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PubMed1996

Erythrocyte flow and elasticity of microvessels evaluated by marginal cell-free layer and flow resistance.

Flow dynamics of human erythrocytes was compared in elastic (E) and hardened (H) microvessels with inner diameters of 10-40 microns. The thickness of the marginal cell-free layer and the overall flow resistance were measured with a vascular bed isolated from rabbit mesentery (E vascular bed) as well as with a 4% paraformaldehyde-fixed bed (H vascular bed). 1) In both E and H microvessels, the thickness of the cell-free layer increased with increasing inner diameter of the microvessels and with decreasing hematocrit accompanied by an overall decrease in the flow resistance. The hematocrit-dependent change of the cell-free layer thickness was greater in the E microvessels than in the H microvessels. The flow resistance was always greater in the H vascular beds than in the E vascular beds. 2) With decreasing erythrocyte deformability induced by treatment with 2 mM diazenedicar-boxylic acid bis(N,N-dimethylamide), the thickness of the cell-free layer decreased at a low hematocrit in the E microvessels and at a high hematocrit in the H microvessels, although the flow resistance was increased in both vascular beds. 3) Dextran of 70,400 average molecular weight accelerated the formation of the cell-free layer by inducing erythrocyte aggregation. A drastic increase in the cell-free layer thickness at 2-4 g/dl of dextran in the E microvessels and at 1-2 g/dl of dextran in the H microvessels was accompanied by a significantly lower increase in the flow resistance. This study concludes that the elasticity of microvessels may play an important role for reducing the overall flow resistance of a vascular bed, which is modulated by the marginal cell-free layer, itself a function of the rheological properties of the erythrocytes.

Animals↗

PubMed1993

Correlation of physiological subgroupings of nonpyramidal cells with parvalbumin- and calbindinD28k-immunoreactive neurons in layer V of rat frontal cortex.

1. To test the hypothesis that physiologically and morphologically different cortical nonpyramidal cells express different calcium-binding proteins, whole-cell current-clamp recording in vitro was combined with intracellular staining and double immunofluorescence in layer V of frontal cortex of rats 16-20 days old. 2. Nonpyramidal cells were first characterized as fast-spiking (FS) or low-threshold spike (LTS) cells, injected with biocytin, and subsequently stained immunohistochemically for parvalbumin and calbindinD28k. 3. FS cells were identified by input resistances < 350 M omega, spike width at half amplitude < 0.8 ms, and virtually no spike frequency adaptation of spike trains by depolarizing pulses. LTS cells were identified by input resistances > 350 M omega, spike width at half amplitude > 0.8 ms, and the discharge of low-threshold spikes from hyperpolarized potentials. Repetitive firing could be induced by a combination of stimulation-induced excitatory postsynaptic potentials with depolarization in FS cells. Repetitive firing was not observed in LTS cells under these conditions. 4. After biocytin injection of layer V cells characterized in this way, subsequent double immunostaining showed that all biocytin-labeled parvalbumin-immunoreactive cells (n = 18) belonged to the FS cells (FS-PV cells), whereas all biocytin-labeled calbindinD28k-immunoreactive cells (n = 10) belonged to the LTS cells (LTS-Calb cells). 5. FS-PV cells had smooth or sparsely spiny dendrites, whereas LTS-Calb cells had dendrites with a modest number of spines but fewer than pyramidal cells. FS-PV cells showed denser axonal branches near their somata and extended axons in a more horizontal direction. Some of them could be identified as basket cells by the presence of terminal boutons surrounding somata of other cells. LTS-Calb cells extended their main axons more vertically up to layer I. 6. Double immunofluorescent staining revealed that very few cells in layer V showed immunoreactivity for both calcium-binding proteins but that most cells immunoreactive for the calcium-binding proteins in layer V were also immunoreactive for gamma-aminobutyric acid. 7. These results suggest that GABAergic nonpyramidal cells in layer V of neocortex can be divided into two functional groups on the basis of different firing modes, axonal distributions, and calcium-binding protein immunoreactivity: 1) FS-PV cells show repetitive firing by synaptic activation, have axonal arborizations that are more dense near their somata and oriented horizontally, and the cells exhibit parvalbumin immunoreactivity and 2) LTS cells show low-threshold spikes, have more vertical axonal arborizations up to layer I, and exhibit calbindinD28K immunoreactivity.

Animals↗

PubMed1994

Differences in voltage-dependent sodium currents exhibited by superficial and deep layer neurons of guinea pig entorhinal cortex.

1. Sodium currents were studied using whole-cell voltage-clamp techniques in neurons acutely isolated from superficial (II/III) and deep (V/VI) layers of guinea pig entorhinal cortex. 2. Sodium currents were larger (peak amplitude) in superficial than in deep layer cells under the same conditions: -1939 +/- 780 (SD) pA (N = 6) versus -307 +/- 257 pA (N = 6). Specific membrane conductance was calculated to be 12.3 +/- 9.6 mS/cm2 for superficial layer cells and 1.4 +/- 0.9 mS/cm2 for deep layer cells. 3. Sodium currents could be activated in superficial layer cells from potentials as depolarized as -20 mV, whereas no significant currents could be activated in deep neurons from potentials more depolarized than about -50 mV. Using a protocol consisting of a 25-ms prepulse and a 20 ms test pulse, the inactivation curves for superficial layer cells were found to be shifted toward more depolarized potentials by an average of 15 mV (V50 = -59.8 +/- 3.8 mV compared with -75.7 +/- 12.0 mV for deep cells). This produced a region of overlap with the activation curves for superficial cells. 4. Over a range of about -50 to -20 mV in superficial layer cells, the region of overlap of the activation and inactivation curves, a sodium current could be activated, which did not fully inactivate during the test pulse (average peak amplitude: -89.5 +/- 48.7 pA; crossover voltage: -39.2 +/- 2.0 mV). Voltage steps to more depolarized potentials, outside the voltage "window", permitted complete inactivation of the sodium current.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2000

An excitatory GABAergic plexus in developing neocortical layer 1.

Layer 1 of the developing rodent somatosensory cortex contains a dense, transient GABAergic fiber plexus. Axons arising from the zona incerta (ZI) of the ventral thalamus contribute to this plexus, as do axons of intrinsic GABAergic cells of layer 1. The function of this early-appearing fiber plexus is not known, but these fibers are positioned to contact the apical dendrites of most postmigratory neurons. Here we show that electrical stimulation of layer 1 results in a GABA(A)-mediated postsynaptic current (PSC) in pyramidal neurons. Gramicidin perforated patch recording demonstrates that the GABAergic layer 1 synapse is excitatory and can trigger action potentials in cortical neurons. In contrast to electrical stimulation, activation of intrinsic layer 1 neurons with a glutamate agonist fails to produce PSCs in pyramidal cells. In addition, responses can be evoked by stimulation of layer 1 at relatively large distances from the recording site. These findings are consistent with a contribution of the widely projecting incertocortical pathway, the only described GABAergic projection to neonatal cortex. Recording of identified neonatal incertocortical neurons reveals a population of active cells that exhibit high frequencies of spontaneous action potentials and are capable of robustly activating neonatal cortical neurons. Because the fiber plexus is confined to layer 1, this pathway provides a spatially restricted excitatory GABAergic innervation of the distal apical dendrites of pyramidal neurons during the peak period of cortical synaptogenesis.

2-Amino-5-phosphonovalerate↗

PubMed1984

Ultrastructural peculiarities of the inner portion of the circular layer of colon. I. Research in the human.

The inner part of the circular layer of the human colon is made up of a special muscle layer, three to eight row thick. Wide areas of connective tissue are found inside this inner circular layer and between it and the outer part of the circular layer. The muscle cells of this innermost layer have several special features: numerous glycogen particles and caveolae, well-developed sarcoplasmic reticulum, and wide contact areas with each other. At the border zone with the tela submucosa, there is a fibrous lamella rich in collagen and elastic fibers, lined by an incomplete layer of circularly orientated fibroblast-like cells. A nerve plexus, rich in varicosities, is located between the two circular layers, and nerve fibers are sparsely located inside the fibrous lamella. Interstitial cells of Cajal were not recognized.

Adult↗

PubMed1987

Adrenergic receptors in inner and outer layers of human myometrium near term: characterization of beta-adrenergic receptor sites by [125I]-iodocyanopindolol binding.

The beta-adrenergic receptors present in inner and outer layers of human myometrium near term were characterized using the radioiodinated antagonist iodocyanopindolol (ICYP). In both layers ICYP binding is saturable, rapidly reversible, stereoselective, and appears to occur in a single class of sites with a KD of 30 pmol/l. Adrenergic agonists compete for ICYP binding sites with an order of potency consistent with beta 2-adrenergic potencies: isoproterenol greater than epinephrine much greater than norepinephrine. Studies in which CGP 20712 A, a beta 1-adrenergic antagonist, competes for ICYP binding sites in human myometrium reveal that at least 65% of the beta-receptors present are beta 2-subtype, whatever the layer considered. At the 35th week of pregnancy, the density of beta-adrenergic receptors in the inner layer (15.2 fmol/mg of protein) is about 50% higher than in the outer layer. At term, the densities of beta-adrenergic receptors are reduced and exhibit the same values for both layers (5 fmol/mg of protein). These results indicate that the beta-adrenergic receptors in the two layers diminish during pregnancy and reach, at term, an equal and low level of density.

Binding Sites↗

PubMed1992

Intravascular ultrasound imaging of coronary arteries. Is three layers the norm?

BACKGROUND: The purpose of this study was to evaluate the significance of the three-layered appearance of coronary arteries in adolescence and adults from intravascular ultrasound scans and to correlate these observations with histopathology. METHODS AND RESULTS: Sixteen intact hearts were excised at autopsy from patients with no clinical history of coronary artery disease. The patients' ages ranged from 13 to 55 years. A 30-MHz ultrasound imaging catheter was used to obtain images throughout the epicardial coronary vasculature. A total of 72 image cross sections was marked by epivascular sutures, and the corresponding histological sections were examined. Ultrasound images were classified into two groups: images exhibiting three-layered appearance and images without distinct layering. Histological analysis revealed a significantly greater degree of intimal thickening in segments with three layers (243 +/- 105 microns) than in nonlayered segments (112 +/- 55 microns). Discriminant analysis of these data predicted the threshold between the two groups to be 178 microns. Measurements of medial thickness were not different between these two groups (235 +/- 61 versus 210 +/- 76 microns). In the nonlayered group, the average patient age was 27.1 +/- 8.5 years, whereas in the three-layered groups, the average age was 42.8 +/- 9.8 years. CONCLUSIONS: The intracoronary ultrasound image appearance of young, morphologically normal coronary artery walls is homogeneous without layering. A three-layered appearance suggests the presence of at least 178 microns of intimal thickening and is seen more frequently with advancing age.

Adolescent↗

PubMed1979

High energy phosphate stores and lactate levels in different layers of the canine left ventricle during reactive hyperemia.

To study postischemic recovery of myocardial energy metabolism in relation to the reactive hyperemic response, the tissue levels of creatine phosphate (CP), adenosine triphosphate (ATP), and lactate were estimated in the outer, middle, and inner layers of the left ventricle before, during, and at several times after a 20-second bilateral coronary arterial occlusion in the open-chest dog. The reactive hyperemic response was characterized by monitoring blood flow in the cannulated coronary sinus in a separate group of dogs. Substantial changes in myocardial CP and lactate, but not ATP, were produced by the occlusion and reciprocal transmural gradients in CP and lactate occurred such that CP was lowest and lactate was highest in the inner layer. Recovery of high energy phosphate stores (CP + ATP) occurred long before blood flow returned to the preocclusion level, this being achieved in two of the three layers after completion of only 29% of the reactive hyperemic response when the blood flow debt repayment was 151%. Lactate returned to control levels during the response, but recovery times in the different layers were delayed compared to those for the high energy phosphate stores. A transmural lactate gradient was maintained as the regional levels declined, and recovery times were different for each of the three layers, being longest in the inner layer. The results suggest that during the reactive hyperemic response (1) kinetically different processes are involved in the repletion of energy stores and the removal of anaerobically produced metabolites, and (2) metabolic vasodilation of coronary vessels probably is more pronounced and sustained longer in the inner than in the outer ventricular layer.

Adenosine Triphosphate↗

PubMed1985

Variations in the functional electrical coupling between the subendocardial Purkinje and ventricular layers of the canine left ventricle.

Action potential propagation from the subendocardial Purkinje network into the ventricular muscle is an essential link in cardiac activation. Studies of papillary muscles have indicated that ventricular muscle activation by the Purkinje network occurs only at discrete, localized regions near the papillary muscle base. Over the rest of the endocardial surface, however, the spatial distribution of these subendocardial Purkinje to ventricular muscle connections has been less well defined. We therefore studied in vitro 12 canine left ventricular preparations (eight from the septum, four from the lateral wall), using a high-density (1-mm spacings), high-resolution extracellular mapping technique to determine the subendocardial Purkinje and ventricular muscle activation sequences. These studies show that the distribution of subendocardial Purkinje to ventricular muscle electrical coupling is spatially inhomogeneous, and that the junctional regions themselves have variable degrees of electrical coupling. We also attempted to determine whether ventricular muscle coupling to the Purkinje network might influence Purkinje network conduction velocity. We found that on the papillary muscle apex, a region without direct Purkinje to ventricular muscle propagation, Purkinje network conduction velocity was slowed, suggesting that the Purkinje network might be electrically loaded by the underlying ventricular muscle. Finally, we performed numerical simulations using a model consisting of two layers of excitable cells to evaluate the effects that different electrical coupling patterns and/or different coupling resistivities between the two layers might have on activation of each layer. These simulation studies suggest that a coupling pattern having discrete junctional sites between the two layers (similar to our findings for subendocardial Purkinje to ventricular muscle coupling) is beneficial, as this arrangement allows more rapid activation of both layers by minimizing electrical loading of the thin Purkinje layer by the thicker ventricular muscle layer.

Action Potentials↗

PubMed1986

Changes in plasma and buffy layer vitamin C concentrations following major surgery: what do they reflect?

Plasma and buffy layer vitamin C concentrations have been measured in 19 patients before and following major surgery, and falls of 36% and 43% respectively were recorded on the first post-operative day. However, the change in the buffy layer could not be accounted for by a change in the concentration of vitamin C in platelets, mononuclear cells or polymorphonuclear leucocytes, the cell types which form the buffy layer. The buffy layer change could be explained by a post-surgical increase in the number of polymorphonuclear leucocytes which, when the buffy layer vitamin C is expressed per 10(8) white cells, dilutes the platelet and mononuclear cell contribution to the buffy layer vitamin C concentration. The fall in buffy layer vitamin C does not, therefore, reflect any increased demand for white cell vitamin C in the immediate post-operative period, although the decrease in plasma vitamin C could imply some increased utilisation or redistribution of the non-cellular compartment.

Adult↗

PubMed1983

The ultrastructure and hardness of the transparent layer of human carious dentin.

The ultrastructure of the transparent layer of carious dentin was investigated in relation to hardness. This layer was the deeper part of the intermediately-softened inner carious dentin. Intratubular deposition of fine crystals was initially observed at the uppermost layer of normal dentin, increased in the subtransparent layer, and gradually shifted to deposition of rhomboid-shaped crystals in the transparent layer. Crystals were not seen in the tubules in the overlying discolored layer. Softening, due to demineralization of the intertubular and peritubular dentin, started at the bottom of the subtransparent layer and increased in the outward direction.

Crystallization↗

PubMedDate not supplied

Unstirred water layers in rabbit intestine: effects of guar gum.

Guar gum has been shown to affect the absorption of several different nutrients in clinical studies; however, the mechanisms for decreased absorption have not been defined. A possibility not studied with regard to guar gum, but previously demonstrated to be important in absorption, is the effect of change in the unstirred water layer. As the unstirred water layer increases in thickness, the rate of absorption decreases for certain nutrients. The effect of guar gum on the unstirred water layer in the lumen of rabbit jejunum was examined by previously described techniques. It was observed that: increases in guar gum concentration resulted in an increased thickness of the unstirred water layer; for any stir rate, the addition of guar gum increased the thickness of the unstirred water layer; and stir rate is inversely related to the thickness of the unstirred water layer. It was concluded from these results that guar gum increases the thickness of the unstirred water layer in rabbit jejunum. This mechanism may explain, in part, the reduction of the rate of absorption of certain nutrients seen following guar gum ingestion.

Animals↗

PubMedDate not supplied

The layering phenomenon and boundary formation in radiographs.

In a radiographic examination using a water-soluble contrast medium this may form a layer beneath a body fluid. Between the two liquids a zone consisting of a mixture of the two liquids then forms through diffusion. This diffusion layer produces some characteristic features in the radiographic image, an analysis of which was the purpose of the model experiments performed in this investigation. In this analysis of the layering phenomenon the radiographed objects were cylindrical tubes of methyl methacrylate partly filled with water. In some cases a rod was placed concentrically in the tube. Contrast medium was layered below the water. Radiographs were produced with the tube either vertical or inclined, and with either a horizontal or a vertical projection. In the image the layer of contrast medium was visualized as a light field, and the water layer as an overlying relatively dark field. The diffusion layer was visualized as a transitional zone--the diffusion field. Distinct boundaries and Mach lines observed in the bottom field were produced by the interface between the contrast medium and the solid wall where it was touched by the roentgen rays. These boundaries continued into the diffusion field where they gradually became less visible and eventually disappeared. The upper and lower boundaries of the diffusion field were diffuse and associated with dark and light Mach bands, respectively. The upper boundary appeared to be convex upwards. In the case of the inclined model and a vertical beam the diffusion field was elliptical, with a still more diffuse transition to the fields above and below than in the case of the vertical model and a horizontal beam.(ABSTRACT TRUNCATED AT 250 WORDS)

Body Fluids↗

PubMed1987

Distribution of activities of aspartate aminotransferase isoenzymes and malate dehydrogenase in guinea pig retinal layers.

Distributions of activity of the cytosolic (cAAT) and mitochondrial (mAAT) isoenzymes of aspartate aminotransferase and of malate dehydrogenase (MDH) were determined in guinea pig retinal layers. The distribution of total AAT activity (tAAT = cAAT + mAAT) and of mAAT activity correlated well (r = 0.88-0.91) with the distribution of MDH activity. mAAT activity was highest in the inner segments of the photoreceptors; there was a greater than twelve-fold difference between activity in that layer and in the inner retinal layers. cAAT activity was also highest in the inner segments, but the difference between the activity in the inner segments and the other layers was not nearly as great as with mAAT. cAAT activity was also relatively high in the outer nuclear layer, outer plexiform layer, and part of the inner plexiform layer. The high activity of cAAT, mAAT, and MDH in the inner segments indicates that all of these enzymes are involved in metabolic reactions related to energy production and/or to photoreceptive processes in the outer segments and, therefore, that the enzymes are probably involved in energy-related metabolism at synapses. However, other functions, including those related to neurotransmission, are not excluded.

Animals↗

PubMed2006

Single- versus two- layer intestinal anastomosis: a meta-analysis of randomized controlled trials.

BACKGROUND: To compare single- with two- layer intestinal anastomosis after intestinal resection: a meta-analysis of randomized controlled trials. METHODS: Randomized controlled trials comparing single- with two-layer intestinal anastomosis were identified using a systematic search of Medline, Embase and the Cochrane Library Databases covering articles published from 1966 to 2004. Outcome of primary interest was postoperative leak. A risk ratio for trial outcomes and weighted pooled estimates for data were calculated. A fixed-effect model weighted using Mantel-Haenszel methods and a random-effect model using DerSimonian-Laird methods were employed. RESULTS: Six trials were analyzed, comprising 670 participants (single-layer group, n = 299; two-layer group, n = 371). Data on leaks were available from all included studies. Combined risk ratio using DerSimonian-Laird methods was 0.91 (95% CI = 0.49 to 1.69), and indicated no significant difference. Inter-study heterogeneity was significant (chi2 = 10.5, d.f. = 5, p = 0.06). CONCLUSION: No evidence was found that two-layer intestinal anastomosis leads to fewer post-operative leaks than single layer. Considering duration of the anastomosis procedure and medical expenses, single-layer intestinal anastomosis appears to represent the optimal choice for most surgical situations.

Anastomosis, Surgical↗

PubMed2003

Cell clusters overlying focally disrupted mammary myoepithelial cell layers and adjacent cells within the same duct display different immunohistochemical and genetic features: implications for tumor progression and invasion.

INTRODUCTION: Our previous studies detected focal disruptions in myoepithelial cell layers of several ducts with carcinoma in situ. The cell cluster overlying each of the myoepithelial disruptions showed a marked reduction in or a total loss of immunoreactivity for the estrogen receptor (ER). This is in contrast to the adjacent cells within the same duct, which were strongly immunoreactive for the ER. The current study attempts to confirm and expand previous observations on a larger scale. METHODS: Paraffin sections from 220 patients with ER-positive intraductal breast tumors were double immunostained with the same protocol previously used. Cross-sections of ducts lined by > or = 40 epithelial cells were examined for myoepithelial cell layer disruptions and for ER expression. In five selected cases, ER-negative cells overlying the disrupted myoepithelial cell layer and adjacent ER-positive cells within the same duct were separately microdissected and assessed for loss of heterozygosity and microsatellite instability. RESULTS: Of the 220 cases with 5698 duct cross-sections examined, 94 showed disrupted myoepithelial cell layers with 405 focal disruptions. Of the 94 cases, 79 (84%) contained only ER-negative cell clusters, nine (9.6%) contained both ER-negative and ER-positive cell clusters, and six (6.4%) contained only ER-positive cell clusters overlying disrupted myoepithelial cell layers. Of the 405 disruptions, 350 (86.4%) were overlain by ER-negative cell clusters and 55 (13.6%) were overlain by ER-positive cell clusters (P < 0.01). Microdissected ER-negative and ER-positive cells within the same duct from all five selected cases displayed a different frequency or pattern of loss of heterozygosity and/or microsatellite instability at 10 of the 15 DNA markers. CONCLUSIONS: Cells overlying focally disrupted myoepithelial layers and their adjacent counterparts within the same duct displayed different immunohistochemical and molecular features. These features potentially represent an early sign of the formation of a biologically more aggressive cell clone and the myoepithelial cell layer breakdown possibly associated with tumor progression or invasion.

Breast↗

PubMed2005

Design and evaluation of 1- and 3-layer matrices of verapamil hydrochloride for sustaining its release.

The present study was performed to design oral controlled delivery systems for the water-soluble drug, verapamil hydrochloride, using natural and semisynthetic polymers as carriers in the forms of 1- and 3-layer matrix tablets. Verapamil hydrochloride 1-layer matrix tablets containing hydroxypropylmethylcellulose, tragacanth, and acacia either alone or mixed were prepared by direct compression technique. 3-layer matrix tablets were prepared by compressing the polymers as release retardant layers on both sides of the core containing the drug. The prepared tablets were subjected to in vitro drug release studies. Tragacanth when used as the carrier in the formulation of 1- and 3-layer matrices produced satisfactory release prolongation either alone or in combination with the other 2 polymers. On the other hand, acacia did not show enough prolonging efficiency in 1- and 3-layer matrix tablets. The results also showed that the location of the polymers in the 3-layer tablets has a pronounced effect on the drug release. Kinetic analysis of drug release from matrices exhibiting sustained release indicated that release was predominantly attributable to the contribution made by Fickian diffusion, while the erosion/relaxation mechanisms had a minor role in the release.

Chemistry, Pharmaceutical↗

PubMed2004

The receptor tyrosine kinase Off-track is required for layer-specific neuronal connectivity in Drosophila.

The nervous system in many species consists of multiple neuronal cell layers, each forming specific connections with neurons in other layers or other regions of the brain. How layer-specific connectivity is established during development remains largely unknown. In the Drosophila adult visual system, photoreceptor (R cell) axons innervate one of two optic ganglia layers; R1-R6 axons connect to the lamina layer, while R7 and R8 axons project through the lamina into the deeper medulla layer. Here, we show that the receptor tyrosine kinase Off-track (Otk) is specifically required for lamina-specific targeting of R1-R6 axons. Otk is highly expressed on R1-R6 growth cones. In the absence of otk, many R1-R6 axons connect abnormally to medulla instead of innervating lamina. We propose that Otk is a receptor or a component of a receptor complex that recognizes a target-derived signal for R1-R6 axons to innervate the lamina layer.

Aging↗