interval 20-400ms). An initial facilitation and subsequent depression were obtained when high frequency (12.5, 25 and 50Hz) tetanus stimulations were applied to the ACC slice. A post-tetanic augmentation 30s in duration was also observed. The effects of tetanic stimulation were altered in the presence of an increased or a decreased calcium concentration. Application of omega-conotoxin GVIA (CTX) in normal calcium concentration conditions decreased overall responses during tetanic stimulation similar to reducing calcium exposure. However CTX application did not increase paired-pulse facilitation (PPF) as is seen under low calcium conditions. These results indicate that calcium is involved in the formation of certain features of STP in layer II/III of the ACC and that N-type calcium channels contribute to some, but not all, components of these plastic changes. Two-site electrical stimulation testing showed that two separate presynaptic inputs can produce short-term facilitation. Our findings implicate a post-synaptic mechanism in STP in layer II/III of the ACC.

Animals↗
PubMed2006

Heat production and lipid metabolism in broiler and layer chickens during embryonic development.

We compared heat production (HP) and lipid metabolism in broiler and layer chickens (Gallus gallus) during embryonic development. To investigate HP and respiratory quotient (RQ), oxygen (O2) consumption and carbon dioxide (CO2) production were measured using an open-circuit calorimeter system. HP consistently had a tendency (P = 0.06) to be lower in broilers than in layers during embryonic development, and HP gradually decreased with developmental stage in both strains. RQ values of both strains were approximately 0.7 at every embryonic stage investigated. These results suggest that chicken embryos mainly use lipid for energy, and the RQ was significantly lower in broilers than in layers during embryonic development. Consumption of the yolk sac as a lipid source was faster in broilers than in layers. Plasma D-3-hydroxybutyrate (D3HB) and glycerol concentrations, associated with fatty acid oxidation, were lower in broiler than layer embryos. These results demonstrate that HP and lipid metabolism are different between the strains during embryonic development, and may be one factor for the growth difference between broiler and layer embryos.

3-Hydroxybutyric Acid↗
PubMed2005

ICV CRF and isolation stress differentially enhance plasma corticosterone concentrations in layer- and meat-type neonatal chicks.

The present study compared the plasma corticosterone concentrations between meat- and layer-type neonatal chicks (Gallus gallus) (1) exposed to isolation-induced stress or (2) injected intracerebroventricularly (ICV) with corticotropin-releasing factor (CRF). Both types of neonatal chicks housed in groups were individually introduced to an open field arena and locomotion and distress-induced vocalizations were monitored for 10 min. The responses of the two strains were remarkably different, with meat-type chicks being less active than layer-type chicks. Distress-induced vocalizations were drastically decreased over time in meat-type chicks while they remained high in layer-type chicks throughout the test. Plasma corticosterone concentrations measured at the end of the test were significantly higher in layer-type chicks than in meat-type ones. Plasma corticosterone concentrations measured 10 min after the ICV injection of CRF were significantly higher in layer- than meat-type chicks. These results indicate that meat-type chicks have either a greater capability to acclimatize to novel environments, or a blunted HPA axis compared with layer-type chicks.

Animals↗
PubMed2006

Treatment of phenolic wastewater by horseradish peroxidase immobilized by bioaffinity layering.

Horseradish peroxidase was immobilized by bioaffinity layering and used for the treatment of wastewater containing p-chlorophenol. For this purpose, lectin Concanavalin A was bound to Sephadex beads. The glycoenzyme peroxidase was layered upon this Con A layer. Subsequently, alternate layers of the enzyme and Con A were applied. The most efficient design consisted of three layers of Con A and peroxidase each. This immobilized enzyme preparation retained 80% of the activity of the free peroxidase used for immobilization. PEG at the concentration of 0.1 mg ml(-1) was found to prevent enzyme inactivation by the products, although it increased the process time. Thus 60 U ml(-1) of enzyme completely converted the p-chlorophenol (into products) in 4 min in the absence of PEG. On the other hand, only 0.05 U ml(-1) of enzyme was required for this purpose in the presence of PEG but the process required 60 min. Peroxidase converts phenol molecules into free radicals. These free radicals then polymerize and get precipitated. As a further means of minimizing exposure of the enzyme to free radicals and enhancing the reusability, it was decided to remove the enzyme from reaction medium after 10 min. With this strategy, the bioaffinity layered peroxidase preparation could be reused five times without any loss of activity.

Chlorophenols↗
PubMed2006

Composite interfacial layers containing micro-size and nano-size particles.

Surface layers of micro- and nanoparticles at fluid/liquid interfaces in absence and presence of surfactants are of large importance in the process of re-discovering Pickering systems, i.e. emulsions and foams stabilized by particles. The surface pressure/area isotherms of such layers can provide information about the properties of the used particles (dimensions, interfacial contact angles), the structure of interfacial layers, the interactions between the particles as well as about relaxation processes within the layers. For a correct description of Pi-A isotherms of composite surface layers containing particles the significant difference in size of these particles to that of solvent and surfactant molecules should be taken into account. Corresponding equations can be derived on the basis of the two-dimensional solution theory. The gained equations provide satisfactory agreement with experimental data and predict realistic values for the area of particles at the interface. Also equations of state and of the dilational elasticity for composite surface layers containing particles can be obtained in the framework of the presented methodology.

Adsorption↗