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Stoichiometric chrysotile single crystals have been synthesized as a unique phase by hydrothermal reaction under controlled conditions; the synthesized monocrystals show a cylinder-in-cylinder morphology and can be used as a reference sample with definite chemical composition to investigate the factors responsible of the chrysotile cytotoxicities and carcinogenicities.
Schwann cells were isolated and cultured from the twitcher mouse, an authentic model of human globoid cell leukodystrophy. We observed some difference in morphology and initial cell growth between the cells from suckling and adult mice. Initial growth was slower in homozygous and heterozygous twitcher (twi/twi; twi/+) cells than in normal cells (+/+) when cells were cultured from suckling mice. On the other hand, cell growth was initially accelerated in cells from adult twi/twi mice compared with adult twi/+ or +/+ animals. The yield of Schwann cells was much less from adults. In the cells from adult twi/twi, the perikaryon was swollen, and typical inclusions were easily recognized ultrastructurally, in particular after 14 days in vitro. However, only a few globoid cell leukodystrophy inclusions were detected in the cells from suckling twi/twi mice even after 28 days in vitro. The results indicate that Schwann cell pathology can be studied in twi/twi using an in vitro system and that twi/twi Schwann cells from suckling and adult mice have some differences in morphology as well as in the degree of initial growth.
There is a tendency in St. Petersburg since 1990s to an increase of dysentery morbidity and lethality mainly caused by Flexner Shigella 2a. Morphology of the disease in adults is given on the material of 286 necropsies in 1993-1994 in the S. P. Botkin Infectious Hospital. Males (mean age 50 years) and among them chronic alcoholics prevailed; females (mean age 70 years) were frequently with a grave cardiovascular pathology. Advanced inflammation (fibrinous, ulcerative and hemorrhagic) of all colon and distal part of the small bowel was a common finding. Hemorrhagic infarctions of the ileum which were not described so far were observed in 10% of the cases.
We have investigated biochemical and ultrastructural aspects of the nuclear matrix during the naturally synchronous cell cycle of Physarum polycephalum. The morphology of the in situ nuclear matrix exhibited significant cell cycle changes as revealed by electron microscopic examination, especially during the progression of nuclei through mitosis and S-phase. In mitosis the interchromatin matrix was found to be retracted to the nuclear periphery; during S-phase this interchromatin matrix gradually resembled, concomitant with the reconstruction of a nucleolar remnant structure. During the G2-period no significant changes in matrix morphology were observed. The pattern of nuclear matrix proteins was invariant during the cell cycle; no cycle phase-specific proteins could be detected. In vivo labelling of plasmodia with [35S]methionine/cysteine showed that only a few proteins are synthesized and assembled into nuclear matrix structures in a cell cycle-dependent way; the majority of proteins were synthesized almost continuously. This was also shown for nuclear lamins homologues. In contrast to bulk nuclear histones, those histones that remain tightly bound to the nuclear matrix were synthesized and assembled into nuclear structures in the very first hour of S-phase; assembly was terminated in mid-S-phase, indicating that nuclear matrix-bound chromatin is replicated early in S-phase. Comparison of the acetylation pattern of matrix-bound histone H4 with bulk nuclear H4 revealed a largely elevated acetate content of matrix H4. The percentage of acetylated subspecies was entirely different from that in bulk nuclear H4, indicating that matrix-associated histones represent a subpopulation of nuclear histones with distinct properties, reflecting specific structural requirements of matrix-attached chromatin.
Two types of tegmental pedunculopontine nucleus neurons have been reported previously based on their electrophysiological characteristics: type I neurons were characterized by low-threshold Ca spikes and type II neurons displayed a transient outward current. This report describes the membrane properties, synaptic inputs, morphologies and axonal projections of two subgroups of type II neurons examined in an in vitro slice preparation. Type II neurons were divided into two groups based on their spike durations: short-duration neurons with an action potential duration of 0.7-1.5 ms and long-duration neurons with an action potential duration of 1.6- 2.9 ms. Choline acetyltransferase immunohistochemistry combined with biocytin labeling indicated that 56% of short-duration neurons and 61% of long-duration neurons were immunopositive for choline acetyltransferase. Short-duration neurons had a high input resistance and the capacity to discharge with high frequency. By contrast, long-duration neurons had a low input resistance and low firing frequency and upon current injection displayed an accommodation (spike-frequency adaptation) before reaching a steady firing frequency. Microstimulation of the substantia nigra pars compacta evoked antidromic responses in both short-duration neurons (n=5/14, 36%) and long-duration neurons (n=20/39. 51%). Stimulations of the subthalamic nucleus and the substantia nigra pars reticulata induced in these neurons excitatory and inhibitory postsynaptic potentials, respectively. Short-duration neurons were dispersed equally throughout the extent of the tegmental pedunculopontine nucleus area, while long-duration neurons were located more in the rostral tegmental pedunculopontine nucleus. Short-duration neurons were small with two to four thin primary dendrites. Long-duration neurons were medium to large with three to six thick primary dendrites. Cell size was positively correlated with spike duration and axonal conduction velocity, but negatively with input resistance and spontaneous firing frequency. Both groups of neurons had ascending (toward thalamus, pretectal areas and tectum) and descending (toward pontomedullary reticular formation) axons in addition to nigropetal axons. Ascending axons were observed in 75% (6/8) of short-duration neurons and in 45% (15/33) of long-duration neurons, while nigropetal axons were observed in 50% (4/8) of short-duration neurons and in 76% (25/33) of long-duration neurons. These results suggest that the tegmental pedunculopontine nucleus cholinergic projection system is composed of heterogeneous populations of neurons in terms of electrophysiological and morphological characteristics as well as their distribution patterns in the nucleus.
The present and accompanying (Casini, G., and N.C. Brecha, J. Comp. Neurol. 326:302-313, 1992) papers investigate the postnatal development of tyrosine hydroxylase (TH)-immunoreactive (IR) amacrine cells in the rabbit retina. This study is focused on a detailed analysis of the patterns of cellular growth and differentiation of TH-IR amacrine cells, which serve as a model to gain insights into the mechanisms underlying developmental changes associated with the maturation of amacrine cells. Faintly staining TH-IR neurons are present in the proximal inner nuclear layer of newborn retinas. They are characterized by a large nucleus and usually a single primary process lacking varicosities. At postnatal day (PND) 6, TH-IR processes display more complex morphological characteristics, including a few varicosities, and second- and third-order ramifications. Growth cones are often seen. At PNDs 10 and 12 (eye opening), TH-IR cells have general morphological characteristics similar to adult TH-IR amacrines. They display 2-5 primary processes, which start forming a complex network of fibers in lamina 1 of the inner plexiform layer (IPL). TH-IR processes are also present in lamina 3 and rarely in lamina 5 of the IPL. Many fibers ending in growth cones are observed. In addition, very rare, thin TH-IR fibers are present in the outer plexiform layer. At PND 19, TH-IR fibers form a complex, dense network in lamina 1 of the IPL, and loose networks in laminae 3 and 5. Growth cones are not observed at this age. At PND 26, a few "ring-like" structures formed by TH-IR fibers in lamina 1 of the IPL are observed for the first time. In adult retinas, the "ring-like" structures are more numerous than at PND 26. A second, rare type of TH-IR cell ("type B") is encountered in all retinal regions beginning at PND 10. These cells are characterized by weak immunostaining and a small soma size. The present findings show that a significant differentiation of TH-IR neurons occurs during the first 10-12 PNDs. Eye opening is an important period for the maturation of TH-IR amacrines and, more generally, for the maturation of the IPL.
Subcutaneous tumours were induced in castrated golden Syrian hamsters by 7,12 dimethylbenz[a]anthracene (DMBA), an agent known to produce papillomas and carcinomas. The morphological characteristics of the cellular and extracellular constituents of the chemically-induced tumours were indicative of melanoma. Tumours were induced by three injections of DMBA into the jugular vein over a 3 month period. Dermal tumour development within the dorsal integument and groin region ultimately projected into the epidermis and occurred during the 3 month period subsequent to the last DMBA injection. Suspect melanoma tumours were excised and processed for light microscopic (LM) and transmission electron microscopic (TEM) studies. Histochemical staining methods facilitated the characterization of the differentiated tumour components in this hamster melanoma model. The model presented could allow observations from initial melanoma transformation events through advanced stages of metastasis within a window of 7 months.
Disks consisting of macroporous nickel-titanium alloy (NiTi, Nitinol, Actipore) are used as implants in clinical surgery, e.g. for fixation of spinal dysfunctions. The morphological properties were studied by scanning electron microscopy (SEM) and by synchrotron radiation-based microtomography (SRmuCT). The composition was studied by X-ray diffractometry (XRD), differential scanning calorimetry (DSC), and energy-dispersive X-ray spectroscopy (EDX). The mechanical properties were studied with temperature-dependent dynamical mechanical analysis (DMA). Studies on the biocompatibility were performed by co-incubation of porous NiTi samples with isolated peripheral blood leukocyte fractions (polymorphonuclear neutrophil granulocytes, PMN; peripheral blood mononuclear leukocytes, PBMC) in comparison with control cultures without NiTi samples. The cell adherence to the NiTi surface was analyzed by fluorescence microscopy and scanning electron microscopy. The activation of adherent leukocytes was analyzed by measurement of the released cytokines using enzyme-linked immunosorbent assay (ELISA). The cytokine response of PMN (analyzed by the release of IL-1ra and IL-8) was not significantly different between cell cultures with or without NiTi. There was a significant increase in the release of IL-1ra (p<0.001), IL-6 (p<0.05), and IL-8 (p<0.05) from PBMC in the presence of NiTi samples. In contrast, the release of TNF-alpha by PBMC was not significantly elevated in the presence of NiTi. IL-2 was released from PBMC only in the range of the lower detection limit in all cell cultures. The material, clearly macroporous with an interconnecting porosity, consists of NiTi (martensite; monoclinic, and austenite; cubic) with small impurities of NiTi2 and possibly NiC(x). The material is not superelastic upon manual compression and shows a good biocompatibility.
The phylogenetic relationships of Pyricularia species and species from related genera were established from sequences of the internal transcribed spacer ribosomal RNA gene. Phylogenetic analysis disclosed a consistent correlation with spore morphology. Most Pyricularia species studied, and two species of Dactylaria that have obpyriform conidia, fell within the Magnaporthaceae cluster with high bootstrap support. Pyricularia variabilis was more related to Dactylaria, Tumularia or Ochroconis species than to the Magnaporthaceae. Dactylaria and species of Nakataea, Ochroconis, Pyriculariopsis and Tumularia were distinct from the Magnaporthaceae, and the genus Dactylaria is polyphyletic. The combination of morphological and molecular characters, such as spore morphology and ITS ribosomal DNA sequences data, suggested that conidial shape could be a primary character to distinguish Pyricularia from related genera.
Peptide nucleic acid amphiphiles (PNAA) are a promising set of materials for sequence-specific separation of nucleic acids from complex mixtures. To implement PNAA in micellar separations, the morphology and size of PNAA micelles in the presence and absence of a sodium dodecyl sulfate (SDS) cosurfactant have been studied by small-angle X-ray scattering and dynamic light scattering. We find that a 6-mer PNAA with a 12-carbon n-alkane tail forms ellipsoidal micelles (a = 5.15 nm; b = 3.20 nm) above its critical micelle concentration (CMC) of 110.9 microM. On addition of a stoichiometric amount of complementary DNA, PNAA hybridizes to DNA, suppressing the formation of PNAA micelles. At a ratio of 19:1 SDS/PNAA (total concentration = 20 mM), spherical micelles are formed with outer radius Rs = 2.67 nm, slightly larger than spherical micelles of pure SDS. Capillary electrophoresis studies show that PNAA/DNA duplexes do not comicellize with SDS micelles. No such effects are observed using noncomplementary DNA. The shape and size of the PNAA micelles is also verified by dynamic light scattering (DLS) studies. These results provide an interesting case study with competing electrostatic, hydrophobic, and hydrogen-bonding interactions in micellar systems and make possible the use of PNAA in micellar separations of DNA oligomers.
A group of 32 annatto genotypes collected in five Venezuelan regions (Oriente, Centro, Llanos, Andes and Amazonas) and in Brazil were used for morphological studies. The fruit variables with the greatest discriminatory power in the formation of groups were capsule size, spinosity and seed size. On the other hand, an association group among the variables spinosity, spine length, dehiscence and apex shape were formed, also a proportional association between capsule and seed size, and between dehiscent capsule and brown coloured seeds. Additionally, in order to discriminate morphological variables behaviour in respond to electrophoretic variables, a group of protein and isozyme bands associated with fruit characteristics were established. Therefore, a classification system of this species was possible using morphological studies of the capsules, even though a determined association relating morphological and molecular patterns was not found.
The effects of self-etching adhesives on primary teeth were evaluated. Reports in the literature suggest differences between the first and second dentition regarding the composition and the morphology and, therefore, a possible difference in the performance of dental adhesives. The purpose of this study was to evaluate the effects of self-etching adhesives on primary dentin. Eight dentin disks were obtained from the occlusal surface of primary molars. The disks were divided between four dental adhesives [Prompt L-PoP (LP), Clearfil SE BOND (SE), Etch&Prime 3.0 (EP) and Prime&Bond NT (PB) + H(3)PO(4) (Control) and "restored" with a composite (Pertac II)]. After sectioning, fixation and HMDS drying, specimens were polished and Field Emission SEM examinations were carried out. Clearly visible hybrid layer formation was found for PB, LP and EP. An undoubtedly detectable interdiffusion zone was not evident after the use of SE. A clearly visible adhesive layer was recognizable for PB, EP and SE, but not continuously detectable for LP. Debonded regions were observed for all systems evaluated, but distinct differences in the failure mode were detected. The evaluated dental adhesives did not generate completely sealed interfaces between the composite resin and the dentin of primary teeth in vitro.
Ulcerations and intraplaque hemorrhages are thought to play an important role in the development of neurological deficits in carotid artery stenoses. To assess the ability of duplex scanning to predict plaque morphology we compared different sonographic criteria (plaque border, plaque density, plaque structure) with the morphological findings in 169 consecutive carotid endarterectomies (144 cases with diameter reduction greater than 50%). Two percent of the sonograms were inadequate, and 20% showed poor image quality. Regular plaque borders revealed a smooth or at most minimally ulcerated surface in 92%. Grossly ulcerated stenoses, however, were only found in 27% of the irregular plaques. Moreover, the plaque border was nonvisible in 35% of all cases. Simple, fibro-atheromatous plaques were found to be echogenic in 72% and echolucency was present in 80% of the stenoses with relevant intraplaque hemorrhage. In conclusion, duplex scanning proved to be capable of detecting smooth, fibro-atheromatous stenoses with high accuracy. Ulcerations, however, cannot be predicted reliably, and intraplaque hemorrhage cannot be differentiated from atheromatous debris.
Selective removal of the basal forebrain cholinergic neurons by the immunotoxin 192 immunoglobulin G-saporin has offered a new powerful tool for the study of the relationships between cholinergic dysfunction and cognitive impairments. In the present study the morphological and functional consequences of selective lesions of the basal forebrain cholinergic system during early postnatal development have been investigated following bilateral intraventricular injections of 192 immunoglobulin G-saporin to immature (four-day-old) rats. Administration of increasing doses (0.2-0.8 microgram) of the immunotoxin produced dose-dependent loss of cholinergic neurons in the septal/diagonal band area (up to 72-86%) and in the nucleus basalis magnocellularis (up to 91-93%), paralleled by marked reductions in choline acetyltransferase activity in the hippocampus and several cortical regions (73-84%). The parvalbumin-positive neurons in the septal/diagonal band area and the calbindin-positive Purkinje cells in the cerebellum were unaffected at all dose levels. Brain dopamine or noradrenaline levels were unaffected or increased by the immunotoxin treatment. At the optimal dose, 0.4 microgram, the toxin conjugate produced maximal cholinergic depletion without significant mortality. Higher doses (0.8, 1.2 and 1.6 micrograms) of toxin, on the other hand, proved to be lethal for most or all of the injected animals. When tested at three and eight months after the optimal dose, in spite of persisting cholinergic depletion, the noenatally lesioned animals showed no impairment in the water maze task or in locomotor activity and exploration as compared to normal controls, probably reflecting partial sparing of the cholinergic neurons by the neonatal immunotoxic lesion (above all in the vertical and horizontal limbs of the diagonal band area), and/or a greater degree of plasticity in the developing as compared to the mature cholinergic system. The place navigational performance of the neonatally lesioned animals in the water maze task was abolished by central muscarinic cholinergic receptor blockade (by atropine) or by a second immunotoxic lesion, which eliminated virtually all residual cholinergic neurons in the septal/diagonal band area and the nucleus basalis. Administration of 192 immunoglobulin G-saporin to similarly trained, but previously normal adult rats, produced similar cholinergic depletions but much less severe place navigation deficits, suggesting that preoperative training on the task may reduce the functional consequences of a subsequent cholinergic lesion. The results thus support the view that the basal forebrain cholinergic system may be implicated in the acquisition rather than retention of spatial memory in the water maze task.
The presence of Aedes albopictus (Skuse) was recently confirmed for the first time in northwestern Greece. This location is within the distribution range of a morphologically similar species, Aedes cretinus Edwards, and is a potentially favorable region for the reintroduction of Aedes aegypti (L.). It was thus compelling to use methods in addition to morphology-based keys to correctly identify specimens badly damaged, rubbed, or otherwise altered in their external characteristics. It was decided to use molecular techniques as a novel and reliable method for differentiating the three Stegomyia species. The nuclear internal transcribed spacer 2 (ITS2) fragments from morphologically identified Ae. albopictus and Ae. cretinus specimens were amplified, and their sequences were compared with those in GenBank for Ae. albopictus, Ae. cretinus, and Ae. aegypti. Also, mitochondrial cytochrome oxidase I (COI) fragments were amplified for Ae. albopictus and Ae. cretinus (so far not available in GenBank) and compared with Ae. aegypti fragments. ITS2 and COI sequences generated in our study were deposited in GenBank and could be useful in future studies of mosquitoes by other research workers.
In addition to a modulatory function, somatostatin (SS) is likely to exert a morphogenetic and/or trophic role in the developing nervous system. In this study, a mouse monoclonal antibody directed to SS was used to investigate the posthatching development of SS-immunoreactivity (SS-ir) in the pigeon retina to provide a basis for a better understanding of the role of this peptide in retinal maturation. In the adult, SS-ir was observed in amacrine cells located in the inner nuclear layer (INL) of the entire retina. Two cell types were recognized according to their morphology. They showed a differential density distribution. Cell type indicated as "adult 1" (AD1) was characterized by pear-shaped cell bodies with single primary processes directed to the inner plexiform layer (IPL) and was mostly present in the red field. In contrast, cell type indicated as "adult 2" (AD2) was characterized by round-shaped somata with 1-3 primary processes and was highly represented in the fovea and the dorsal periphery. Posthatching maturation of the pigeon retina was characterized by drastic changes in the pattern of SS-ir. Over the first days posthatching, SS-ir was observed in sparsely distributed somata mostly located in the ganglion cell layer (GCL). This cell type indicated as "hatch" (H) was characterized by dense granular staining and became extremely rare at 7 days. Over the same period, growing SS-positive axons displaying enlarged growth cones were found in the optic tract (TrO). These observations suggest the possibility that ganglion cells transiently expressing SS are present at early stages of posthatching development. Of the two types of SS-containing cells observed in the adult, the first to be recognized morphologically was cell type AD1 which appeared at 2 days after hatching in the INL. These cells were virtually adult-like in morphology by 7 days. In contrast, cell type AD2 was not apparent until 7 days posthatching. The density (defined as number of cells/mm2 of retinal tissue) and the total number of SS-containing cells changed during posthatching maturation. In particular, the adult number of cell type AD1 was reached at about 10 days, while the number of cell type AD2 was reached at about 3 weeks posthatching. At this stage, both cell types also displayed their mature density distribution. The present findings suggest a temporal relationship between the maturation of SS-ir and developmental events which include the onset of light-driven activity and the maturation of retinal acuity.(ABSTRACT TRUNCATED AT 400 WORDS)
Classically, primordial follicles have flattened pre-granulosa cells that become cuboidal and divide during follicular activation. This change in shape is thus an index of activation. To examine this and follicular morphology in cattle, ovaries were processed for light and electron microscopy (n = 21). In single sections (5 microns thick), primordial follicles had a mean ratio of maximum to minimum diameters of 1.33 +/- 0.18 (mean +/- SD; n = 317, randomly selected), indicating that they had a prolate rather than a spherical shape. The prolate shape was attributable to clustering of granulosa cells at two opposite poles on the long axis of each follicle; and in each histological section, 82.5% of primordial follicles (n = 317) had at least one cuboidal granulosa cell. More accurate measurements of primordial follicles indicated that they had three dimensions measured as length (45.4 +/- 2.4 microns), breadth (26.8 +/- 1.5 microns), and depth (< 30.4 +/- 1.4 microns) (mean +/- SEM; n = 12, constructed from serial sections); a surface area of 0.0032 mm2; and 24 +/- 2 granulosa cells. The ultrastructural morphology of the cells of the bovine primordial follicle (n = 31 examined by electron microscopy) was similar to that of other species. The ovarian cortex was composed of discrete zones. The zones containing primordial follicles were substantially avascular, as observed by localizing von Willebrand factor, and rich in collagen fibrils. In conclusion, since primordial follicles of bovine ovaries contain some cuboidal pre-granulosa cells, changes in shape cannot be used with certainty as a marker of activation of bovine follicles.
Phosphate-activated glutaminase (PAG) is the major enzyme involved in the synthesis of the excitatory neurotransmitter glutamate in cortical neurons of the mammalian cerebral cortex. In this study, the distribution and morphology of glutamatergic neurons in cat visual cortex was monitored through immunocytochemistry for PAG. We first determined the specificity of the anti-rat brain PAG polyclonal antibody for cat brain PAG. We then examined the laminar expression profile and the phenotype of PAG-immunopositive neurons in area 17 and 18 of cat visual cortex. Neuronal cell bodies with moderate to intense PAG immunoreactivity were distributed throughout cortical layers II-VI and near the border with the white matter of both visual areas. The largest and most intensely labeled cells were mainly restricted to cortical layers III and V. Careful examination of the typology of PAG-immunoreactive cells based on the size and shape of the cell body together with the dendritic pattern indicated that the vast majority of these cells were pyramidal neurons. However, PAG immunoreactivity was also observed in a paucity of non-pyramidal neurons in cortical layers IV and VI of both visual areas. To further characterize the PAG-immunopositive neuronal population we performed double-stainings between PAG and three calcium-binding proteins, parvalbumin, calbindin and calretinin, to determine whether GABAergic non-pyramidal cells can express PAG, and neurofilament protein, a marker for a subset of pyramidal neurons in mammalian neocortex. We here present PAG as a neurochemical marker to map excitatory cortical neurons that use the amino acid glutamate as their neurotransmitter in cat visual cortex.