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  • 1990-1994  (72)
  • 1975-1979
  • 1920-1924
  • 1890-1899
  • 1870-1879
  • 1993  (72)
  • Electron microscopy
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 71 (1993), S. 452-460 
    ISSN: 1432-1440
    Keywords: Electron microscopy ; Alveolitis ; Pulmonary fibrosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Lung specimens of 21 patients with diffuse interstitial lung disease were examined. The present ultrastructural study outlines the topography and distribution of inflammatory changes in the interstitium, endothelium, and in pneumocytes and phagocytes. Alveolitis is characterized by marked regenerative activity of type II pneumocytes (cuboid metaplasia), intraluminal macrophage accumulation, endothelial swelling, multilamination of the endothelial basement membrane, pericapillary edema, and primarily by cellular infiltrates in the interstitial space. The most prominent feature of the interstitium in pulmonary fibrosis is the lack of immunoinflammatory cells. In some areas there is a marked absence of alveolar lumen while only a small number of macrophages are present in the remaining alveolar lumen. Most of the capillaries in the fibrous septum have been destroyed. Ultrastructural studies of lung biopsies in patients with diffuse interstitial lung disease allow the differentiation between alveolitis and pulmonary fibrosis and thus contribute to a therapeutic decision.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Child's nervous system 9 (1993), S. 246-249 
    ISSN: 1433-0350
    Keywords: Infantile myofibromatosis, solitary type ; Intracranial myofibromatosis ; Immunohistochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract An unusual case of infantile myofibromatosis of the solitary type occurring in an intracranial location in a 48-day-old female infant is presented. To our knowledge, there are no other descriptions in the literature of infantile myofibromatosis with exclusively intracranial involvement. The immunohistochemical and electron microscopic findings confirm the myofibroblastic origin of the proliferation.
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  • 3
    ISSN: 1432-2285
    Keywords: Picea abies (L.) Karst ; Freezing injury ; Acid rain ; Carbohydrate histochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The cellular structures of acid rain-irrigated needles of several provenances of Norway spruce (Picea abies L. Karst) seedlings were studied after winter experimental freezing. Frost injuries and recovery were characterized by visual damage scoring and classification of mesophyll cell alterations, also using histochemical methods for carbohydrate fluorescent staining. The treatment with-30° C during the late dormancy period was sufficient to cause significant injuries and intracellular degradation in the tissues of the green needles. The most affected seedlings in terms of visual injury scoring were found among those treated with clean water or at pH 3, while freezing injury, defined as an occlusion of phenolic substances in the central vacuole of the mesophyll cells, was most abundant in the needles from spruces irrigated either with clean water or at pH 4 or pH 3. Electron microscopy revealed the details of the injury, e. g. thinning out of the cytoplasm and chloroplast stroma, darkening of the chloroplasts and eventually swelling of the chloroplasts and protoplast. PAS and ConA reactions in the needle tissue revealed intense starch accumulation in the mesophyll and transfusion tissues as early as in March, with a tendency to increase, especially in the untreated needles during the recovery period. Plasma membrane disturbances were indicated by histochemical identification of callose deposits in the mesophyll cell walls, these being most abundant in the acid rain-treated needles. All these findings suggest that freezing at −30° C was more deleterious to the seedlings pretreated with acid or clean water than to those not given additional irrigation.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Trees 8 (1993), S. 23-30 
    ISSN: 1432-2285
    Keywords: Wound responses ; Hardwoods ; Xylem parenchyma ; Suberization ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Wound responses of xylem parenchyma by suberization were investigated in some hardwoods by light and electron microscopy. Suberized ray and axial parenchyma cells form a distinct boundary around the wound in all investigated species. Vessels and fibres within and close behind the suberized area appeared more or less occluded; vessels in Fagus, Quercus, and Populus contained suberized tyloses, those in Betula and Tilia contained amorphous and fibrillar deposits. A common mechanism for suberin deposition in the parenchyma cells became evident. Cisternae of the endoplasmic reticulum were apparently involved in suberization. Suberin compounds are extruded by cytoplasmic vesicles, which fused with the plasma membrane, in order to release their content. The suberin layer exhibited the typical lamellated structure; cytoplasmic continuity between suberized cells by plasmodesmata was maintained through the suberin layer. Fagus revealed the most intense suberized area as compared with the other species. Within the reaction zone of Fagus and Quercus, some individual ray and axial parenchyma cells exhibited a subdivision into 2 or 3 compartments prior to suberization. Subdivision was achieved by the formation of a primary wall-like layer. Subsequently, the compartments became individually suberized. Wounding during winter did not induce suberization. Also, samples wounded and kept under water during the vegetation period showed no response. The role of suberization in the effectivity of wound-associated compartmentalization is discussed.
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  • 5
    ISSN: 1432-2307
    Keywords: Chromomycosis ; Skin ; Granuloma ; Immunohistochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The cell-mediated immune reaction was studied in the cutaneous lesion of chromomycosis, using monoclonal antibodies against polymorphonuclear neutrophils, macrophage and lymphocyte subsets, endothelial and fibroblast cells. In addition, immunostaining of the main degradative enzymes (neutrophil elastase and interstitial collagenase) and certain important cytokines (transforming growth factor-β, tumour necrosis factor-α and interferon-γ) suggested an explanation for the granulomatous reaction and the associated tissue remodelling. The distribution pattern of neutrophils and macrophage subsets, observed by computer-aided image analysis, suggests that the in situ persistence of fungi is the main pathological factor.
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  • 6
    ISSN: 1432-2307
    Keywords: Solitary fibrous tumour ; Mesenchymal tumour ; Thyroid ; Immunohistochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We describe three cases of solitary fibrous tumour (SFT) arising from thyroid stroma. Grossly, the tumours were clearly delimited but only partly encapsulated. The following histomorphological growth patterns were observed: bundles of cells in storiform configuration; non-structured bundles; prevalence of fibrous matrix; highly cellular, non-structured; prevalence of loose, non-structured extracellular substance; cellular proliferation and vascular spaces in a haemangiopericytic configuration and a lipomatous component. Immunohistochemical investigation demonstrated intense, diffuse vimentin positivity and focal, less intense actin positivity in all three cases. At electron microscopy we observed a primitive cell of mesenchymal type, with cytoplasm poor in organelles and rich in filaments; this cell sometimes presented differentiation characteristics. SFT is at present the most correct term for the lesions presented here despite some morphological characteristics which differ from cases reported in the literature.
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  • 7
    ISSN: 1432-2307
    Keywords: Multiple papillary adenomas ; Type II pneumocyte ; Von Recklinghausen's disease ; Immunohistochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A case of multiple papillary adenomas of type II pneumocytes is reported. A 13-year-old boy with von Recklinghausen's disease had small nodular lesions in both lungs without symptoms. The biopsied lung contained greyish-white nodules ranging in size from 0.5 to 2 mm. Light microscopic examination revealed cuboidal to low columnar cells arranged in a papillary pattern. Elastic fibres were present in the tumour stroma. Electron microscopically, the cells had osmiophilic lamellar bodies in the cytoplasm and short microvilli along the free border. The tumour cells expressed immunoreactivity for epithelial membrane antigen and surfactant apoprotein antibodies. More than 6 years after open lung biopsy, the patient is well but small nodular shadows can still be identified.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Virchows Archiv 423 (1993), S. 469-476 
    ISSN: 1432-2307
    Keywords: Ciliary regulation ; Cytoskeleton ; Bronchial epithelium ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Observations of explanted bronchial mucosa show that ciliary function is maintained for 7 days subsequent to explantation. This finding demonstrates that non-neural mechanisms exist which regulate ciliary function. Ultrastructural and immunohistochemical studies both for light and electron microscopy were performed on human bronchial biopsy material and lung resection specimens in order to recognize the morphological substrate of this regulatory mechanism. A complex system of cytokeratin filaments and microtubules radiate through the whole cytoplasm of ciliated cells with direct contact to the nucleus, cilia, microvilli, desmosomes and to the apical terminal adhesive complex. Between the basal bodies and the apical terminal adhesive complex microfilaments can be found. In the apical cytoplasm a dense filamentary network is seen in association with the adhesive complex. These morphological findings indicate that the cytoskeleton of the bronchial epithelium plays a key role in the co-ordination of ciliary function.
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  • 9
    ISSN: 1432-2307
    Keywords: Muscle regeneration ; Myonuclei ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Muscle regeneration was studied by light and electron microscopy in a case of exercise-induced acute myoglobinuria in a young patient with carnitinepalmityl-transferase deficiency. Various stages of regeneration existed in the foci of necrosis scattered throughout apparently normal muscle. Activated satellite cells, myoblasts and myotubes were found, some of them containing myofibrils. Among the cells accumulating in the necrotic fibres, some apparently contained surviving myonuclei. In some fibres of normal size, developing myofibrils were abundant. Surviving myonuclei may be of significance in the reaction of muscle cells after injury.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Virchows Archiv 423 (1993), S. 39-43 
    ISSN: 1432-2307
    Keywords: Pseudomonas aeruginosa ; Bacterial adherence ; Bronchial mucosa ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The route of bacterial infection of the lower respiratory tract is generally one of descent subsequent to colonisation of the oral and oropharyngeal mucosa. The interaction betweenPseudomonas aeruginosa (wild type) and the bronchial epithelium was studied in bronchial mucosal probes cultured in tissue culture medium. It was possible to demonstrate that, even after loss of the mucus layer, adherence between the bacteria and the bronchial epithelium does not take place if ciliary function remains intact. Only after mechanical destruction of the bronchial epithelium, in proximity to squamous metaplasia or after loss or malfunction of the cilia of the bronchial epithelial cells was adhesion between bacteria and bronchial epithelial cells or basement membrane demonstrated by electron microscopy. After loss of the cilia following adenovirus-infection, adhesion betweenP. aeruginosa and the bronchial epithelial cells was visible. These results indicate that ciliary function must be of crucial significance in bacterial epithelial colonisation.
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  • 11
    ISSN: 1432-2307
    Keywords: Parachordoma ; Extraskeletal myxoid chondrosarcoma ; Chordoma ; Immunohistochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A case of parachordoma of the left calf in a 19-year-old Chinese female is reported. The tumour showed multinodular growth pattern and consisted of round or oval tumour cells with abundant eosinophilic cytoplasm and myxoid matrix. Tumour cells formed small nests and sometimes showed concentric arrangement. Physaliferous-like cells and undifferentiated spindle cells were occasionally observed among the cell nests. The myxoid matrix was positive for high-iron diamine stain, indicating the presence of chondroitin 4- and 6- sulphates and keratan sulphate. Ultrastructurally, well-developed rough endoplasmic reticulum, abundant intermediate filaments, microvillous cytoplasmic processes, pinocytic vesicles, and desmosome-like junctional structures were found. Tumour cells were positive for S-100 protein and vimentin, but negative for cytokeratin, epithelial membrane antigen, carcinoembryonic antigen, and desmin. These results are consistent with the definition of parachordoma as a soft tissue neoplasm consisting of cells with histology and ultrastructure similar to those of chordoma cells but with immunohistochemistry similar to that of chondroid tumour cells.
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  • 12
    ISSN: 1434-0879
    Keywords: Bladder dysfunction ; Electron microscopy ; Histology ; X-irradiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The aim of this research was to establish a small animal model for the functional and morphological study of post-irradiation bladder dysfunction. Young adult female Wistar rats were X-irradiated with single doses of 10, 15, 20 or 25 Gy. Filling cystometry was performed to assess changes in reservoir function: the volume infused to produce a rise in intravesical pressure of 5 cmH2O was calculated as an index of compliance. A biphasic reduction in this index was noted in animals receiving 15–25 Gy; the first reduction developed at about 4 weeks, and the second started at 3–4 months and persisted at 6 months. Bladder tissue was taken at this time (6 months post-irradiation) for morphological study. Histological examination demonstrated an increased mast cell density in the irradiated bladders, but was otherwise non-specific; fibrosis was discernible in only half of the 18 animals studied. Electron microscopy showed focal degeneration of smooth muscle cells, and in some areas there was selective degeneration of unmyelinated axon profiles. The biphasic reduction in the compliance index is consistent with the timing of the symptoms of the acute and late irradiation reactions reported by radiotherapy patients. Changes in axon profiles and mast cell density may be of functional significance.
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Mycopathologia 121 (1993), S. 143-147 
    ISSN: 1573-0832
    Keywords: Electron microscopy ; Farmer's lung ; Saccharopolyspora rectivirgula ; Thermoactinomyces vulgaris
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The fine structure ofThermoactinomyces vulgaris andSaccharopolyspora rectivirgula is described by transmission electron microscopy. These two bacteria are the most common microbes causing farmer's lung. The fine structure of hyphae, germination of endospores and the details of conidial wall layers ofT. vulgaris, as well as the fine structure of septate hypha and globose, polygonal conidia ofS. rectivirgula are described. The conidial wall ofT. vulgaris consisted of an inner multilayered spore coat, intermediate spore coat and outer spore coat. The findings are important for the investigations to find fragments of these bacteria in the lungs of exposed patients and experimental animals.
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  • 14
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 187 (1993), S. 131-138 
    ISSN: 1432-0568
    Keywords: Beak skin ; Quail ; Free nerve ending ; Merkel nerve ending ; Grandry corpuscle ; Herbst corpuscle ; Ruffini corpuscle ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary This study is concerned with the distribution and ultrastructure of sensory nerve endings in the beak skin of adult Japanese quail (Coturnix coturnix japonica). The following nerve endings were found: free nerve endings, clusters of dermal Merkel nerve endings, Herbst corpuscles and Ruffmi corpuscles. The latter were found only in the dermis of the tip of the upper beak. The remaining endings were present in the skin of all areas of upper and lower beak. Free nerve endings were supplied by either thin myelinated axons or unmyelinated C-fibers and were localized in the dermis close to the basal layer of the epidermis. Merkel cells formed clusters (up to 50) localized below and between the epidermal cones of the beak skin. Disc-shaped thickenings of nerve endings were squeezed between individual Merkel cells. Small Herbst corpuscles were found in the dermis close to the epidermal cones of the beak skin. Large Herbst corpuscles occurred in deep layers of the dermis. The Ruffmi corpuscles were cylindrical in shape (80 μm × 400 μm) and arranged in groups of up to ten corpuscles. Each corpuscle was surrounded by an incomplete fibrous capsule.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 85 (1993), S. 323-326 
    ISSN: 1432-0533
    Keywords: Creutzfeld-Jakob disease ; Electron microscopy ; Giant collagen plaques
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We report two cases of Creutzfeld-Jakob disease with clusters of giant collagen fibers. To our knowledge, these abnormally large collagen fibers have never been described in patients with degenerative diseases of the central nervous system. The significance of the formation of such plaque-like large collagen fibers has as yet not been elucidated. It is felt that these represent a product of the degenerative process.
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  • 16
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 86 (1993), S. 42-48 
    ISSN: 1432-0533
    Keywords: Streptozotocin diabetes ; Electron microscopy ; Giant axonopathy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The ultrastructure of peripheral sensory nerves was investigated in adult Wistar rats suffering from experimental diabetes mellitus 6 and 10 weeks after the injection of streptozotocin. Giant axons were seen in sections from the nerves of streptozotocintreated rats; some contained masses of neurofilaments, others were predominantly filled with ill-defined vesicles. At the swollen axons, the myelin sheath was thinned or absent. In other regions, large intramyelinic vacuoles were observed. A number of nerve fibers broke down completely and underwent Wallerian degeneration. This was accompanied by Schwann cell proliferation and formation of Büngner bands. Concomitantly with axonal degeneration, nerve regeneration started from intact internodes. The pathomorphology of streptozotocin diabetic neuropathy closely resembles that of some toxic distal axonopathies. This points to a common metabolic basis of giant axonopathies of different etiology.
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  • 17
    ISSN: 1432-0533
    Keywords: Pituitary adenoma ; Gigantism ; Electron microscopy ; Immunocytochemistry ; Cell culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A pituitary adenoma was transsphenoidally removed from a 4.5-year-old girl suffering from gigantism. Prior to the operation both the growth hormone (GH) and the prolactin (PRL) levels in the serum were elevated. By light microscopy the tumor appeared to be an acidophilic adenoma. Two distinct cell types, the densely granulated and the sparsely granulated cells, could be distinguished by electron microscopy. Double immunolabeling revealed the presence of GH alone in some densely granulated cells and PRL alone in some sparsely granulated cells, as well as GH and PRL co-localized in both of the morphologically distinguished cell types. Both cell types were identified in the monolayer and the suspension cultures by electron microscopy. GH and PRL concentrations in the culture media were measured by radioimmunoassay. The basal secretion of growth hormone was almost uniform during the 3-week cell culture period. GH and PRL release was significantly inhibited by bromocriptine. Our studies revealed a bimorphous and bihormonal mixed adenoma in childhood.
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  • 18
    ISSN: 1432-0533
    Keywords: Dialysis encephalopathy ; Aluminium ; Silver staining ; Laser microprobe ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Recently we described silver-staining variants for the demonstration of β/A4 amyloid and neurofibrillary tangles in senile dementia of Alzheimer type (SDAT). The same methods allowed, for the first time, the visualization of characteristical patterns and distinct morphological changes in human dialysis-associated encephalopathy. Light and electron microscopy demonstrated typical silver-stained inclusions in the cytoplasm of choroidal epithelium, glia and neurons. Performing laser microprobe mass analysis on en-bloc silver-stained semithin sections, evidence for significant amounts of aluminum was obtained within the lesions. Prominent aluminum-signals were obtained additionally in adjacent structures and nuclei of sections which were stained with toluidin-blue exclusively. Silver-stained paraffin sections of ten patients with a history of long-term hemodialysis were evaluated. The choroidal epithelium-obviously the most sensitive structure-showed black inclusions ranging from a few dots to a complete black staining of cells. Glial cells presented massive silver-stained deposits, which were restricted to the gray matter. Finally, neurons revealed numerous fine-granular black inclusions, scattered throughout the cytoplasm. Brain stem nuclei were primarily affected, but neurons within cortex, subcortical gray matter and spinal cord were also involved to various degrees; inclusions were not evident in the nucleus dentatus and the oliva inferior. Vessel-related deposits were found frequently. By electron microscopy the cytoplasm of neurons was filled either with large amounts of small electron-dense granules, or with lipofuscin granules, containing numerous irregular, non-membrane-bound inclusions. Massive electron-dense depositions were seen in the cytoplasm of choroidal epithelia and in proximity to nuclei of cortical astro- and oligodendroglia. The described neuronal changes and, in particular, alterations of choroidal epithelium and glia are completely different from characteristic plaques and tangles in SDAT.
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  • 19
    ISSN: 1432-069X
    Keywords: Human papillomavirus ; In situ hybridization ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Electron microscopic in situ hybridization (EMISH) of common warts (verrucae vulgares) of the hands was performed using a biotinylated human papillomavirus type 2a (HPV-2a) DNA probe and immunogold labelling of ultrathin sections of 2% glutaraldehyde-fixed, Lowicryl K4M-embedded tissues. It was first established that the warts contained HPV-2a DNA by light microscopic in situ hybridization. The HPV-2a probe chiefly labelled cells in the horny, granular and upper spinous layers of the epidermis, and labelling decreased towards the basal cell layer. The gold particles were located precisely on the viral particles in the nuclei of granular cells. The lower limit of detection by EMISH was found to be the keratinocytes of the third cellular layer above the basal cells. These keratinocytes showed evidence of a viral cytopathic effect, suggesting that vegetative DNA replication in infected keratinocytes occurs at least as early as this level of the epidermis.
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  • 20
    ISSN: 1432-0827
    Keywords: Chondrocytes ; High-density suspension culture ; Electron microscopy ; Matrix vesicle ; Apatite formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Chondrocyte cultures grown in centrifuge tubes with intermittent centrifugation differentiate into hypertrophic chondrocytes and form calcification. We examined chondrocytes cultured in this system electron microscopically. Rat growth-plate chondrocytes were seeded in a plastic centrifuge tube and cultured in the presence of Eagle's minimum essential medium supplemented with 10% fetal bovine serum and 50 μg of ascorbic acid per ml. Specimens were examined by using electron microscopy and selected-area electron-diffraction techniques. In the early stage of culture, a few chondrocytes were scattered and extracellular matrices were not observed. In the middle stage of the cultures, the chondrocytes resembled proliferative cells. Matrix vesicles appeared to be budding from the cell surfaces of chondrocytes and were observed sparsely in the extracellular matrices, which were well formed around the chondrocytes. Matrix vesicles increased substantially during the following cultures. In the mature stage of the cultures, crystal formation related to matrix vesicles was observed. In the 33-day cultures, several masses of calcified matrix were formed and it was confirmed to be apatite by selected-area electron diffraction analysis. The chondrocytes appeared hypertrophic during this same stage. The 56-day culture was similar to the 33-day culture. It was concluded that this culture system provides an extracellular-matrix mineralization which is produced by chondrocytes per se.
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  • 21
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 160 (1993), S. 284-287 
    ISSN: 1432-072X
    Keywords: Bacterial glucoamylase ; Clostridium thermosacharolyticum ; Cellular location ; Activity states ; Macromolecular organization ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By application of immunocytochemical techniques at the electron microscope level, glucoamylase was localized to the cell periphery in Clostridium thermosaccharolyticum during and following growth on starch, sucrose or glucose. Levels of immunolabelling were found to be relatively independent of growth substrate and of phase of growth, whereas previous studies had demonstrated strong dependence of glucoamylase activity on growth conditions; previously high levels of glucoamylase activity had been detected after growth on starch (i.e. during the stationary phase after growth) and only very low activities detected during exponential growth and following growth on glucose. The results presented demonstrate that levels of the glucoamylase protein are independent of measurable enzyme activity, and imply that the protein is constitutive. This indicates that the protein can exist in active and inactive states in the cell. By analogy with similar systems, we consider it likely that “maturation” or “activation” of newly synthesized glucoamylase occurs during (or following) transport through the cytoplasmic membrane. Electron microscopy of individual protein molecules which had been subjected to negative staining revealed that the enzyme consists of two domains of approximately equal size which are linked by a “hinge” region.
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  • 22
    ISSN: 1432-072X
    Keywords: Trichoderma reesei ; Xylanase ; Ultrastructural localization ; Immunogold labelling ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The intracellular location of the “low-molecular weight, alkaline” xylanase (XYN II) of Trichoderma reesei RUT C-30 was investigated during growth on xylan, using immunoelectron microscopy. A monoclonal antibody, produced against XYN II, was used for this purpose. The enzyme was found at the endoplasmic reticulum and in electron dense 0.2 to 0.8 μm vesicles, as well as in the vacuole, at the plasma membrane and in the fungal cell-wall. No staining occured in the cytoplasm, the mitochondria and the nucleus. No Golgi-like structures could be seen. Addition of the carboxylic ionophore monensin blocked xylanase as well as total protein secretion. The results are discussed with respect to XYN II being secreted by T. reesei via a pathway involving the endoplasmic reticulum and secretory vesicles and/or the vacuole.
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  • 23
    ISSN: 1432-072X
    Keywords: Malonomonas rubra ; Propionigenium modestum ; Malonate decarboxylase ; Methylmalonyl-CoA decarboxylase ; Biotin ; Avidin ; Electron microscopy ; High pressure freezing ; Immunolabeling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Malonate decarboxylase of Malonomonas rubra is a complex enzyme system involving cytoplasmic and membrane-bound components. One of these is a biotin-containing protein of Mr 120'000, the location of which in the cytoplasm was deduced from the following criteria: (i) If the cytoplasm was incubated with avidin and the malonate decarboxylase subsequently completed with the membrane fraction the decarboxylase activity was abolished. The corresponding incubation of the membrane with avidin, however, was without effect. (ii) Western blot analysis identified the single biotin-containing polypeptide of Mr 120'000 within the cytoplasm. (iii) Transmission electron micrographs of immuno-gold labeled M. rubra cells clearly showed the location of the biotinyl protein within the cytoplasm, whereas the same procedure with Propionigenium modestum cells indicated the location of the biotin enzyme methylmalonyl-CoA decarboxylase in the cell membrane. The biotin-containing protein of the M. rubra malonate decarboxylase enzyme system was not retained by monomeric avidin-Sepharose columns but could be isolated with this column in a catalytically inactive form in the presence of detergents. If the high binding affinity of tetrameric avidin towards biotin was reduced by destructing part of the tryptophan residues by irradiation or oxidation with periodate, the inhibition of malonate decarboxylase by the modified avidin was partially reversed with an excess of biotin. Attempts to purify the biotin protein in its catalytically active state using modified avidin columns were without success.
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  • 24
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    Archives of microbiology 159 (1993), S. 114-118 
    ISSN: 1432-072X
    Keywords: Bacillus pulvifaciens ; Vegetative cells ; Spotes ; Ultrastructure ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ultrastructure of vegetative cells and spores of Bacillus pulvifaciens was studied by CTEM and SEM methods. The vegetative cells are rods, 1.6–4.5 μm long and 0.4–0.6 μm wide, exhibiting typical ultrastructural features of Gram-positive bacteria. The spores are of ellipsoidal shape, 0.6×1.2 μm in size, with six longitudinal ribs reaching up to 130 nm in height. There are satelite ribs on both sides of the longitudinal ribs, reaching up to 20 nm in height. Between the longitudinal ribs, additional transversal ribs were observed in SEM. A special tubular layer, separating the outer and inner coat of the spores, was revealed in ultrathin sections. This layer seems to be a typical ultrastructural feature of Bacillus pulvifaciens spores.
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  • 25
    ISSN: 1432-072X
    Keywords: Lactobacillus ; Medium composition ; Metal cations ; Electron microscopy ; Protoplast-like forms
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The growth of some locally isolated Lactobacillus strains forming D(-) or L(+) lactic acid, Lactobacillus helveticus ATCC 15009 and Lactobacillus delbrueckii subsp. bulgaricus ATCC 11842 was examined in different media. L. helveticus and Lactobacillus LBL strains formed atypical protoplast-like cells in LAPT medium, sensitive to SDS and proteinase. Specific morphological changes in the cell wall structure of these variants were revealed by transmission and scanning electron microscopy. The effect of glucose and various salts on their appearance was investigated. The prevalent role of metal cations, especially of Mg2+, was established.
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  • 26
    Electronic Resource
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    Archives of microbiology 160 (1993), S. 206-213 
    ISSN: 1432-072X
    Keywords: Treponema denticola ; Spirochetes ; Ultrastructure ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The formation of quasi-multicellular bodies of Treponema denticola was analysed using different electron microscopical methods. These bacteria could develop four different conformations: (i) normal helical forms; (ii) twisted spirochetes, forming plaits; (iii) twisted spirochetes, forming club-like structures; (iv) spherical bodies in different size. Treponemes within spherical bodies, plaits, and clubs proved to be enclosed in a common outer sheath in which the normal arrangement of their axial flagella was lost. The development of the quasi-multicellular bodies starting from the monoforme spirochetes was elucidated and this morphogenetic process is illustrated by a schematic drawing. Factors which might be involved in the induction of the structures are discussed and their possible pathogenetic importance is considered.
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  • 27
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    Cell & tissue research 272 (1993), S. 59-70 
    ISSN: 1432-0878
    Keywords: Sensory cells ; Taste organ ; Electron microscopy ; Bombina orientalis, Rana pipiens (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The taste disc of the red-bellied toad Bombina orientalis (Discoglossidae) has been investigated by light and electron microscopy and compared with that of Rana pipiens (Ranidae). Unlike the frog, B. orientalis possesses a disc-shaped tongue that cannot be ejected for capture of prey. The taste discs are located on the top of fungiform papillae. They are smaller than those in Ranidae, and are not surrounded by a ring of ciliated cells. Ultrastructurally, five types of cells can be identified (mucus cells, wing cells, sensory cells, and both Merkel cell-like basal cells and undifferentiated basal cells). Mucus cells are the main secretory cells of the taste disc and occupy most of the surface area. Their basal processes do not synapse on nerve fibers. Wing cells have sheet-like apical processes and envelop the mucus cells. They contain lysosomes and multivesicular bodies. Two types of sensory cells reach the surface of the taste disc; apically, they are distinguished by either a brush-like arrangement of microvilli or a rod-like protrusion. They are invaginated into lateral folds of mucus cells and wing cells. In contrast to the situation in R. pipiens, sensory cells of B. orientalis do not contain dark secretory granules in the perinuclear region. Synaptic connections occur between sensory cells (presynaptic sites) and nerve fibers. Merkel cell-like basal cells do not synapse onto sensory cells, but synapse-like connections exist between Merkel cell-like basal cells (presynaptic site) and nerve fibers.
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  • 28
    ISSN: 1432-0878
    Keywords: Neurons ; Immunofluorescence ; Tubulin ; Electron microscopy ; Chemoreceptors ; Mechanoreceptors ; Aplysia californica (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The peripheral processes of the mechanoafferents that, when stimulated, initiate the much-studied tail withdrawal reflex of Aplysia californica have not been characterized. We show that immunofluorescence staining for class III β-tubulin highlights neurons and reveals nerve tracts and fine neuronal processes in Aplysia tissue. Coupled with transmission and scanning electron microscopy, class III β-tubulin immunofluorescence is consistent with the possibility that mechanoafferents in the receptive field of pleural ganglion mechanosensory neurons penetrate the tail epidermis and terminate as ciliated endings. This view is reinforced by comparisons among neuronal processes in several mechanosensory epidermal regions and in a chemosensory epidermis.
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  • 29
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    Cell & tissue research 271 (1993), S. 47-57 
    ISSN: 1432-0878
    Keywords: Brush cells ; Cell isolation ; Stomach ; Polarity ; Light microscopy ; Electron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The brush cells (BC) are highly polarized elements occurring in epithelia of endodermal origin. They have a preferential topographical distribution in the organs in which they reside. In the stomach of the rat, BC prevail near the transitional zone separating the forestomach from the glandular stomach. Thus, a method was developed to isolate and recover BC from this organ with the aim of investigating the changes they may undergo after dissociation. Strips of the rat stomach were severed from the very proximal border of the glandular region and incubated in Hanks' balanced salt solution containing pronase. After sedimentation of the dissociated cells (crude sediment containing all stomach epithelial cell types) two successive cell fractions were prepared on preformed Percoll gradient in an attempt to enrich BC in a defined layer. BC were recovered in a fraction at a density close to 1.03 g/ml where they represented about 2% of all cells. The isolated BC changed their form from columnar to pear-shaped; however, they maintained their structural polarity over 2 h as demonstrated by light microscopy, transmission-and scanning-electron microscopy. The fine structure of BC was always satisfactorily preserved. Maintenance of the structural polarity of isolated BC is contrary to the general rule according to which all conventional epithelial cells examined to date lose their polarity after isolation. This result is discussed in relation to morphological findings in isolated sensory cells (hair cells, photoreceptor cells) leading to the suggestion that BC are more similar to these than to conventional epithelial cells.
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  • 30
    ISSN: 1432-0878
    Keywords: Testis ; Electron microscopy ; Cathodoluminescence ; Lipid droplets ; Cholesterol esters ; Vitamin A esters ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Cathodoluminescence (CL) from lipid droplets (LDs) in the rat testis was examined by analytical color fluorescence electron microscopy. The results show that (1) the Cl at wavelengths of 320 nm (CL320) and 450 nm (CL450) is derived from cholesterol esters and a mixture of lipids including vitamin A esters, respectively; (2) CL320 in the LDs of Leydig cells sharply decreases on postnatal day 21, while CL320 and CL450 in the LDs of Sertoli cells begin to be detectable; (3) the CL450-emitting LDs in seminiferous tubules, whose distributional patterns display cyclic changes during the spermatogenic cycle, are involved in spermatogenesis; and (4) the intensity of CL as well as the distributional patterns of CL-emitting LDs in testicular cells change after hypophysectomy, vitamin-A deficiency, and treatment with ethylene dimethane sulfonate and testosterone propionate. This study demonstrates that analytical color fluorescence electron microscopy is a useful tool for in-vivo observation of some specific compounds which cannot be visualized by other methods.
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  • 31
    ISSN: 1432-0878
    Keywords: Pituitary ; Galanin ; Neuromedin-U ; Corticotropes ; Immunocytochemistry ; Electron microscopy ; Plasticity ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization of galanin in rat lactotropes and human corticotropes is well established. Neuromedin U immunoreactivity is present in rat corticotropes but radioimmunoassay of thyroid-manipulated rat pituitaries has also linked it to the thyroid axis. We found galanin immunoreactivity in some rat corticotropes, so we have re-examined rat anterior pituitary galanin- and neuromedin U-like immunoreactivity by use of immunocytochemistry and electron microscopy in rats in the normal state and after estrogen administration or adrenalectomy. In normal rats galanin immunoreactivity was present in a few corticotropes and lactotropes, females showing more than males; neuromedin U-like immunoreactivity was present in some thyrotropes and most corticotropes, in both sexes. Where galanin, neuromedin U and ACTH immunoreactivities were colocalized in corticotropes they were present in the same granules. Estrogen administration caused an increase in number of galanin immunoreactive lactotropes, as previously shown. The proportion of neuromedin U-positive corticotropes was not affected. After adrenalectomy, only females showed a significant increase in the proportion of galanin-positive corticotropes. Neuromedin U immunoreactivity was significantly increased in both sexes, as previously shown. Thus, in rat, as in man, galanin can be present in corticotropes and its expression appears to be sexrelated. This finding, and the demonstration of thyrotrope neuromedin U (only examined in normal females), provide correlation with previous experiments. The influence of endocrine status on the expression of these novel peptides underlines the inherent plasticity of pituitary endocrine cells.
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  • 32
    ISSN: 1432-0878
    Keywords: Aorta ; Endothelium ; Anchoring filaments ; Microfibrils ; Elastin ; Electron microscopy ; Mouse (C57/BL)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The ultrastructural association of endothelial cells with the subjacent elastic lamina was investigated in the developing mouse aorta by electron microscopy. In the 5-day postnatal aorta, extensive filament bundles extend along the subendothelial matrix connecting the endothelial cells to the underlying elastic lamina. The connecting filaments form lateral associations with the abluminal surface of the endothelial cells in regions of membrane occupied by membrane-associated dense plaques. On the intracellular face of each plaque, the termini of stress fibers penetrate and anchor to the cell membrane in alignment with the extracellular connecting filaments. Both the stress fibers and the connecting filaments are oriented parallel to the longitudinal axis of the vessel. High magnification electron micrographs of individual endothelial cell connecting filaments reveal features similar to those of elastin-associated microfibrils. Each connecting filament consists of a 9–10 nm linear core with an electron-lucent center and peripheral spike-like projections. From the filaments, small thread-like extensions span laterally, linking the filaments into a loose bundle and anchoring them to the endothelial cell membrane and the surface of the elastic lamina. The filaments also appear heavily coated with electron-dense material; often with some degree of periodicity along the filament length. During development, the number of endothelial cell connecting filaments decreases as the elastic lamina expands and the subendothelial matrix is reduced. In the aortic intima of mature mice, the elastic lamina is closely apposed to the abluminal surface of the endothelial cell and no connecting filaments are seen. These observations suggest that endothelial cell connecting filaments are developmental features of the aortic intima which, together with the intracellular stress fibers, aid to maintain the structural integrity of the endothelial cell layer during development by providing the cells with protection from intraluminal shear forces.
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  • 33
    ISSN: 1432-0878
    Keywords: Corpora allata ; Electron microscopy ; Morphometry ; Ovariectomy ; Juvenile hormone ; Cockroach, Diploptera punctata (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Morphometric studies were made on corpora allata of the cockroach Diploptera punctata from animals in which increasing gland size is not coupled to hormone synthesis (ovariectomized mated females; last-instar larvae) and in which gland size is coupled to hormone synthesis (normal mated and virgin females; penultimate-instar larvae). Cell number, gland volume, and juvenile hormone synthesis were measured. From electron micrographs, nuclear, cytoplasmic, and extracellular volumes; and cell membrane area were calculated; and fine structure described. Low-activity glands of ovariectomized mated females resembled high-activity glands from mated females in high cell number, large overall and cytoplasmic volume, and low nuclear-cytoplasmic ratio; they differed in having organelles typical of low-activity glands, mitochondria with dense matrices and large whorls of smooth endoplasmic reticulum. Inactive lastinstar larval glands resembled mated ovariectomized, female glands in increased cell number and organelles characteristic of inactive glands; however, their nuclearcytoplasmic volume ratio was much higher. Penultimate cytoplasmic volume ratio was much higher. Penultimate larval glands with high activity per cell resembled active glands of normal mated females. Ovariectomy did not change morphometric parameters of virgin female glands; thus mating results in increase in size of adult female glands whereas the growing ovary is needed for changes in mitochondria and endoplasmic reticulum associated with high juvenile hormone synthesis.
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  • 34
    ISSN: 1432-0878
    Keywords: Peripolar cells ; Juxtaglomerular apparatus ; Cytoplasmic granules ; Exocytosis ; Electron microscopy ; Sheep, newborn
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Scanning and transmission electron microscopy were used to study the ultrastructural characteristics and positions of granulated peripolar cells in newborn lamb kidney. Following tissue fixation by vascular perfusion in situ, the vascular pole region of the glomerulus was exposed for examination by scanning electron micoscopy following removal of the glomerular tuft. Peripolar cells were recognized by their surface morphology enabling their quantification and an assessment of the relationship of their position in the renal cortex. The prominent expression of peripolar cells in this species was confirmed. Almost every vascular pole examined revealed peripolar cells (405 out of 407; 99.5%) and thus, throughout the cortex, the distribution of peripolar cells was the same as the distribution of renal corpuscles. Larger, more protruding peripolar cells were observed in the outer cortical renal corpuscles. The numbers of peripolar cells encircling each vascular pole ranged from 1 to 10. There was no correlation between number of granulated peripolar cells at the vascular pole and the position of the renal corpuscle within the renal cortex. As viewed by transmission electron microscopy, organelles of protein synthesis were abundant in the cytoplasm of peripolar cells. Exocytosis of cytoplasmic granules was observed by both scanning and transmission electron microscopy implying that a process of regulative secretion occurs from these cells. The use of ultrastrural techniques has provided evidence supporting the concept that peripolar cells are prominent in the cuff region of each renal corpuscle of the newborn lamb and further-more that peripolar cells in this species most likely have a secretory function.
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  • 35
    ISSN: 1432-0878
    Keywords: Collagen fibril ; Three dimensional cell culture ; Ascorbate ; Aminopropeptide, type I ; Aminopropeptide, type III ; Electron microscopy ; Immunoelectron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The purpose of this study was to follow collagen fibril formation in a newly developed three dimensional cell culture system. Human neonatal foreskin fibroblasts were grown on a nylon mesh in Dulbecco's Modified Eagles Medium (DMEM) supplemented with 10% fetal calf serum and antibiotics. Fibrillogenesis was initiated by the addition of 50 micrograms/ml ascorbate to confluent cultures. Sample meshes were processed for electron microscopy or immuno-electron microscopy. Fibrils ≈20–30 nm in diameter, with 67 nm periodicity, were first detected five days after the addition of ascorbate. As cultures progressed, cells organized into parallel layers between which collagen fibers continued to form and increase in diameter. By day 50, fiber diameter ranged from 30 to 80 nm and large bundles were seen. No collagen fibril formation occurred in control cultures to which no ascorbate was added. However, large amounts of microfibrils were observed. Antibodies against the aminopropeptide of type I procollagen were found to bind to fibrils with diameters less than 34 nm while antibodies against the aminopropeptide of type III collagen bound primarily to fibers which ranged from 35–54 nm in diameter. We believe that this system, which morphologically resembles a normal dermis, will werve as an excellent model for the study of collagen fibrillogenesis.
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  • 36
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    Cell & tissue research 272 (1993), S. 447-457 
    ISSN: 1432-0878
    Keywords: Horizontal cells ; Calcium-binding protein ; Synaptie input ; Inner plexiform layer ; Immunoreactivity ; Electron microscopy ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Bovine retinae were stained immunocytochemically with antibodies against the calcium-binding protein, calbindin. Horizontal cells in the outer plexiform layer were heavily labelled. The processes of most horizontal cells were confined to the level of the outer plexiform layer, and the tips of their dendrites were positioned as the lateral elements of the cone triads, viz. the usual mammalian arrangement. However, some of the horizontal cells had additional thick processes descending to branch within the inner plexiform layer, where they were postsynaptic at bipolar cell dyads and where they also received input from amacrine cells. No output synapses of horizontal cells were observed in the inner plexiform layer.
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  • 37
    ISSN: 1432-0878
    Keywords: Basic fibroblast growth factor ; Trigeminal ganglion ; Immunohistochemistry ; Electron microscopy ; In situ hybridization histochemistry ; Vibrissae ; Hair ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have characterized an antiserum against basic fibroblast growth factor (bFGF) by immunoblot, investigated the location of bFGF-like immunoreactivity (bFGF-IR) in the trigeminal sensory system and perioral skin endowed with vibrissae, and demonstrated the site of bFGF mRNA expression in the vibrissae by in situ hybridization histochemistry. Light-microscopic immunohistochemistry has demonstrated that bFGF-IR is present not only in trigeminal ganglion neurons and their central and peripheral processes, but also in cells of the matrix, external root sheath and papillae of vibrissae and the stratum basale of the stratified squamous epithelium of the skin. Electron microscopy has revealed intense bFGF-IR mainly in cytoplasmic regions, other than the lumen of rough endoplasmic reticulum and the Golgi apparatus, in trigeminal ganglion neurons, in fibroblast-like cells in the papillae, and in capsules of vibrissae. In contrast, actively proliferating and/or differentiating cells in the matrix of vibrissae have intensely stained euchromatin and weakly labeled cytoplasm that, unlike that of the aforementioned cells, contain immunoreaction products in discrete spots less than 100 nm in diameter, implying the generation of different molecular forms of bFGF in cells of the matrix and papillae. Moreover, the accumulation of bFGF in the euchromatin appears to take place in cells at non-mitotic stages (possibly interphases), characterized by a conspicuous nucleolus and well-developed nuclear envelope. A digoxigenin-labeled cRNA probe for the demonstration of bFGF mRNA gives conspicuous hybridization signals mainly in the matrix of vibrissae. These findings suggest that bFGF is involved in the growth and differentiation of matrix cells during certain periods of the cell cycle and that it acts as a non-mitogenic mediator in the adult trigeminal sensory system.
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  • 38
    ISSN: 1432-0878
    Keywords: Adrenal growth ; CRH ; ACTH ; Hypophysectomy ; Electron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Within two weeks, hypophysectomy induced in rats a striking decrease in the level of circulating ACTH (the concentration of which was at the limit of sensitivity of our assay system), coupled with a net reduction in the plasma corticosterone concentration and an evident adrenal atrophy. Zona fasciculata, the main producer of glucocorticoids, was decreased in volume, due to a lowering in both the number and average volume of its parenchymal cells. Subcutaneous ACTH infusion (0.1 pmol·min-1), administered during the last week following hypophysectomy, restored the normal blood level of ACTH and completely reversed all effects of hypophysectomy on the adrenals. Subcutaneous infusion for one week with α-helical-CRH or corticotropin-inhibiting peptide (1 nmol·min-1), which are competitive inhibitors of CRH and ACTH, evoked a further significant lowering of plasma corticosterone concentration and markedly enhanced adrenal atrophy in hypophysectomized rats. These findings strongly suggest that an extrahypothalamic pituitary CRH/ACTH system may be involved in the maintenance of the growth and steroidogenic secretory activity of the rat adrenal cortex.
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  • 39
    ISSN: 1432-0878
    Keywords: Serotonin ; Alimentary canal, insect ; Stomatogastric neryous system ; Immunohistochemistry ; Bioassay ; Electron microscopy ; Leptinotarsa decemlineata (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical studies showed that the alimentary canal of Leptinotarsa decemlineata receives serotoninergic innervation from different neurons in the central and stomatogastric nervous system. The foregut is innervated by the frontal ganglion. Four of the 6–8 large neurons present in this ganglion have axons which run to the musculature of the oesophagus, crop, sphincter, and frontal area of the midgut. They are accompanied by axons from neurons in the suboesophageal ganglion, and by axons from as yet unidentified non-immunoreactive neurons in thebrain and/or the ventral nerve cord. The posterior part of the midgut is essentially devoid of serotoninergic innervation. The hindgut is innervated by two large neurons in the caudal tip of the last abdominal ganglion. The axons always run to the circular and longitudinal muscles of the crop, the circular muscles of the sphincter, and the longitudinal muscles of the hindgut. Immunohistochemical electron microscopy suggests that exocytosis of the immuno-labelled vesicles may occur at some distance from the muscle fibres, implying a neurohormonal release of this neurochemical. A bioassay used to demonstrate the type of effect of serotonin on isolated hindguts in vitro, indicated a clear inhibitory effect on spontaneous contractions at concentrations of 10-8–10-5 M. This effect was dose-dependent. Axons found in association with the cryptonephridial system on the hindgut might be involved in the control of diuresis although we have not tested this possibility experimentally.
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  • 40
    ISSN: 1432-0878
    Keywords: Serotonin (5-HT) ; Neurohemal systems ; Immunohistochemistry ; Electron microscopy ; Targeted release ; Leptinotarsa decemlineata (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We analyzed the anatomy of two diffuse neurohemal systems for serotonin in the head of the Colorado potato beetle Leptinotarsa decemlineata by means of immunohistochemistry. One system is formed by axons from two bilateral pairs of neurons in the frontal margin of the suboesophageal ganglion that enter the ipsilateral mandibular nerve, emerge from this nerve at some distance from the suboesophageal ganglion, and cover all branches of the mandibular nerve with a dense plexus of immunoreactive axon swellings. The other system is formed by axons from two large neurons in the frontal ganglion that enter the ipsilateral frontal connectives, emerge from these connectives, and form a network of axon swellings on the labroforntal, pharyngeal, and antennal nerves and on the surface of the frontal ganglion. Immunohistochemical electron microscopy demonstrated that the axon swellings are located outside the neural sheaths of the nerves and hence in close contact with the hemolymph. We therefore suggest that these plexuses represent extensive neurohemal systems for serotonin. Most immunoreactive terminals are in direct contact with the hemolymph, and other terminals are closely associated with the muscles of the mandibles, labrum, and anterior pharynx, as well as with the salivary glands, indicating that these organs are under serotoninergic control.
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  • 41
    ISSN: 1615-6102
    Keywords: Barley yellow mosaic virus ; Cytoplasmic inclusion bodies ; Electron microscopy ; Hordeum vulgare ; Immunogold labeling ; RNA 2-encoded proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Antisera were raised against the RNA 2-encoded proteins of 28 kDa and 70 kDa of barley yellow mosaic virus (BaYMV) by using the corresponding cDNA sequences of a German isolate for protein overexpression inEscherichia coli BL 21 and subsequent purification. The proposed processing of a 98 kDa precursor polyprotein encoded by the long open reading frame of RNA 2 to two proteins of 28 kDa and 70 kDa could be confirmed by immunoprecipitation of the in vitro transcribed and translated cDNA-clone of RNA 2 and Western blot analysis of fragmentated protein extracts of BaYMV-infected winter barley plants. In situ localisation studies of infected leaf tissue using immunogold labeling techniques for electron microscopy revealed that both viral proteins of BaYMV (RNA 2) were associated with the crystal-like cytoplasmic inclusion bodies. No other parts of the cells and no other inclusions (pinwheelstructures or aggregated virus particles) showed any gold labeling when the 28 kDa and 70 kDa antisera were used. We suppose that both RNA 2-encoded proteins take part in the formation of the crystal-like cytoplasmic inclusion bodies which are the most dominant structures in the cytoplasm of BaYMV-infected tissue. Possible functions of the 28 kDa and 70 kDa protein of BaYMV (RNA 2) are discussed.
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  • 42
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    World journal of microbiology and biotechnology 9 (1993), S. 108-112 
    ISSN: 1573-0972
    Keywords: Electron microscopy ; celluloytic microorganisms ; termite gut
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The major gut microflora colonizing the hind gut of a higher termite,Odontotermes obesus, included morphologically diverse bacteria, both coccoid and rod-shaped, along with spirochaetes, pseudomonads and actinomycetes. Flagellated protozoa were totally absent. When the gut extract was inoculated on plates containing carboxymethyl cellulose or cellobiose, higher numbers of bacteria grew than on plates without cellulosic sources. The gut homogenate exhibited strong hydrolytic activity when carboxymethyl cellulose,p-nitrophenyl-β-d-glucoside or xylan were used as substrate, indicating the role of gut microbiota in the process of cellulose and hemicellulose digestion. Activities were highest in the hind gut, and the paunch was probably the major site of polysaccharide digestion in this higher termite.In vitro cultivation of some of the isolates revealed both cellulase and xylanase activities. To our knowledge, this is the first report on ultrastructural studies of the higher termiteOdontotermes obesus.
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  • 43
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    Documenta ophthalmologica 84 (1993), S. 351-363 
    ISSN: 1573-2622
    Keywords: Cell culture ; Electron microscopy ; Human lens epithelium ; Morphology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A system for culturing human lens epithelial cells in the laboratory was developed. The morphological appearances of the cells was studied using phase contrast, scanning and transmission electron microscopy. Cell marker studies using monoclonal antibodies to cyto-keratin, vimentin and epithelial membrane antigen were also performed. There was a marked increase in cell size as a function of time in culture. After 3 to 4 weeks cells showed early signs of ageing. By 6 to 8 weeks the majority of the cells had become very irregular in shape and demonstrated irregularities of the plasma membrane and intra-cytoplasmic vacuole formation. The cells stained strongly for vimentin and epithelial membrane antigen. Staining with cytokeratin was somewhat weaker. This culture technique provides us with a suitable model for studying the growth behavior of these cells.
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  • 44
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    European archives of oto-rhino-laryngology and head & neck 249 (1993), S. 499-502 
    ISSN: 1434-4726
    Keywords: Hypopharynx-Synovial sarcoma ; Supraglottic laryngopharyngectomy ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Synovial sarcoma in an extra-articular location is a very rare tumor, particularly in relation to childhood tumors. We report our experiences with an 11-year-old boy who suffered from swallowing difficulties caused by a polypoid tumor of the hypopharynx. Biopsy demonstrated a fusocellular carcinoma, while the epithelial components showed cytokeratin and neuron-specific enolase positivity. A correct final diagnosis could only be established by the histology of the surgical preparation following partial laryngopharyngectomy. The presence of spindle cells associated with glandular-like ones proved the synovial character of the tumor. The spindle cells were negative for epithelial marker but were positive for vimentin. S-100 protein positivity could only be demonstrated in the nerve elements encapsulated in the tumor. Ultrastructural examinations confirmed the presence of the different cell types. The spindle cells were rich in intermediate fibers, as demonstrated by electron microscopy.
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  • 45
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    European archives of oto-rhino-laryngology and head & neck 250 (1993), S. 44-50 
    ISSN: 1434-4726
    Keywords: Cochlear glomeruli ; Guinea pig ; Corrosion casting ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The cochlear glomeruli were studied in guinea pigs using scanning electron microscopy of vascular corrosion casts and transmission electron microscopy of tissue sections. Two types of coiled vessels forming the cochlear glomeruli were found in the bony wall of the modiolus. First, upper glomeruli were seen situated in the bony wall next to the scala vestibuli; second, lower glomeruli were located in the osseous spiral lamina just above the spiral ganglion. Upper glomeruli gave rise to radiating arterioles which supplied capillaries of the stria vascularis, while lower glomeruli fed the capillaries of the spiral lamina and limbus. Unlike the main supplying arteries, smooth muscle cells were not present in the walls of the arterioles forming the glomeruli and a peculiar layer of lamellar pericytes was found. The arterioles were strikingly longer than their parent vessels and no autonomic nerves were found in close spatial relationship. Hence, these findings indicate that the cochlear glomeruli serve as efficient devices for reducing cochlear blood pressure.
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  • 46
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    European archives of oto-rhino-laryngology and head & neck 250 (1993), S. 401-407 
    ISSN: 1434-4726
    Keywords: Basement membrane ; Nasal concha ; Nasal mucosa ; Electron microscopy ; Immunofluorescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Light and electron microscopic as well as immunohistochemical investigations were performed on the basement membrane (BM) of the nasal mucosa from the inferior nasal conchae of 20 patients, aged 15–50 years. Two of the patients served as controls. Clinical disorders requiring the turbinectomies were hyperplastic nasal inferior conchae (17 patients) and immobile cilia syndrome (1 patient). In all cases light microscopy demonstrated a 10–15 μm thick homogeneous BM underneath an intact epithelium. Electron microscopic findings revealed a typical subepithelial basal lamina (BL). The remaining parts of the BM consisted of single, isolated 25-mm-thick collagenous fibrils. Only a few cells and small unmyelinated nerves occurred in this layer. Immunohistochemical investigations showed BL components (collagen type IV, laminin, nidogen and heparan sulfate proteoglycan) directly underneath the epithelium. Collagen types I, III, V, and VI could also be demonstrated immunohistochemically in the remaining parts of the BM. Collagen type VII was allocated to the anchor filaments beneath the BL. This special BM is presumed to contribute to mechanical stabilization of the epithelium. The significance of the BL and the subepithelial connective tissue for the behavior of the epithelium — e.g. direction of differentiation - are discussed.
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  • 47
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    European archives of oto-rhino-laryngology and head & neck 250 (1993), S. 412-417 
    ISSN: 1434-4726
    Keywords: Sugar-binding site ; Guinea pig ; Middle ear ; Lipopolysaccharide ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Glucosamine-binding sites were detected in Lowicryl K4M-embedded guinea pig middle ear mucosa by electron microscopy, using glucosaminyl bovine serum albumin. Incubation of ultrathin tissue sections with gold-labeled glucosaminyl bovine serum albumin (GlcN/BSA/gold) resulted in binding mainly on cilia, microvilli, rough endoplasmic reticulum and nuclei. The sugar binding was not inhibited after ultrathin sections had been digested with trypsin or neuraminidase. Various carbohydrates and glycoconjugates were tested as competitive inhibitors of G1cN/BSA/gold labeling on the tissue sections. The sugar specificity range detected by the glucosamine-binding sites included glucosamine, N-acetylglucosamine, mannose and fucose, whereas N-acetylgalactosamine, galactose and glucose were not detectable. A series of endotoxic substances such as Salmonella minnesota Re595 lipid A complex with BSA and lipopolysaccharides (LPS) derived from Escherichia coli 055: B5 or S. minnesota Re595 also competed with GlcN/BSA/gold binding. This indicates that the lipid A backbone glucosamine or other carbohydrate portions of LPS is a part of the structure recognized by glucosamine binding sites.
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  • 48
    ISSN: 1860-1499
    Keywords: Electron microscopy ; Morphometry ; Chronic type lymphoid leukemias
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Malignant cells from 1 case of chronic lymphocytic leukemia of the T-cell type (T-CLL), 5 cases of chronic lymphocytic leukemia of the B-cell type (B-CLL), 1 case of prolymphocytic leukemia of the B-cell type (B-PLL), 1 case of lymphocytic lymphoma of intermediate differentiation of the B-cell type (B-ILL), 1 case of splenic B-cell lymphoma with circulating villous lymphocytes (SLVL), and 2 cases of hairy cell leukemia (HCL) were measured on electron micrographs. The area of the cytoplasm, nucleus, nucleoli, and the perimeters of nuclei were measured and the nucleo-cytoplasmic ratio, nucleolo-nuclear ratio, and nuclear contour index (NCI) were calculated. Malignant cells from B-PLL had the largest cytoplasmic, nuclear, and nucleolar areas. The NCI was largest (4.74) in T-CLL. B-ILL, B-PLL, and SLVL had an intermediate (4.5) NCI. HCL and B-CLL had the smallest (4.1) NCI. Lymphoid cells in SLVL were smaller in size than HCL cells and had a higher NCI than that of HCL. Thus, electron microscopic morphometry can provide further information to aid in distinguishing chronic type lymphoid leukemias.
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  • 49
    ISSN: 1860-1499
    Keywords: Murine model ; Glomerulosclerosis ; GBM alteration ; Proteinuria ; Dense deposits ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Renal pathology of the FGS/Nga strain of mice, which has been known to develop proteinuria and progressive glomerulosclerosis, was studied by light and electron microscopy. Sclerotic changes had increased by 6 months and most of the glomeruli were totally obsolescent by 10 months. By electron microscopy, electron dense deposits were seen in the mice by 3 months and thereafter. Splitting or the irregular thickening of the glomerular basement membrane (GBM) was observed, and some mice showed reticulation in the GBM. The fixed negative charges detectable with polyethyleneimine (PEI) were decreased in the mice with massive proteinuria.
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  • 50
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    Medical molecular morphology 26 (1993), S. 57-63 
    ISSN: 1860-1499
    Keywords: Acute leukemia ; Megakaryoblastic leukemia ; Glycogen ; Glycoconjugates ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The localization of complex carbohydrates in 41 patients with acute megakaryoblastic leukemia (AMKL) was studied using the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method. Examination by the PA-TCH-SP method revealed that the megakaryoblasts from AMKL had a thick cell coat, a small amount of glycogen, granules and well-developed GERL ofNovikoff. Multivariate analysis revealed no cytochemical differences in the distribution of glycoconjugates in the cytoplasm between AMKL blasts in children and adults, but they were noted between all AMKL blasts and blasts from patients with chronic myelocytic leukemia in megakaryoblastic crisis (CML-MK). The ultrastructural distribution of complex carbohydrates in the blasts from AMKL was extremely different from the acute myelocytic leukemia (AML) blasts as demonstrated by the analysis.
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  • 51
    ISSN: 1860-1499
    Keywords: Subarachnoid hemorrhage ; Calcium ; Diltiazem ; Vasospasm ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Calcium channel blockers such as diltiazem are being explored as agents to reduce or prevent cerebral vasospasm following subarachnoid hemorrhage (SAH). Yet little is known concerning changes over time in the calcium ions in the arterial wall after subarachnoid hemorrhage even when calcium channel blockers are used. The results imply that the massive increase of calcium ions in the smooth muscle cells in the early stages of SAH may be related to morphological changes in the spastic vessels in the subacute stage of SAH and calcium ion blockers might be useful in reducing organic changes in the vessels by the inhibition of calcium overloading.
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  • 52
    ISSN: 1860-1499
    Keywords: Mg-ATPase ; Enzyme cytochemistry ; Electron microscopy ; Aspirin ; Clofibrate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A total of 36 male F344 rats was fed a diet containing 1% aspirin or 0.25% clofibrate for 1, 3 and 4 weeks. Hepatomegaly was more evident with aspirin than with clofibrate. The activity of magnesium dependent adenosine triphosphatase (Mg-ATPase) was localized on the limiting membrane of aspirin-induced peroxisomes, but it was much weaker than that in control and clofibrate-induced peroxisomes. Alteration in the ultrastructure of peroxisomes and mitochondria became more evident in Week 4 of the experiment than in Week 1. These ultrastructural changes also basically returned to normal conditions after one week withdrawal of the chemicals.
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  • 53
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    Medical molecular morphology 26 (1993), S. 185-190 
    ISSN: 1860-1499
    Keywords: Medial necrosis ; Cerebral artery ; Hypertension ; Vasospasm ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Medial necrosis is a preceding lesion to angionecrosis which may cause hemorrhage and lacunar infarct. We morphometrically studied the loss of smooth muscle cells in the media of human cerebral arteries and examined the occurrences of vasospasm. Medial necrosis of hypertensive cases was significantly more severe than that of non-hypertensive cases (p〈0.001). The outer media was significantly more vulnerable than the inner one (p〈0.001). Medial necrosis in the perforating artery was slightly more severe than that of the cortical branches. Vasospasms were identified electron microscopically in 4 of 18 cases. These findings may satisfactorily explain the possibility of vasospasm as a cause of medial necrosis.
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  • 54
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    Primates 34 (1993), S. 233-235 
    ISSN: 0032-8332
    Keywords: Chimpanzee ; Infant-eating ; Electron microscopy ; Feces ; Hair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A large bolus of hairs found in the feces of an adult wild chimpanzee in the Budongo Forest, Uganda, was identified as belonging to a chimpanzee below the age of 3 yrs. This represents the second case of infant-eating recorded in the Budongo Forest.
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  • 55
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    The @Anatomical Record 235 (1993), S. 533-538 
    ISSN: 0003-276X
    Keywords: Permeability barrier ; Tight junction ; Fusiform vesicle ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Formation of the blood-urine permeability barrier in differentiating mouse transitional urothelium was studied. It was established that the development of superficial cell barrier is a two-phase process: beginning with formation of the tight junctions, followed by formation of fusiform vesicles and asymmetric apical plasma membranes. Fusiform vesicles differentiate during days 15 and 17 of gestation and fuse with the apical plasmalemma. Thus a thick membrane is formed before the excretion of hypertonic urine into the embryonic bladder. Through some degenerative superficial cells slough between fetal day 17 and the day of birth, the bladder epithelium in mice does not lack an effective permeability barrier. © 1993 Wiley-Liss, Inc.
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  • 56
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    The @Anatomical Record 236 (1993), S. 593-601 
    ISSN: 0003-276X
    Keywords: Main excretory duct epithelium ; Lateral interdigitations ; Goblet cells ; Fenestrated capillaries ; Pored domes ; Submandibular gland ; Japanese monkey ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The fine structure and subepithelial capillaries of the main excretory duct epithelium (MEDE) of the submandibular gland in the Japanese monkey, Macaca fuscata were investigated by light, scanning, and transmission electron microscopy. Capillaries found beneath the MEDE were all fenestrated ones. Some of them had elevated endothelium known as pored domes. The capillary density (number of capillaries per 200 μm length of MEDE) was 6.05 ± 1.36, and the number of fenestrae per 10 μm length of available endothelium was 11.62 ± 3.82. The MEDE was pseudostratified, and consisted of four cell types: Type I and II, which were columnar cells without basal infoldings, basal and goblet cells. The most characteristic feature was the presence of well-developed lateral interdigitations between Type I and II cells. © 1993 Wiley-Liss, Inc.
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  • 57
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    The @Anatomical Record 235 (1993), S. 253-260 
    ISSN: 0003-276X
    Keywords: Electron microscopy ; Endothelial cell ; Endothelin ; Sarafotoxin ; Toad aorta ; Vasocontraction ; Weibel-Palade body ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Dose-dependent tension curves were recorded from the in vitro toad aortas by administration of endothelin-1 and sarafotoxin-S6b. The maximal contractile tensions by both drugs were evoked at a 10-8 M concentration. By a single dose application (10-8 M) of endothelin-1 and sarafotoxin-S6b to both endothelium-preserved and denuded vessels, the induction of the endothelium-dependent vasocontraction occurs after 2 min of administration.Ultrastructural changes of Weibel-Palade bodies such as decrease in electron density, swelling with a wide peripheral halo, and expulsion of their contents in a manner of exocytosis become evident within 2 min after administration of these drugs. These findings indicate that some vasocontractile substances in Weibel-Palade bodies are extracellularly discharged by endothelin-1 and sarafotoxin-S6b. © 1993 Wiley-Liss, Inc.
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  • 58
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    The @Anatomical Record 236 (1993), S. 70-86 
    ISSN: 0003-276X
    Keywords: Comparative histology ; Light microscopy ; Electron microscopy ; Cytochemistry ; Immunocytochemistry ; Neuroepithelial endocrine cells ; Neuroepithelial bodies ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 22 Ill.
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  • 59
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    The @Anatomical Record 236 (1993), S. 721-729 
    ISSN: 0003-276X
    Keywords: Macaca mulatta; Area 17 ; Neurons ; Age changes ; Electron microscopy ; Light microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: We have assessed the effects of aging on the cell bodies of the large Meynert cells in layers V and VI of the rhesus monkey primary visual cortex by comparing the frequency, size, and cytology of these neurons in three young (5-6 years of age), one middle-aged (12 years of age), and four old monkeys (25-35 years of age). In the young monkeys there is an average of 20.8 Meynert cells beneath 1 mm2 of cortical surface, and in the old monkeys the value is 22.3 cells. The cell bodies of these large pyramidal cells do not become smaller with age, and surprisingly they accumulate only small amounts of lipofuscin. The conclusion is that in the rhesus monkey there is no loss of Meynert cells with age and that aging has little effect on the morphology of the cell bodies of these neurons. © 1993 Wiley-Liss, Inc.
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  • 60
    ISSN: 0003-276X
    Keywords: Urethral glands ; Testosterone ; Morphometry ; Electron microscopy ; Male mice ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Recent studies of the urethral glands in the male mouse and rat have suggested that they are testosterone-dependent glands that may be potential sites for secretory immunity in the male genital tract. In the present study we describe the ultrastructural features of these glands in normal mice and provide quantitative data on the sizes of the acinar cells and their organelles in sham-, oil-, and testosterone-treated castrated mice. Acinar cells in urethral glands from normal mice contain numerous secretory granules, prominent Golgi complexes, elongated mitochondria, and an abundance of rough endoplasmic reticulum (RER) with large and dilated cisternae, all of which are features characteristic of secretory cells. In some acinar cells the cisternae of the RER were filled with closely packed, unbranched, straight, tubular structures that were oriented parallel to one another, that radiated from aggregates of dense material, or that were randomly attanged. In other acinar cells the cisternae of the RER showed a network of branching and anastomosing vesicular-like structures whose limiting membranes were occasionally seen in continuity with the membranes of the RER. Secretory acini showed large, unbranched tubules in the acinar lumen. When cut at right angles the large tubules exhibited a distinct fuzzy outer coat with fine projections radiating outwards. The ultrastructure of the acinar cells and the presence of tubules in the lumen suggests that they are engaged in secretion of a tubular protein.Morphometric analysis of acinar cells in the urethral glands showed that the mean volumes of nuclei, cytoplasm, secretory granules, vacuoles, and mitochondria were significantly reduced in castrated mice in comparison to either normal or testosterone-treated castrated mice. This confirms earlier observations that the urethral glands are targets of testosterone. © 1993 Wiley-Liss, Inc.
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  • 61
    ISSN: 1059-910X
    Keywords: In situ hybridization ; Digoxigenin ; Electron microscopy ; Cryosections ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The technique of in situ hybridization has been used to evaluate the expression of an ovulation hormone mRNA (caudodorsal cell hormone; CDCH) in the central nervous system (CNS) of the mollusc Lymnaea stagnalis. Hybridization with radioactive as well as with nonradioactive labeled oligonucleotide and plasmid probes revealed a specific labeling on cell bodies of caudodorsal cells (CDCs), which are known to produce CDCH, on the light microscopical level. In addition, specific labeling was observed outside the cell bodies, as far as the cerebral commissure, where CDCH is released in the haemolymph. To investigate whether these signals represent an axonal localization of the CDCH mRNA, we performed in situ hybridization at the electron microscopical (EM) level. The results showed an intraaxonal localization of CDCH mRNA with digoxigenin labeled oligonucleotide and plasmid probes. Gold labeling was observed in secretion granules, and double labeling experiments showed that these granules also contain CDCH. This specific intragranular localization suggest that CDCH mRNA is transported through the axon and released by exocytosis in the haemolymph. © 1993 Wiley-Liss, Inc.
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  • 62
    ISSN: 1059-910X
    Keywords: Double-stranded viral DNA ; Electron microscopy ; HeLa cell ; In situ hybridization ; Lytic infection ; S1 nuclease ; Replication ; Viral Ad5 genomes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In order to gain a further insight into the relationships of the complex process of replication of adenovirus genomes to the substructures which occur in the nuclei of adenovirus type 5 (Ad5) infected HeLa cells, we have visualized directly, at the electron microscopic level, viral double-stranded DNA (dsDNA) in late infected nuclei by the use of a post-embedding in situ hybridization technique with a biotinylated specific DNA probe. The procedure is based on the removal of single-stranded (ss) nucleic acids by S1 nuclease. The highest levels of signal density for viral dsDNA were detected over the fibrils of the large, centrally located viral genome storage site and over the viral nucleoids of both clustered and isolated viruses. Lower but significant signals were observed over the fibrillo-granular network of the peripheral replicative zones, where both transcription and replication of viral DNA occur. On the other hand, the labeling of the enclosed viral ssDNA accumulation sites, also involved in viral replication but not transcription, was negligible, which suggests that, in the latter, the newly synthesized viral dsDNA immediately extends into the adjacent peripheral replicative zone to be transcribed and/or replicated.
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  • 63
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    Microscopy Research and Technique 25 (1993), S. 429-433 
    ISSN: 1059-910X
    Keywords: Biocorrosion ; Sulfate-reducing bacteria ; Biofilm ; Desulfovibrio ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The biofilm attributed to Desulfovibrio vulgaris growing in the presence of ferrous metals was examined with an environmental scanning electron microscope. This novel microscope produced images of iron sulfide colloids and other iron containing structures that had not been reported previously. A plaque composed of iron sulfide enveloped the surface of the corroding metal while crystals containing magnesium, iron, sulfur, and phosphorus were present in the culture where corrosion was in progress. A structure resembling the tubercule found in aerobic corrosion was observed on stainless steel undergoing biocorrosion and the elements present in this structure included sulfur, iron, chloride, calcium, potassium, and chromium. © 1993 Wiley-Liss, Inc.
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  • 64
    ISSN: 1059-910X
    Keywords: Microwave fixation ; Freeze-fracture ; Electron microscopy ; Protozoan ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Tritrichomonas foetus, a pathogenic protozoan, was used as a model to analyse microwave-stimulated fixation as a procedure of preparation of biological samples for electron microscopy of thin sections and freeze-fracture replicas. Good preservation of the protozoan structure was achieved by microwave-stimulated fixation and Epon polymerization. The membrane structure, as visualized in freeze-fracture replicas, was well preserved. © 1993 Wiley-Liss, Inc.
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  • 65
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    Microscopy Research and Technique 25 (1993), S. 46-60 
    ISSN: 1059-910X
    Keywords: Vasopressin ; Vasoactive intestinal polypeptide ; Gastrin releasing peptide ; GABA ; In vitro ; In situ hybridization histochemistry ; Electron microscopy ; Circadian pacemaker ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Suprachiasmatic nuclei (SCN) from hypothalami of postnatal rats were maintained for 18-39 days in vitro as organotypic slice explants. Neuronal subtypes containing vasopressin (VP), vasoactive intestinal polypeptide (VIP), gastrin releasing hormone (GRP), and GABA were immunocytochemically identifiable in these cultures. In situ hybridization histochemistry was compatible with these SCN slice explant cultures, and mRNA encoding for VP was detected bilaterally within these nuclei. After 18 days in vitro, both VP mRNA and VP immunoreactivity increased from levels present on postnatal days 4 (the earliest age from which the explanted tissue was derived) to levels typical of adult SCNs. In contrast, the GRP expression remained low, characteristic of early postnatal animals and far lower than adult levels. This suggests that the developmental cues or programs necessary for enhanced VP expression are maintained in these cultures, while those affecting GRP expression are absent or inhibited. VIP-containing neurons were numerous in the cultures. Culture slices appeared healthy, and similar numbers and distributions of identifiable neurons within the SCN were observed, whether or not the slices were grown in the presence of serum. EM analysis revealed that the SCN in vitro is composed of tightly packed neurons, processes, and abundant synapses containing both clear and dense core vesicles, closely resembling the SCN in vivo. Vasopressinergic neuronal somata contained extensive Golgi systems and labeled secretory granules, the latter organelle being present also within processes and synaptic terminals. GABA-immunopositive processes and synaptic profiles were abundant, with labeling occurring particularly over secretory vesicles and mitochondria. This slice culture system effectively maintained much of the intrinsic organization and cellular components of the SCN for long periods in vitro and should be an excellent model system for studying the intrinsic molecular mechanisms and extrinsic cues which regulate neuronal phenotype in this circadian pacemaker. Published 1993 Wiley-Liss, Inc.
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  • 66
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    Microscopy Research and Technique 24 (1993), S. 185-192 
    ISSN: 1059-910X
    Keywords: Electron microscopy ; Epitaxy ; Dislocations ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Transemission electron microscope (TEM) images of dislocations as produced via moiré fringe contrast are simulated using many-beam diffraction theory. The effect of edge dislocations on both parallel and rotational moiré fringe patterns is considered. For the parallel moiré fringe pattern, images of dislocations both perpendicular to the film plane and those inclined to the film plane are produced. The effect of an inclined dislocation is shown to cause a distortion of the dislocation image. Finally, a comparison between predicted and experimentally observed images is made, with the results indicating that threating dislocations in the FeAl/GaAs system have line directions nearly perpendicular to the (001)FeAl/GaAs film plane. © 1993 Wiley-Liss, Inc.
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  • 67
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    Developmental Dynamics 198 (1993), S. 241-253 
    ISSN: 1058-8388
    Keywords: Chick ; Hindbrain ; Rhombomere ; Boundary ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The chick embryo hindbrain is a segmented region of the CNS characterised by repeated blocks of neuroepithelial cells, known as rhombomeres. Individual rhombomeres are polyclonal compartments, defined both by cell lineage restriction and by the restricted expression of development control genes, that later acquire specific patterns of neuronal differentiation and axon outgrowth. The interfaces between adjacent rhombomeres are defined by boundaries across which cells do not move; the boundaries contain specialised cells and are preferentially colonised at early stages of development by extending axons. In this study, routine electron microscopy and high-pressure cryopreservation, a technique that avoids artifacts of chemical fixation, have been used to examine the morphology of rhombomere boundaries through a staged series of chick embryos. We find that the boudary regions contain enlarged extracellular spaces and that these form conduits for axons subsequently extending in the circumferential plane of the hindbrain. Labeling the ventricular surface of the neuroepithelium with DiI crystals in aqueous suspension revealed the morphology of individual cells in the intact neural tube, and demonstrated unusual fan-shaped arrays of cells at the boundaries. These findings contribute further to the evidence that cells at rhombomere boundaries differ from those in rhombomere centres, and leads to hypotheses about both the mechanism of development of the boundaries, and the role they may play in hindbrain patterning. © 1993 Wiley-Liss, Inc.
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  • 68
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    Microscopy Research and Technique 24 (1993), S. 195-213 
    ISSN: 1059-910X
    Keywords: Deafferentation ; Regeneration ; Sensory afferents ; Electron microscopy ; Olfactory receptor cells ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The effects of intranasal zinc sulfate (ZnSO4) irrigation on the morphology of the olfactory epithelium and olfactory bulb were studied in mice with short survival times (as early as 1 day) and with long survival times (up to 593 days) after the irrigation procedure. As in several previous studies, the olfactory epithelium was completely destroyed within a few days after the ZnSO4 treatment. Within 2-4 days, the septum and turbinates were covered by a new, cuboidal epithelium, the cells of which differed significantly from any cells normally seen in the olfactory epithelium. Slowly, over several months, small areas of the olfactory epithelium regenerated in many of the animals.The ultrastructural changes occurring in the olfactory bulb from 1 to 25 days (the reactive stage) were characterized by degenerating olfactory axons and axon terminals, hypertrophy of astroglial cell processes, and proliferation of or extravasation by phagocytic cells. By 25 days after intranasal ZnSO4 irrigation, the number of reactive glial processes and phagocytic cells returned to normal. In some mice with survival times of 150 days or longer, there was reinnervation of small areas of the olfactory bulb by regenerated olfactory axons. These new olfactory axons innervated only superficial glomeruli or the outer portions of deeper glomeruli, but they formed synaptic contacts with mitral/tufted cells and periglomerular cells that did not differ from control animals. These findings were supported by tract-tracing experiments with 3H-amino acids and by behavioral analysis.In summary, the ultrastructural changes observed in the olfactory bulb in this study were not significantly different from those observed after surgical lesions of the olfactory epithelium or nerve. The olfactory bulb, however, never fully recovered; glomeruli remained shrunken (though with normal dendro-dendritic synaptic connections), and there was minimal olfactory axon reinnervation. © 1993 Wiley-Liss, Inc.
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  • 69
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    Microscopy Research and Technique 24 (1993), S. 260-280 
    ISSN: 1059-910X
    Keywords: Neuroanatomy ; Invertebrate brain ; Neurons ; Glial cells ; Olfactory system ; Immunocytochemistry ; Electron microscopy ; Neural development ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Many insects possess a highly developed sense of smell. This paper summarizes the cellular and synaptic organization of the antennal (olfactory) lobe of the insect brain and then reviews morphological and fine-structural aspects of the development of the lobe. Visualization of synapses between classes of neurons identified by physiological, morphological, or transmitter-cytochemical properties has provided insights into arrangements of contacts and their possible roles in information processing. Studies of development have revealed the requirement for afferent axons from the antenna for the formation of olfactory glomeruli, where virtually all of the synapses in the lobe occur, and have suggested the possibility that glial cells play a role in the instructive influence of the axons on their target neurons in the lobe. The findings reviewed in this paper are primarily from one representative hemimetabolous insect, the American cockroach, and one representative holometabolous insect, a hawkmoth, and comparisons are made with vertebrate systems when appropriate. © 1993 Wiley-Liss, Inc.
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  • 70
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    Microscopy Research and Technique 24 (1993), S. 43-66 
    ISSN: 1059-910X
    Keywords: Retina ; Intracellular staining ; Horseradish peroxidase ; Electron microscopy ; Cone ; Horizontal cell ; Bipolar cell ; Amacrine cell ; Ganglion cell ; Neurotransmitter ; Synaptic plasticity ; Spinules ; Rhodamine ; Double labelling ; Postembedding immunohistochemistry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A variety of intracellular recording and staining techniques has been used to establish structure-function and, in some cases, structure-function-neurochemical correlations in fish, turtle, and cat retinae. Cone photoreceptor-horizontal cell connectivity has been studied extensively in the cyprinid fish retina by intracellular staining with horseradish peroxidase (HRP) and subsequent electron microscopy. The available data suggest that horizontal cell dendrites around the ridge of the synaptic ribbon are postsynaptic, whilst finger-like extensions (“spinules”) of lateral dendrites function as inhibitory feedback terminals. An interesting feature of this inter-action is its plasticity: the feedback pathway is suppressed in the dark and becomes potentiated by light adaptation of the retina.Intracellular recordings and stainings of ganglion cells in both turtle and cat retinae have been possible. Prelabelling of ganglion cells by retrograde transport of rhodamine from the tectum allows ganglion cells to be stained under visual control, and their synaptic inputs determined by electron microscopy. Such studies have been extended to double labelling by using autoradiography or postembedding immunohistochemistry to identify the neurotransmitter content of the labelled cell and/or the neurotransmitter(s) converging upon it. It is envisaged that further applications of intracellular staining followed by double- or even triple-labelling will continue to enhance greatly our understanding of the functional architecture of the vertebrate retina. © 1993 Wiley-Liss, Inc.
    Additional Material: 16 Ill.
    Type of Medium: Electronic Resource
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  • 71
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 173-174 
    ISSN: 1059-910X
    Keywords: Porosity ; Mathematical Morphology ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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  • 72
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 366-373 
    ISSN: 1059-910X
    Keywords: Immunogold labeling ; Electron microscopy ; Lung ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Surfactant proteins A, B, and C (SP-A, SP-B, and SP-C) are synthesized in alveolar type II cells. SP-B and SP-C are both synthesized as large precursor molecules that are proteolytically processed to their mature sizes. In a previous immunoelectron microscopic study, we showed that precursor SP-B is processed to its mature size in multivesicular bodies. In the present study, using a specific antibody aginst precursor SP-C, we demonstrate that precursor SP-C is present in the same intracellular compartments of the biosynthetic pathway, i.e., endoplasmic reticulum, Golgi complex, and multivesicular bodies, as precursor SP-B. Since mature SP-C is known to be present in multilamellar bodies, this suggests a biosynthetic routing and site of processing of this protein similar to those of SP-B. Double-labeling experiments using antibodies against SP-A, precursor SP-B, precursor SP-C, and an antibody against HA I, an adaptor protein involved in the budding of transport vesicles from the Golgi complex, showed that the different surfactant proteins traverse and exit the Golgi complex via the same route. The surfactant proteins do not exit the Golgi complex via HA I-positive coated buds or vesicles. These data are in accordance with the concept that SP-A, SP-B, and SP-C are transported together through the same biosynthetic pathway via multivesicular bodies to multilamellar bodies. © 1993 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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