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The distribution and morphology of LGN K pathway axons within the layers and CO blobs of owl monkey V1.

The lateral geniculate nucleus (LGN) of primates contains three classes of relay cells, the magnocellular (M), parvocellular (P), and koniocellular (K) cells. At present, very little is known about either the structure or function of the K relay cells in New or Old World monkeys (simian primates). In monkeys, K cells are located between the main LGN layers and adjacent to the optic tract. For convenience, these intercalated cell layers are numbered K1-K4 starting closest to the optic tract with K1. The objective of this study was to examine the details of K axon morphology in the primary visual cortex (V1) of owl monkeys and to determine if different K layers give rise to distinct axon types. For this purpose, injections of WGA-HRP or PHA-L were made into specific K LGN layers and the distribution and morphology of the resulting labeled axons were analyzed. Injections of fluorescent tracers also were made within the superficial layers of V1 to further document connections via analysis of the patterns of retrogradely labeled cells in the LGN. Our main finding is that K axons in owl monkeys terminate as delicate focused arbors within single cytochrome oxidase (CO) blob columns in cortical layer III and within cortical layer I. Overall, the morphology of the K axons in these monkeys is quite similar to what we described previously for K geniculocortical axons in the distantly related bush baby (prosimian primate), suggesting that the basic features of this pathway are common to all primates. Our results also provide evidence that the axon arbors from different K layers are morphologically distinct; axons from LGN layer K1 project mainly to cortical layer I, while axons from LGN layer K3 chiefly terminate in cortical layer III. Taken together, these results imply that the basic features of axons within the K pathway are conserved across primates, and that the K axons from different K layers are likely to differ in function based upon their different morphologies.

Animals↗

Metal carboxylate-phosphonate hybrid layered compounds: synthesis and single crystal structures of novel divalent metal complexes with N-(phosphonomethyl)iminodiacetic acid.

Two novel divalent metal complexes with N-(phosphonomethyl)iminodiacetic acid, H(2)O(3)PCH(2)N(CH(2)CO(2)H)(2) (H(4)PMIDA), [Co(2)(PMIDA)(H(2)O)(5)] x H(2)O, 1, and [Zn(2)(PMIDA)(CH(3)CO(2)H)] x 2H(2)O, 2, have been synthesized and structurally characterized. The structure of complex 1 features two different kinds of Co(II) layers, namely, a cobalt phosphonate layer along the <100> plane and a cobalt carboxylate layer along the <300> plane. The Co(II) atoms in the phosphonate layer are octahedrally coordinated by 4 aqua ligands and 2 oxygen atoms from two phosphonic acid groups. Two Co(II) octahedra are bridged by a pair of phosphonic groups into a dimeric unit, and such dimers are interconnected into a layer through hydrogen bonding between aqua ligands. The Co(II) atoms in the carboxylate layer are octahedrally coordinated by a chelating PMIDA ligand, one aqua ligand, and one phosphonic oxygen atom from the neighboring PMIDA ligand. These Co(II) octahedra are interlinked by bridging carboxylic groups into a one-dimensional chain along the c-axis; such chains are held together by hydrogen bonds formed between carboxylic oxygen atoms and lattice water molecules, in such a way as to form a layer along the <300> direction. Two such layers are interconnected into a double layer via hydrogen bonding. These double layers are further interconnected with the Co(II) phosphonate layers through phosphonate tetrahedra along the a direction, resulting in the formation of a complicated three-dimensional network. The crystal structure of 2 contains a metal phosphonate and metal carboxylate hybrid layer along the <202> plane. One of the two zinc atoms in the asymmetric unit is tetrahedrally coordinated by four oxygen atoms from two phosphonic acid groups and two carboxylic groups; the other zinc atom is 5-coordinated by three oxygen atoms and a nitrogen atom from a chelating PMIDA ligand and one oxygen atom from the acetic acid. The above two types of zinc metal ions are interconnected by bridging carboxylic and phosphonic groups, resulting in the formation of a layered structure.

Journal Article↗

Analysis of the pressure-induced potential arising through composite membranes with selective surface layers.

When a pressure gradient is applied through a charged selective membrane, the transmembrane electrical potential difference, called the filtration potential, results from both the applied pressure and induced concentration difference across the membrane. In this work we investigate the electrokinetic properties relative to both active and support layers of a composite ceramic membrane close to the nanofiltration range. First, the volume charge density of the active layer is obtained by fitting a transport model to experimental rejection rates (which are controlled by the active layer only). Next, the value of the volume charge density is used to compute the theoretical filtration potential through the active layer. For sufficiently high permeate volume fluxes, the concentration difference across the active layer becomes constant, which allows assessing the membrane potential of the active layer. Experimental measurements of the overall filtration potential arising through the whole membrane are performed. The contribution of the support layer to this overall filtration potential is put in evidence. That implies that the membrane potential of the active layer cannot be deduced directly from the overall filtration potential measurements. Finally, the contribution of the support layer is singled out by subtracting the theoretical filtration potential of the active layer from the experimental filtration potential measured across the whole membrane (i.e., support + active layers). The amphoteric behavior of both layers is put in evidence, which is confirmed by electrophoretic measurements carried out with the powdered support layer and by recently reported tangential streaming potential measurements.

Journal Article↗

The ramification and connections of retinal fibres in layer 7 of the domestic chick optic tectum: a golgi impregnation, anterograde tracer and GABA-immunogold study.

Layer 7 is one of the retinorecipient layers of the avian optic tectum. However, little information is available about the neuronal organization of this layer and its implications for visual function. Golgi impregnation was used to investigate the retinal input to and the neuronal architecture of layer 7 of the chick optic tectum, which forms a narrow band between the two cell-dense layers 6 and 8. Anterograde tracers were also used to investigate the afferent and efferent connections of layer 7, in both the light and the electron microscope, together with GABA immunogold labelling. Three types of radial neuron were defined according to the origin and course of their axons. The perikarya of these neurons were situated in tectal layers 10-11. The principal dendrites of these radial neurons ascended to the tectal surface and gave rise to dendritic side-branches in layer 7. These dendritic side-branches received asymmetric synapses from the terminations of retinal fibre arborisations. Type 2 radial neurons, whose axons arose from the deep pole of the perikaryon or occasionally from a basal dendrite, were shown to project to the nucleus isthmi pars magnocellularis, which has previously been demonstrated to be GABAergic and to project to glomerulus-like complexes in tectal layers 4-5. In these layers, the dendritic branches of layer 13 neurons that project to the nucleus rotundus have previously been shown to receive retinal fibre input. Therefore, the retinal input to layer 7 may be able to modulate the transmission of information to the visual thalamus, by way of a feed-back loop to layers 4-5 of the tectum involving the nucleus isthmi pars magnocellularis.

Animals↗

Structural and functional MRI reveals multiple retinal layers.

MRI is a noninvasive diagnostic modality that reveals anatomy, physiology, and function in vivo without depth limitation or optical interference. MRI application to the retina, however, remains challenging. We improved spatial resolution to resolve layer-specific structure and functional responses in the retina and confirmed the laminar resolution in an established animal model of retinal degeneration. Structural MRI of normal rat retinas revealed three bands corresponding histologically to (i) the combined ganglion cell layer/inner nuclear layer plus the embedded retinal vessels, (ii) the avascular outer nuclear (photoreceptor) layer and its photoreceptor segments, and (iii) the choroidal vascular layer. Imaging with an intravascular contrast agent (gadolinium-diethylene-tri-amine-pentaacetic acid) enhanced the retinal and choroidal vascular layers bounding the retina, but not the avascular outer nuclear layer and the vitreous. Similarly, blood-oxygen-level-dependent (BOLD) functional MRI revealed layer-specific responses to hyperoxia and hypercapnia. Importantly, layer-specific BOLD responses in the two vascular layers were divergent, suggesting the two vasculatures are differentially regulated. To corroborate sensitivity and specificity, we applied layer-specific MRI to document photoreceptor degeneration in Royal College of Surgeons rats. Consistent with histology, layer-specific MRI detected degeneration of the outer nuclear layer. Surprisingly, MRI revealed increased thickness in the choroidal vascular layer and diminished BOLD responses to hyperoxia and hypercapnia in the Royal College of Surgeons rat retinas, suggesting perturbation of vascular reactivity secondary to photoreceptor loss. We conclude that MRI is a powerful investigative tool capable of resolving lamina-specific structures and functional responses in the retina as well as probing lamina-specific changes in retinal diseases.

Aging↗

The development of structure and function in the postnatal human cerebral cortex from birth to 72 months: changes in thickness of layers II and III co-relate to the onset of new age-specific behaviors.

HYPOTHESIS: The thickness of a cortical layer is a composite measure of neuronal, axonal, dendritic, synaptic, and glial numbers and sizes that may relate to thefunction of a cortical area. METHODS: 35 age-specific behaviors with defined cortical localization whose onset lies between birth and 72 months were selected. Each behavior's function localized to one or more of 12 cytoarchitectonic areas (Brodmann areas 4, with homuncular subdivisions for leg, trunk, face, and hand, plus 17, 18, 19, 20, 21, 24, 36, and 37). Data on cortical thickness for each layer of 41 cytoarchitectonic areas (including the 12 above) of the postnatal human cerebral cortex from birth of 72 months were analyzed for general patterns of change. For the 12 cortical areas functionally related to theage-specific behaviors, we searched for layer thickness changes that co-related to when the behaviors began. RESULTS: Without exception, all layers of the 41 cortical areas of the postnatal human cerebral cortex studied develop through a series of repeated thinning and thickening in a wave-like fashion. With regard to the co-relation of behavioral onset and changes in cortical layer thickness, from birth to 15 months, only layer II has agreater than expected frequency of being the layer with the greatest relative change in thickness (relative to its previous value). From 15 to 72 months, only layer IlI has a greater than expected frequency of being the layer with the greatest absolute change in thickness (81% involved a change in its direction of growth (thinning <--> thickening)). The co-occurrence of directional growth change and having the greatest layer thickness change were only statistically significant for layer III when an age-specific behavior began and was not seen for the 41 cortical areas overall (p = 0.014). CONCLUSIONS: Cortical laminar development exhibits aprocess that is mathematically consistent with a random walk with drift and with boundaries so that uncontrolled proliferation and pruning are prevented. The directional changes in layer growth could be controlled by feedback coupled with growth promoting and growth inhibiting factors. Layer II, with its function of establishing local corticocortical connections, appears to be most important in establishing age-specific behaviors of infants from birth to 15 months. Such a process tends to produce relatively simpler behaviors. LayerIII, with its function of establishing longer distance corticocortical connections, appears to be most important in establishing age-specific behaviors of children from 15 to 72 months. This process tends to produce richer, more cross-modal behaviors than those mediated primarily by local corticocortical interactions.

Aging↗

Morphology, electrophysiology and functional input connectivity of pyramidal neurons characterizes a genuine layer va in the primary somatosensory cortex.

Cortical layer V classically has been subdivided into sublayers Va and Vb on cytoarchitectonic grounds. In the analysis of cortical microcircuits, however, layer Va has largely been ignored. The purpose of this study was to investigate pyramidal neurons of layer Va in view of their potential role in integrating information from lemniscal and paralemniscal sources. For this we combined detailed electrophysiological and morphological characterization with mapping of intracortical functional connectivity by caged glutamate photolysis in layer Va of rat barrel cortex in vitro. Electrophysiological characterization revealed pyramidal cells of the regular spiking as well as the intrinsically burst firing type. However, all layer Va pyramidal neurons displayed uniform morphological properties and comparable functional input connectivity patterns. They received most of their excitatory and inhibitory inputs from intracolumnar sources, especially from layer Va itself, but also from layer IV. Those two layers were also the main origin for transcolumnar excitatory inputs. Layer Va pyramidal neurons thus may predominantly integrate information intralaminarly as well as from layer IV. The functional connectivity maps clearly distinguish layer Va from layer Vb pyramidal cells, and suggest that layer Va plays a unique role in intracortical processing of sensory information.

Animals↗

Dynamic properties of excitatory synaptic connections involving layer 4 pyramidal cells in adult rat and cat neocortex.

To investigate the properties of excitatory connections between layer 4 pyramidal cells and whether these differed between rat and cat, paired intracellular recordings were made with biocytin filling in slices of adult neocortex. These connections were also compared with those from layer 4 spiny cells to layer 3 pyramids and connections between layer 3 pyramids. Connectivity ratios for layer 4 pyramid-pyramid pairs (1:14 cat, 1:18 rat) appeared lower than for the other types of connections studied in parallel, but excitatory postsynaptic potential (EPSP) amplitudes and time course were not significantly different either between species or across types of connection. Layer 4 pyramids targeted postsynaptic basal dendrites in both species, whether the pyramidal target was in layer 4 or layer 3. Within layer 4, relationships between mean EPSP amplitude, numbers of putative contacts, and distance between connected pairs indicated a rapid decline in connectivity strength with distance, equivalent to 3.4 mV and 10 synapses per 100 microm separation, from a maximum of 4 mV and 10 synapses at 0 microm. However, a subset, of burst-firing layer 4 pyramids, appeared to make no connections with other layer 4 spiny cells. Second EPSPs were depressed by 36% in rat and 28% in cat relative to first EPSPs at interspike intervals <15 ms. Subsequent EPSPs in brief trains were further depressed. Depression was predominantly presynaptic in origin. Recovery from depression could not be described adequately by a simple exponential for individual connections; it included peaks and troughs with periodicities of 10-15 ms. Complex relationships between the first 2 interspike intervals and third EPSP amplitude were also apparent in all connections so studied. Large third EPSPs followed specific combinations of first and second interspike intervals so that increasing, or decreasing, one without changing the other resulted in a smaller third EPSP. Finally, the outputs of layer 4 spiny cells to layer 3 exhibited partial recovery from depression during longer high-frequency trains, a property not apparent in the other connections studied.

Animals↗

Importance of S-layer proteins in probiotic activity of Lactobacillus acidophilus M92.

AIMS: To investigate the functional role of surface layer proteins (S-layer) in probiotic strain Lactobacillus acidophilus M92, especially its influence on adhesiveness to mouse ileal epithelial cells. METHODS AND RESULTS: Sodium dodecyl sulphate polyacrylamide gel electrophoresis of cell surface proteins revealed the presence of potential surface layer (S-layer) proteins, ca at 45 kDa in L. acidophilus M92. Southern blot with pBK1 plasmid, containing slpA gene, gave a positive signal, suggesting that L. acidophilus M92 has a slpA gene coding for the S-layer proteins. S-layer proteins of this strain are present during all phases of growth. The S-layer proteins appeared when cells treated with 5 mol l(-1) LiCl were allowed to grow again. Removal of the S-layer proteins reduced adhesion of L. acidophilus M92 to mouse ileal epithelial cells. Furthermore, the viability of cells without S-layer were reduced in simulated gastric juice at low pH range (2, 2.5, 3) and simulated pancreatic juice with bile salts (1.5 and 3 g l(-1)). S-layer proteins of L. acidophilus M92 were resistant to pepsin and pancreatin, in contrast, the treatment with proteinase K led to a significant proteolysis of the S-layer proteins. CONCLUSIONS: These results demonstrated functional role of S-layer; it is responsible for adhesiveness of Lactobacillus acidophilus M92 to mouse ileal epithelial cells and has a protective role for this strain. SIGNIFICANCE AND IMPACT OF THE STUDY: S-layer proteins have an important role in the establishment of probiotic strain Lactobacillus acidophilus M92 in the gastrointestinal tract.

Animals↗

A four-layered 'lissencephalic' cortex induced by prenatal X-irradiation in the rat.

A four-layered cortex is produced in the dorsal region of the rat brain, after 200 cGy X-irradiation at embryonic day 16. The layer 1 is the molecular layer; layer 2 is the upper cellular layer which is in continuity with normal layers V and VI in the lateral cortex; layer 3 is a sparsely cellular layer; and layer 4 is the inner cellular layer which is composed of neurons normally committed to the upper cortical layers. This failure of neuroblast migration to the upper layers is associated with reduced numbers, and morphological abnormalities, of radial glial cells in the dorsal cortex. This further supports the idea that abnormal interactions between neuroblasts and radial glial cells result in abnormal migratory patterns. Since a similar four-layered organization is found in human lissencephaly type I, the present results bring new data that can serve our better understanding of human cortical malformations.

Abnormalities, Radiation-Induced↗

Surface display of foreign epitopes on the Lactobacillus brevis S-layer.

So far, the inability to establish viable Lactobacillus surface layer (S-layer) null mutants has hampered the biotechnological applications of Lactobacillus S-layers. In this study, we demonstrate the utilization of Lactobacillus brevis S-layer subunits (SlpA) for the surface display of foreign antigenic epitopes. With an inducible expression system, L. brevis strains producing chimeric S-layers were obtained after testing of four insertion sites in the slpA gene for poliovirus epitope VP1, that comprises 10 amino acids. The epitope insertion site allowing the best surface expression was used for the construction of an integration vector carrying the gene region encoding the c-Myc epitopes from the human c-myc proto-oncogene, which is composed of 11 amino acids. A gene replacement system was optimized for L. brevis and used for the replacement of the wild-type slpA gene with the slpA-c-myc construct. A uniform S-layer, displaying on its surface the desired antigen in all of the S-layer protein subunits, was obtained. The success of the gene replacement and expression of the uniform SlpA-c-Myc recombinant S-layer was confirmed by PCR, Southern blotting MALDI-TOF mass spectrometry, whole-cell enzyme-linked immunosorbent assay, and immunofluorescence microscopy. Furthermore, the integrity of the recombinant S-layer was studied by electron microscopy, which indicated that the S-layer lattice structure was not affected by the presence of c-Myc epitopes. To our knowledge, this is the first successful expression of foreign epitopes in every S-layer subunit of a Lactobacillus S-layer while still maintaining the S-layer lattice structure.

Bacterial Proteins↗

Isolation and in vitro assembly of the components of the outer S layer of Lampropedia hyalina.

The outermost component of the S layer of Lampropedia hyalina, the punctate layer, is assembled onto an inner perforate layer. The punctate layer is composed of long, tapered cylindrical units centered on p6 symmetry axes and connected by six fine linking arms, joining at the axis of threefold symmetry to create a hexagonal layer with a lattice constant of 25.6 +/- 0.5 nm (J. A. Chapman, R. G. E. Murray, and M. R. J. Salton, Proc. R. Soc. London Ser. B 158:498-513, 1963; R. G. E. Murray, Can. J. Microbiol. 9:593-600, 1963). Extraction of cell envelopes with 100 mM Tris buffer (pH 8) containing 2% deoxycholate resulted in the release of several proteins, but left the S layers intact. The punctate layer was then extracted with 3 M guanidine hydrochloride or 6 M urea, leaving the perforate layer intact. This treatment led to the release of three polypeptides with molecular weights of 60,000, 66,000, and 240,000 (60K, 66K, and 240K polypeptides). These three polypeptides reassembled on the perforate layer as a template to form the S-layer complex or self-assembled to form the punctate layer alone after dialysis of the extract against 50 mM HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) buffer (pH 7.5) containing 10 mM CaCl2. The self-assemblies were composed of a 240K polypeptide and a 60K polypeptide. The 240K and 60K polypeptides were separated by column chromatography and examined by electron microscopy. The 240K polypeptide appeared in negative stain as a long, flexible structure and assembled into loose arrays with sixfold symmetry with obvious Y-shaped linking elements, while fractions containing both the 60K and 240K polypeptides showed assemblies closely resembling the punctuate layer. Immunoelectron microscopy was used to confirm the presence of both the 60K and 240K polypeptides as components of the punctuate layer.

Cell Fractionation↗

Influence of the secondary cell wall polymer on the reassembly, recrystallization, and stability properties of the S-layer protein from Bacillus stearothermophilus PV72/p2.

The high-molecular-weight secondary cell wall polymer (SCWP) from Bacillus stearothermophilus PV72/p2 is mainly composed of N-acetylglucosamine (GlcNAc) and N-acetylmannosamine (ManNAc) and is involved in anchoring the S-layer protein via its N-terminal region to the rigid cell wall layer. In addition to this binding function, the SCWP was found to inhibit the formation of self-assembly products during dialysis of the guanidine hydrochloride (GHCl)-extracted S-layer protein. The degree of assembly (DA; percent assembled from total S-layer protein) that could be achieved strongly depended on the amount of SCWP added to the GHCl-extracted S-layer protein and decreased from 90 to 10% when the concentration of the SCWP was increased from 10 to 120 microg/mg of S-layer protein. The SCWP kept the S-layer protein in the water-soluble state and favored its recrystallization on solid supports such as poly-L-lysine-coated electron microscopy grids. Derived from the orientation of the base vectors of the oblique S-layer lattice, the subunits had bound with their charge-neutral outer face, leaving the N-terminal region with the polymer binding domain exposed to the ambient environment. From cell wall fragments about half of the S-layer protein could be extracted with 1 M GlcNAc, indicating that the linkage type between the S-layer protein and the SCWP could be related to that of the lectin-polysaccharide type. Interestingly, GlcNAc had an effect on the in vitro self-assembly and recrystallization properties of the S-layer protein that was similar to that of the isolated SCWP. The SCWP generally enhanced the stability of the S-layer protein against endoproteinase Glu-C attack and specifically protected a potential cleavage site in position 138 of the mature S-layer protein.

Bacterial Proteins↗

Three-dimensional mechanical properties of porcine coronary arteries: a validated two-layer model.

The normal coronary artery consists of two mechanically distinct layers: intima-media and adventitia. The objective of this study is to establish a two-layer three-dimensional (3-D) stress-strain relation of porcine coronary arteries. Experimental measurements were made by a series of biaxial tests (inflation and axial extension) of intact coronary arteries and, subsequently, their corresponding intima-media or adventitia layer. The Fung-type exponential strain energy function was used to describe the 3-D strain-stress relation for each layer and the intact wall. A genetic algorithm was used to determine the material constants in the Fung-type constitutive equation by curve fitting the experimental data. Because one layer must be sacrificed before the other layer can be tested, the material property of the missing layer was computed from the material constants of the intact vessel and the tested layer. A total of 20 porcine hearts were used: one group of 10 hearts for the left anterior descending artery and another group of 10 hearts for the right coronary artery. Each group was further divided into two subgroups of five specimens tested for the intact wall and the intima-media layer and for the intact wall and the adventitia layer. Our results show statistically significant differences in the material properties of the two layers. The mathematical model was validated by experimental stress-strain data for individual layers. The validated 3-D constitutive model will serve as a foundation for formulation of layer-specific boundary value problems in coronary physiology and cardiology.

Algorithms↗

W-and Y-cells in the C layers of the cat's lateral geniculate nucleus: normal properties and effects of monocular deprivation.

1. Previous studies have shown that rearing with monocular visual deprivation (MD) produces a loss of Y-cells and a reduction in spatial resolution among X-cells in layers A and A1 of the cat's dorsal lateral geniculate nucleus (dLGN). However, there have been no studies of the effects of visual deprivation on the function of the retinogeniculate W-cell pathway, which terminates in the C layers of the dLGN. It also is not known if Y-cells in the C layers are affected by MD in the same way as Y-cells in the A layers. These questions were addressed by the present experiment. 2. Single-cell recordings were made from the C layers of 5 normal adult cats (112 cells) and from the nondeprived (94 cells) and deprived (95 cells) C layers in 10 cats monocularly deprived by lid suture for 3-7 yr. The cells were classified as X, Y, or W on the basis of their receptive-field properties and responses to electrical stimulation of the optic chiasm. In addition, quantitative measures were made of responses to sine-wave gratings of different spatial frequencies. 3. Receptive-field organization, receptive-field center size, spatial and temporal linearity to counterphased sine-wave gratings, and latency to optic chiasm stimulation were similar for C-layer cells in normal cats and in the deprived and nondeprived layers of MD cats. On the basis of these properties, 23% of normal layer-C cells were classified as Y-cells and 72% were classified as W-cells. The Y-cells tended to be located in the magnocellular division of layer C and most (though not all) W-cells were in the parvocellular division. Normal layers C1 and C2 contained almost exclusively W cells. The incidence of Y and W cells was similar to normal in the nondeprived and deprived C-layers of MD cats. 4. In normal cats, W cells typically had the lowest amplitude first-harmonic (F1) response rates to drifting sine-wave gratings. However, many W cells gave quite brisk responses and, overall, there was no significant difference between F1 response amplitudes of Y and W cells. Response amplitudes of Y- and W-cells in the deprived and nondeprived C-layers of MD cats were not significantly different from normal. 5. Normal Y- and W-cells tended to have low optimal spatial frequencies (0.2 c/deg or lower) and spatial resolutions (generally 0.4-1.6 c/deg) to drifting sine-wave gratings.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Sustained-release phenylpropanolamine hydrochloride bilayer caplets containing the hydroxypropylmethylcellulose 2208 matrix. II. Effects of filling order in bilayer compression and manufacturing method of the prolonged-release layer on compactibility of bilayer caplets.

The purpose of this study was to establish the manufacturing method of the formulated bilayer caplets containing the hydroxypropylmethylcellulose 2208 (HPMC2208) matrix without lamination. In manufacturing the bilayer caplets containing the HPMC2208 (Metolose 90SH4000) matrix, some bilayer caplets were cracked. We found that cracking of bilayer caplets is not the separation of two layers, but lamination of the prolonged-release layer. It was assumed that Metolose 90SH4000 causes lamination of the prolonged-release layer. Two factors, roller compaction pressure on dry granulation of the prolonged-release layer and filling order of the prolonged-release layer in bilayer compression, were related to lamination of bilayer caplets. The compactibility of the prolonged-release layer decrease with an increase in roller compaction pressure on dry granulation. The compactibility of the prolonged-release layer manufactured by direct compression is superior to that manufactured by dry granulation. The compactibility of the prolonged-release layer in the shape of the second layer, convexo-concave, is superior to that in the shape of the first layer, convexo-convex. This is due to the fact that the density distribution inside the compact in the shape of convexo-concave was more uniform than that in the shape of convexo-convex. The manufacturing method of the formulated bilayer caplets having the prolonged-release layer whose Metolose 90SH4000 content is 60% without lamination is as follows: the prolonged-release layer manufactured by direct compression is fed as the second layer in bilayer compression.

Chemistry, Pharmaceutical↗

Effects of graded coronary constriction on regional oxygen and carbon dioxide tensions in outer and inner layers of the canine myocardium.

This study was undertaken to investigate the effects of graded coronary constriction on regional gas tensions of the myocardium. In 12 open chest dogs, tissue carbon dioxide (PtCO2) and oxygen (PtO2) tensions were measured simultaneously in outer and inner layers of the myocardium using a mass spectrometer. In normal condition, higher PtO2 and lower PtCO2 were observed in outer layer than in inner layer. With application of coronary constriction, increase in PtCO2 and decrease in PtO2 were observed in both layers of the myocardium, but the response to the ischemic stimuli by applying coronary constriction in inner layer was different from that in outer layer. Severe coronary constriction, more than 90% in its diameter, was necessary to produce significant changes in both gas tensions in both layers of the myocardium. Decrease in PtO2 was found in the condition of less severe coronary constriction and to be greater in inner layer than in outer layer of the myocardium. In terms of the changes in PtCO2, inner layer was also more susceptible to the ischemic stimuli than outer layer. The greater and earlier elevation of PtCO2 in inner layer than in outer layer is regarded as one of the possible mechanisms of the reduction of myocardial contraction in the early stage of myocardial ischemia.

Animals↗

Atrophy of relay neurons in magno- and parvocellular layers in the lateral geniculate nucleus in experimental glaucoma.

PURPOSE: To determine whether in glaucoma there is atrophy of relay neurons in magnocellular and/or parvocellular lateral geniculate nucleus (LGN) layers projecting to the visual cortex and to compare the degree of neuronal atrophy in magnocellular layers with that in parvocellular layers. METHODS: Seven cynomolgus monkeys with unilateral experimentally induced glaucoma and five control monkeys were studied. The left LGN neurons in magnocellular layer 1 and parvocellular layers 4 and 6, connected to the right glaucomatous eye were examined. Immunocytochemistry with antibody to parvalbumin was used to specifically label relay neurons connecting to the visual cortex. Neuronal cell body cross-sectional area was estimated using unbiased point-counting methodology. Experimental and control groups were compared using t-tests. Analysis of covariance (ANCOVA) tests were used to compare the percentage of decrease in mean neuronal area between layers 1, 4, and 6, as a function of percentage of optic nerve fiber loss or mean IOP. There was significant correlation between percentage of optic nerve fiber loss and mean IOP. RESULTS: The mean cross-sectional area of relay neurons in magnocellular layer 1 and parvocellular layers 4 and 6 were significantly decreased in glaucoma compared with controls by 28%, 37%, and 45%, respectively. Neuronal area decreased in a linear fashion, with increasing optic nerve fiber loss or increasing mean IOP for layers 1, 4, and 6. The percentage of neuronal shrinkage in each of parvocellular layers 4 and 6, as a function of optic nerve fiber loss (P = 0.05; P = 0.001, respectively) or mean IOP (P = 0.046; P = 0.0008, respectively), was greater than that seen in magnocellular layer 1. CONCLUSIONS: Relay neurons in the LGN, which project to the visual cortex, undergo significant shrinkage in glaucoma, and neurons in parvocellular layers undergo significantly more shrinkage than neurons in magnocellular layers.

Animals↗