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Epileptiform activity can be initiated in various neocortical layers: an optical imaging study.

The initiation site for triggering epileptiform activity was investigated via optical imaging using voltage-sensitive dyes in the neocortical slice perfused with artificial cerebral spinal fluid containing nominally zero magnesium. The neocortical slices (400-microm thick) were harvested from Sprague-Dawley rats (P21-28). Optical imaging was made by using a high speed photodiode array. Spontaneous epileptiform activity emerged 20-40 min after the preparation was perfused with zero-magnesium solution. There was a good correspondence between electrical and optical signals (n = 46), although the details of the two recordings were somewhat different. The initiation sites were measured optically in 11 preparations. Among them, four were found to be located in superficial layers, two were found in middle layers, and five were found in deep layers. Repeated recordings revealed that these initiation sites were relatively stable; shifting of the initiation site was not observed. Therefore spontaneous epileptiform activity could be initiated in various cortical layers, from layer I to layer VI. The activation started from a small area <0.04 mm(3) and spread smoothly from the initiation site to adjacent cortical areas, suggesting that the initiation site is very confined to one of the cortical layers. The initiation sites were distributed randomly in various cortical areas, and no higher probability was found in a special cortical region. Electrical stimulation delivered via a glass microelectrode filled with 2 M NaCl (2-5 MOhms) could reliably trigger epileptiform activity that had the same characteristics as the spontaneous activity. The cortical neurons activated directly by the stimulation were around the electrode's tip and estimated to be within a 50-microm area, suggesting that only a few neurons were needed to form an initiation site. Because the timing for stimulation was arbitrary and the evoked events were initiated independent of discharges of neurons in any other layers, it is likely that the initiation site for epileptiform activity in various cortical layers is independent of the control of layer V pyramidal neurons. Together these finding suggest that the epileptiform focus is confined and can be formed in several (probably all) neocortical layers and in many cortical areas. The initiating neurons may be of different types because neuronal types in various cortical layers are different.

Animals↗

Influence of a fat on muscle oxygenation measurement using near-IR spectroscopy: quantitative analysis based on two-layered phantom experiments and Monte Carlo simulation.

The influence of a subcutaneous fat layer on measurement of muscle oxygenation using near-IR spectroscopy was quantitatively investigated by two-layered phantom experiments and Monte Carlo simulations, with the aim of developing an algorithm that can correct this influence. The phantom consisted of a fat-like layer, which was a mixture of agar and titanium dioxide powder, and a muscle-like layer, which was suspension of washed bovine blood in Intralipid solution. An LED with 760 and 840 nm elements was used as an optical source, and the backscattered light was detected by photodiodes at source-detector distances of 20, 30 and 40 mm. The relationships between changes in optical density and blood concentrations were obtained at fat-like layer thicknesses of 0.5,10 and 15 mm under fully oxygenated and fully deoxygenated states. It was experimentally found that the change in optical density is significantly decreased and the linearity of measurement characteristics is clearly distorted by the presence of a fat layer. In the simulations, normalized light reflectance and mean optical pathlength in a muscle layer were calculated. The simulation results of the light reflectance agreed well with the experimental results. When the absorption in a muscle layer was relatively high, the mean optical pathlength in the muscle layer, or the measurement sensitivity, was not so dependent on the absorption. Therefore, the modified Beer-Lambert law can still be applied to estimate changes in muscle absorption from changes in optical density, even when a fat layer is involved. The results of simulation also suggested that the influence of a fat layer can be eliminated by correcting the measurement sensitivity using the fat layer thickness.

Adipose Tissue↗

Drug permeation through the three layers of the human nail plate.

The in-vitro permeation characteristics of a water soluble model drug, 5-fluorouracil, and a poorly water soluble model drug, flurbiprofen, were investigated through three layers of the human nail plate (namely, the dorsal, intermediate and ventral nail plates), using a modified side-by-side diffusion cell. The dorsal-filed nail plate, the ventral-filed nail plate and the dorsal-and-ventral-filed nail plate were prepared to known thicknesses and then used with the full-thickness nail plate to investigate the permeation characteristics of each single layer. Most of the lipids in the human nail plate were found in the dorsal and ventral layers. The rank orders of the permeation fluxes for 5-fluorouracil and flurbiprofen were both: dorsal-and-ventral-filed nail plate > dorsal-filed nail plate > ventral-filed nail plate > full-thickness nail plate. With respect to 5-fluorouracil permeation through each single layer, the permeability coefficient of the intermediate layer was higher than those of other single layers. However in the case of flurbiprofen, the permeability coefficient of the ventral layer was higher than other single layers. The diffusion coefficients of 5-fluorouracil and flurbiprofen in the dorsal layer were the lowest of any single layer. The drug concentration in each layer was estimated using each respective permeation parameter. The drug concentration in the nail plate was observed to be dependent on the solubility and the flux of the drug. From these findings, we suggest that the human nail plate behaves like a hydrophilic gel membrane rather than a lipophilic partition membrane and that the upper layer functions as the main nail barrier to drug permeation through its low diffusivity against the drugs.

Diffusion↗

Design of controlled release system with multi-layers of powder.

Pellets containing active ingredients were coated with water-insoluble powders, i.e. hydrogenated caster oil (Lubliwax (WAX)) and magnesium stearate (Mg-St). The influences of the structural difference of the sustained release layer and curing conditions on the drug release rate were investigated. Sodium valproate (VP-Na) was used as a highly water-soluble model drug. Drug release profiles were influenced by the combination of the WAX layer and the Mg-St layer. Even if the formula of sustained release layers were the same, drug release rate could be affected by the structural difference of the controlled release layer. The Mg-St layer was more effective in prolonging drug release than the WAX layer. Compared with single and double layer types, the triple layer (sandwich) type was most effective in obtaining a long sustained drug release. Heat-treatment retarded drug release mainly by increasing the density of the sustained release layer of the WAX. The Mg-St was effective in protecting the agglomeration between particles during heat-treatment. Optimal heat-treatment conditions were found to exist. Scanning electron microscopy (SEM) analysis indicated that heat-treatment caused the WAX to melt, formed a film-like structure and made the release layer dense. Furthermore, heat-treatment changed the release pattern of VP-Na from sustained release pellets with a multi-layer of powder, leading to zero-order release.

Delayed-Action Preparations↗

Supporting layers of the glenohumeral joint. An anatomic study.

Based on anatomic and surgical dissections, the anatomy of the shoulder region is described in terms of four layers that overlie and support the glenohumeral joint. Each layer envelops the glenohumeral joint on its anterior, lateral, and posterior aspects, and between each layer there is a plane for safe and easy dissection. Layer 1 is composed of the deltoid and pectoralis major muscle bellies with their overlying fascia and enveloping epimysium. Anteriorly, Layer 2 consists of the clavipectoral fascia, the conjoined tendon of the short head of the biceps and coracobrachialis, and the coracoacromial ligament. Posteriorly, Layer 2 is the dense posterior scapular fascia that overlies the infraspinatus and teres minor muscle bellies. It is continuous with the clavipectoral fascia around the lateral aspect of the proximal humerus. Deep to Layer 2, the subdeltoid bursa yields a dissection plane that encompasses the anterior, lateral, superior, and variably the posterior aspects of the glenohumeral joint. Layer 3 consists of the deep layer of the subdeltoid bursa and the underlying musculotendinous units of the rotator cuff, including subscapularis, supraspinatus, infraspinatus, and teres minor. Layer 4 is the capsule of the glenohumeral joint. This includes the glenohumeral ligaments and coracohumeral ligament. These four layers were present and consistent in each shoulder dissected. Significant variations were present only within the deepest layer (shoulder joint capsule). This system can serve as a learning tool and will provide a more organized approach to facilitate surgical dissection in the region.

Cadaver↗

Growth of the two layers of the chick sclera is modulated reciprocally by visual conditions.

PURPOSE: Although visual deprivation causes increased ocular elongation and myopia in both birds and mammals, changes in sclera appear to be in opposite directions. Because avian sclera has a cartilaginous layer as well as the fibrous layer found in mammals, we examined whether the scleral responses to various visual manipulations differ between the two layers. METHODS: To produce increases in ocular elongation and myopia, monocular diffusers or negative lenses were fitted to eyes. Conversely, to produce decreases in ocular elongation, diffusers were removed (restoring normal vision) or monocular positive lenses were fitted. Scleral layers were then dissected apart, and incorporation of labeled precursors into glycosaminoglycans (GAGs), DNA, and protein was assessed. Tissue coculture experiments were used to assess humoral interactions between scleral layers and with the choroid. RESULTS: In the cartilaginous layers, the incorporation of label into proteoglycans and DNA was significantly higher in eyes elongating faster than normal because of wearing diffusers or negative lenses and significantly lower than normal in eyes elongating slower than normal because of removal of the diffuser or wearing positive lenses. In the fibrous layers, the reverse was the case. Coculturing cartilaginous sclera from normal eyes with fibrous sclera from myopic or recovering eyes produced the same increase or decrease in sulfate incorporation into GAGs in the cartilaginous layer as though the tissue measured was from the animal providing the conditioning tissue. Coculturing with choroid, especially from recovering eyes, also inhibited cartilaginous sclera. CONCLUSIONS: The fibrous layer of the avian sclera shows changes in sulfate incorporation into GAGs during deprivation and recovery from deprivation in the same direction as does the mammalian sclera, whereas the cartilaginous layer changes in the opposite direction. The responses of the cartilaginous layer may be controlled by the fibrous layer, although they are influenced by the choroid as well.

Animals↗

Backward cortical projections to primary somatosensory cortex in rats extend long horizontal axons in layer I.

We have studied the origin and extent of axons within layer I of the primary somatosensory cortex (SI) of rats by using retrograde and anterograde tracers with an emphasis on reciprocal connections to layer I of SI from ipsilateral cortical areas that are the target of SI projections. Small crystals of 1,1',dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate (DiI) labeled horizontal axons projecting in all directions within layer I, which extended for up to 4 mm with numerous terminal branches. Applications of horseradish peroxidase, Diamidino yellow, or fast blue to the pial surface of SI labeled a characteristic pattern of neurons below the application site that excluded neurons in layer IV of the barrel fields, unless the dye penetrated deeper than layer II. This provided a control for the effective depth of the layer I dye applications. Retrograde transport from layer I of SI was traced to the primary motor area, the lateral parietal areas, including the secondary somatosensory (SII) and agranular insular cortex ipsilaterally, as well as the homotopic areas of SI contralaterally. Injections of the anterograde tracer dextran amine at the same site as the SI surface application labeled dense fiber terminations in middle layers of these same secondary areas in the primary motor cortex (MI) or SII in the midst of cells labeled by retrograde transport from layer I of SI. Injections of dextran amine into these secondary cortical areas labeled fibers that coursed through deep layers to SI, where they ascended to layer I. These reciprocal corticocortical inputs to SI were concentrated in layer I, where they branched and extended horizontally across several SI barrels.

Animals↗

The effect of smear layer on microbial coronal leakage of gutta-percha root fillings.

The aim of this in vitro study was to determine the effect of removal of the smear layer on canal obturation as measured by penetration of bacteria from a coronal direction. One hundred and twenty extracted human teeth with straight, single root canals were decoronated. The canals were prepared using the modified double-flared technique with balanced force under copious irrigation. The apical matrix was prepared to size 40 and apical patency subsequently confirmed with a size 15 file. The teeth were divided randomly into experimental groups (80 teeth) and control groups (40 teeth). The root canals of 40 experimental and 20 control teeth were rinsed with 40% citric acid and 2% NaOCl to remove the smear layer before obturation. In experimental groups, 20 teeth with smear layer intact and 20 teeth with smear layer removed were obturated with lateral condensation of cold gutta-percha and Apexit sealer. A further 20 teeth with smear layer intact and 20 teeth with smear layer removed were obturated with the Trifecta technique with the same sealer. In control groups, 10 teeth with smear layer intact and 10 teeth with smear layer removed were obturated with lateral condensation of cold gutta-percha and Apexit sealer. These teeth were completely sealed both coronally and apically to serve as negative controls. The remaining 20 teeth with either smear layer intact or smear layer removed were not obturated and served as the positive controls. The root surface of each tooth was sealed with nail varnish. The cut end of a polypropylene tube was sealed around the coronal part of each root canal so that bacteria placed therein could move only through the obturated canal space. Each root was placed in a glass bottle containing sterile Todd-Hewitt Broth (THB) and aliquots of 0.5 ml of THB were injected into the polypropylene tube. The model system was centrifuged at 168 g. An innoculum of Streptococcus sanguis in THB was placed in each coronal chamber at 5-day intervals and daily observations were made for bacterial growth in the apical reservoir for 90 days. All positive control teeth showed bacterial penetration within 24 h, while the negative control teeth remained uncontaminated throughout the test period. Leakage through the experimental teeth was variable ranging from 7 to 86 days. There was no statistical significant difference (P > 0.05) in leakage between the obturated canal when the smear layer was either removed or intact.

Calcium Hydroxide↗

Projection of the nucleus pretectalis to a retinorecipient tectal layer in the pigeon (Columba livia).

The avian optic tectum is composed of at least 15 separate laminae that are distinguishable on the basis of their morphological features and patterns of afferent and efferent connectivity. Layer 5b, a major retinorecipient layer, exhibits dense, dust-like, neuropeptide Y-positive (NPY+) immunoreactive labeling, whereas sparse, larger caliber NPY+ fibers are found in laminae 4 and 7. Anterograde and retrograde labeling techniques, immunohistochemistry, and retinal lesion studies were used to determine the source of this tectal NPY+ labeling. NPY+ was not detectable in cells of the optic tectum or in retinal ganglion cells, and retinal ablation did not diminish the abundance of tectal NPY+ fibers. Neurons of two nuclei previously shown to be sources of tectal input, the nucleus pretectalis (PT) and the intergeniculate leaflet (IGL; Brecha, 1978), were found to be NPY+. Unilateral injection of retrograde tracers into the tectum resulted in bilateral labeling of neurons within PT, and injections of anterograde tracer into PT confirmed that this nucleus projected bilaterally to layer 5b of the optic tectum. Unilateral lesions of PT nearly eliminated NPY+ fibers in the ipsilateral layer 5b and significantly reduced them in the contralateral layer 5b. Bilateral lesions of PT eliminated NPY+ fibers bilaterally in layer 5b. However, these PT lesions had little effect on the NPY+ fibers in layers 4 and 7. Combined retrograde and immunohistochemical studies showed that NPY+ neurons of the IGL project to the optic tectum, and anterograde studies demonstrated that IGL projects to layers 4 and 7. The NPY+ projection to laminae 5b from PT is one of many inputs, which include cholinergic afferents from the nucleus isthmi parvicellularis, terminals from retinal ganglion cells, and dendrites of layer 13 neurons (Karten et al., 1993). The NPY+ input to layer 5b may modulate visual information flow from retinal input to various tectal neurons, including those in layer 13.

Animals↗

Retinal ganglion cell projections to individual layers of the lateral geniculate body in Galago crassicaudatus.

The genus Galago provides an unique opportunity to study the relation between layers of the lateral geniculate body and classes of retinal ganglion cells. In the present experiments HRP was restricted to individual layers of the lateral geniculate body with the following results: After injections of the magnocellular layers, layers 1 and 2, labeled retinal ganglion cells ranged in size from 8 to 20 micrometers. After injections of the parvocellular layers, layers 3 and 6, labeled retinal ganglion cells ranged in size from 6 to 12 micrometers. After injections involving layers 4 and 5, which layers contain only very small, pale cells, labeled retinal ganglion cells ranged in size from 5 to 14 micrometers. Thus, the very largest ganglion cells were labeled only after injections of magnocellular layers 1 and 2, while small and medium retinal ganglion cells were labeled after HRP injections in every layers of the lateral geniculate body. Because the magnocellular layers actually contain a mixture of large, medium, and small-sized cells, we suggest that retinal ganglion cells of different size-classes project to geniculate relay cells of the corresponding size-class.

Animals↗

Geniculocortical projection to layer I of area 17 in kittens: orthograde and retrograde HRP studies.

Using the orthograde and retrograde HRP methods, the distribution of geniculocortical (G-C) afferent terminals in area 17 was examined in kittens and an adult cat. Following HRP injection in the entire dorsal lateral geniculate nucleus (LGNd), afferent terminals were labeled in layer I and deeper layers. The terminals in layer I in young kittens (less than 2 weeks old) were densely distributed throughout layer I, whereas in old kittens (over 1 month old) they were sparsely distributed in the outer half of layer I. When the HRP injection site was confined to the A laminae (lamina A and lamina A1), a few HRP-labeled terminals in layer I were seen in young kittens but not in older kittens or an adult cat. Following HRP injection in layer I of area 17 in young kittens, relatively large neurons were labeled retrogradely. The labeled neurons were located chiefly in the C complex and partly in the A laminae. The present experiments revealed that the G-C afferents to layer I of area 17 in young kittens originate from relatively large neurons which are located chiefly in the C complex and partly in the A laminae and that these afferents give dense terminals covering the entire layer I. These findings contrast with those obtained in adult cats, in which the G-C afferents to layer I originate exclusively from small neurons in the C complex and give sparse terminals to the outer half of layer I.

Animals↗

Dendritic bundling in layer I of granular retrosplenial cortex: intracellular labeling and selectivity of innervation.

The extrinsic projections to and from the retrosplenial cortex have been studied in detail, but the intrinsic circuitry within this region has been characterized less completely. To further define the internal connections, small injections of the retrograde, fluorescent tracer Fluorogold were made into the retrosplenial cortex of the rat. These injections label neurons in layers II-V of the contralateral homotopic cortex. In layers III-V, the labeled neurons are present over an area much larger than the injection site, but in layer II neurons are labeled in a very precise homotopic pattern. Following these injections, only the neurons in layer II display heavily labeled apical dendrites, and these labeled dendrites form tight bundles in layer Ic and Ib of the cortex and spread out in layer Ia. An examination of Golgi-stained material demonstrates that most of the neurons in layer II are small pyramidal cells with 2-3 small basal dendrites and a single, large apical dendrite that arborizes extensively in layer Ia. To verify the structure of the layer II neurons, they were intracellularly filled with Lucifer yellow. Examination of these labeled cells confirms the observations from the Golgi-stained material and demonstrates that many apical dendrites of the layer II cells angle acutely, apparently to join a bundle and/or avoid an interbundle space. Tract tracing experiments demonstrate that the anteroventral nucleus of the thalamus appears to project selectively to the region containing the dendritic bundles, whereas intracortical projections appear to terminate in layers Ib and Ic in the 30-200 microns spaces between the bundles. Furthermore, the areas containing the bundles display dense AChE staining, but the interbundle spaces are almost free of AChE staining. These findings demonstrate a form of dendritic bundling that is input and output specific and may play an important role in the regulation of thalamic inputs to the cingulate cortex.

Animals↗

Position of growth cones within the retinal nerve fibre layer of fetal ferrets.

Optic axons are added to the retinal nerve fibre layer of fish along its vitreal border in a chronotopic manner. Likewise, the optic tract of all vertebrate species acquires axons preferentially along the superficial surface of the pathway. We have examined the developing retina of fetal ferrets (Mustela putorius furo) aged between embryonic day 27 (E27) and E34 to see whether a similar segregation of growth cones is apparent within the mammalian retinal nerve fibre layer. The distributions of growth cone, "wrist" (thick trailing portion of the growth cone), axonal, and glial profiles were determined from electron micrographs, and expressed as a percentage of neural profiles for several retinal locations. The retinal nerve fibre layer of fetal ferrets contains radially elongated bundles of fibres composed of axonal, wrist, and growth cone profiles. Glial processes of varying density divide the adjacent bundles, occasionally subdividing them in the plane of the retina, and give rise to endfeet lining the basal lamina and separating the optic axons from the latter. Growth cones within the developing fibre layer represented about 2.4% of profiles at E28, while at later developmental stages (E34), this value fell to about 0.6%. During this period of axonal outgrowth, growth cones were not preferentially segregated toward the vitreal basal lamina or the glial endfeet within the nerve fibre layer. Rather, they were found scattered throughout the axon bundles of the fibre layer. While there were differences in the proportion of immature profiles found within the vitreal half compared to the scleral half of the fibre layer, such that more growth cones and wrists were found vitreally, there was no clear accumulation of them in association with features of the vitreal margin. The present results show that young and old optic axons course together throughout the depth of the nerve fibre layer. A chronotopic mode of pathway genesis such as seen in the optic fibre layer of fish or in the optic tract of mammals is not present in the nerve fibre layer of ferrets. Differences in growth cone behaviour in the optic fibre layer and tract indicate that the mechanisms governing pathway formation differ along its course.

Animals↗

Interlaminar connections of the superior colliculus in the tree shrew. I. The superficial gray layer.

One of the most persistent problems in the study of the superior colliculus is the relationship between its superficial and deep layers. The superficial tier of layers is considered to be visuosensory in function, whereas the deep tier is multisensory and has premotor functions. This fundamental distinction is the primary basis for the view that a visually triggered shift in the direction of gaze depends on the transfer of information from sensory cells in the superficial tier to premotor cells in the deep tier. The goal of the present experiments was to examine the interlaminar projections of the superficial gray layer in the tree shrew Tupaia belangeri. We used biocytin as the marker for tracing the pathways. The tree shrew was chosen because its large and distinctly laminated superior colliculus facilitates the task of examining connections between the layers. Biocytin was used because of its sensitivity and because it allowed us to place very small injections restricted entirely to the superficial gray layer. The results demonstrated that a prominent pathway originates in the superficial gray layer and terminates in stratum opticum. In comparison, the projection from the superficial gray layer to the layers beneath stratum opticum is extremely sparse. The pathway from the superficial gray layer to stratum opticum has a columnar distribution, extending about 100 microns rostrally and caudally from the center of the injection site. There were no signs of more remote intracollicular connections, nor of patches or bands of terminals. The biocytin injection sites also labeled pathways to nuclei as distant from the superior colliculus as the diencephalon, including the dorsal and ventral lateral geniculate bodies, and the pulvinar. The results suggest that stratum opticum may serve as a link between the superficial gray layer and the deeper layers.

Animals↗

Ultrastructural aspects of the development of the hyaline layer and extracellular matrix lining the gastrointestinal tract in embryos and larvae of the starfish Pisaster ochraceus preserved by freeze substitution.

Embryos and larvae of the starfish Pisaster ochraceus are surrounded by a complex extracellular matrix (ECM) layer called the hyaline layer (HL). A similar but less well-organized ECM layer lines some regions of the larval gut. Examination of material preserved by freeze substitution shows that the HL consists of a coarse outer meshwork, a boundary layer, a supporting layer, which is divided into three sublayers, H1, H2, and H3, and an intervillus layer. The development of the HL has been studied in material preserved by freeze substitution. Development begins at fertilization when exocytosis of the cortical granules releases ECM into the perivitelline space and elevates the fertilization membrane. Shortly after, plaques of dense material with attached fibers are present on the outer surface of the egg plasmalemma. Following this, these plaques and fibers are associated with the tips of short microvilli, suggesting that they may induce microvillus formation. Next, the tips of some of the microvilli are joined by short regions of the H1 sublayer. Some of these H1 regions have short segments of boundary layer material associated with their outer surfaces while others are naked. Just prior to hatching, the H1 and boundary layers completely surround the embryo, separating the developing coarse meshwork and intervillus layers. Short segments of the H2 and H3 sublayers are also present. Posthatching, the microvilli and all HL layers increase in thickness and density, particularly the H2, boundary, and coarse outer meshwork layers. The results suggest a sequential organization of HL components from ECM that is secreted into the perivitelline space.

Animals↗

Ovarian receptors for insulin and insulin-like growth factor I (IGF-I) and effects of IGF-I on steroid production by isolated follicular layers of the preovulatory coho salmon ovarian follicle.

In this study, receptors for insulin and insulin-like growth factor I (IGF-I) in isolated theca-interstitial layers and granulosa cells of the coho salmon preovulatory ovary were characterized, and the effects of IGF-I on ovarian steroidogenesis were examined. Specific receptors for insulin and IGF-I were found in granulosa and theca-interstitial layers. In both follicular layers, IGF-I receptors were greater in number and higher in affinity than insulin receptors. The effects of IGF-I on in vitro production of testosterone (T) and 17 alpha-hydroxyprogesterone (17OH-P) by theca-interstitial layers and of 17 beta-estradiol (E2) and 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (17,20 beta-P) by granulosa cell layers were evaluated during the preovulatory period. Both human and salmon recombinant IGF-I inhibited the basal and GTH II-stimulated T and 17OH-P production by theca-interstitial layers throughout the preovulatory period. In contrast, IGF-I stimulated the production of both E2 and 17,20 beta-P by granulosa cell layers prior to germinal vesicle breakdown (GVBD) but only stimulated the production of 17,20 beta-P by granulosa cell layers after GVBD. The inhibitory effects of IGF-I on steroid production by the theca-interstitial layer and the opposite stimulatory effects on steroid production by the granulosa cell layer, coupled by the presence of specific IGF-I receptors in both follicular layers, suggest that IGF-I may play a role in the regulation of steroidogenesis in the preovulatory coho salmon ovary.

Animals↗

Characterization of surface layer proteins from different Clostridium difficile clinical isolates.

In a previous study we suggested that two surface proteins of a Clostridium difficile strain were involved in the formation of a regularly assembled surface layer (S-layer) external to the cell wall. In the present paper six C. difficile strains isolated from cases and healthy carriers were studied. By using freeze-etching and negative staining techniques two superimposed structurally different lattices were detected on the cell surface of the different C. difficile strains. In each strain, the outer S-layer lattice was arranged in a square symmetry and the inner S-layer lattice in hexagonal symmetry. The S-layer proteins from the different strains were isolated and characterized. Each strain showed two distinct S-layer glycoproteins ranging in molecular mass 36-56 kDa. Antigenic cross-reactivity among the S-layer proteins of higher molecular masses extracted from each strain was demonstrated whereas no antigenic relationship was observed among the different S-layer proteins of lower molecular masses. N-terminal sequence analysis showed the presence of common structural motifs conserved among the high S-layer proteins as well as among the low S-layer proteins. These data indicate that the presence of S-layer on C. difficile strains is common and that its glycoprotein subunits show a certain degree of heterogeneity.

Adult↗

Zinc-positive afferents to the rat septum originate from distinct subpopulations of zinc-containing neurons in the hippocampal areas and layers. A combined fluoro-gold tracing and histochemical study.

The purpose of the present study was to examine whether zinc-positive and zinc-negative hippocampal neurons in rats differed with respect to their projections to the septum. By combining retrograde axonal transport of the fluorescent tracer Fluoro-Gold with histochemical demonstration of zinc selenide complexes in zinc-containing neurons after intraperitoneal injection of sodium selenite, we were able to visualize the distribution of retrogradely Fluoro-Gold labeled neurons and zinc-containing neurons in the same sections. After unilateral injection of Fluoro-Gold into the rat septum a few retrogradely labeled cells were observed in layer IV of the ipsilateral medial entorhinal area, and numerous labeled cells were observed mainly in the superficial layers of the ipsilateral subicular areas and throughout the CA1 and CA3 pyramidal cell layers, as well as in the contralateral CA3 pyramidal cell layer. Zinc-containing neurons were observed in layers IV-VI of the medial entorhinal area, layers II and III of the parasubiculum, layers II, III and V of presubiculum, and in the superficial CA1 and deep CA3 pyramidal cell layers. Cells double-labeled with Fluoro-Gold and zinc selenide complexes were primarily located in distal (relative to the area dentata) parts of the superficial CA1 pyramidal cell layer and distal parts of the deep CA3 pyramidal cell layer and in layers II and III of presubiculum. Only a very few double-labeled cells were seen in the contralateral CA3. The result demonstrates that the hippocampo-septal projection of rats is a mixture of zinc-positive and zinc-negative fibers.(ABSTRACT TRUNCATED AT 250 WORDS)

Afferent Pathways↗