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  • 1
    ISSN: 1435-5922
    Keywords: Key words: hepatic stellate cell ; Ca2+-Mg2+-ATPase ; Ca2+ pump-ATPase ; immunofluorescence ; ultrastructural localization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract: Intracytoplasmic free calcium ions (Ca2+) are maintained at a very low concentration in mammalian tissue by the extrusion of Ca2+ across a steep extracellular Ca2+ gradient, mainly through the activity of plasma membrane Ca2+ pump-ATPase. The present study aimed to identify, by electron cytochemical and electron immunogold methods, the ultrastructural localizations of two types of plasma membrane Ca2+-ATPase; Ca2+-Mg2+-ATPase and Ca2+ pump-ATPase, in hepatic stellate cells. Liver tissues and isolated hepatic stellate cells (HSCs) were studied. The ultrastructural localization of Ca2+-Mg2+-ATPase activity was examined by the electron cytochemical method of Ando. The localization of Ca2+ pump-ATPase was identified by immunofluorescence. The ultrastructural localization of Ca2+ pump-ATPase was identified by the electron immunogold method. The cytochemical reaction products of Ca2+-Mg2+-ATPase activity were localized on the outer (cavity) side of the plasma membrane of caveolae. Immunofluorescence of Ca2+ pump-ATPase was seen as small dots along the cell edge in HSCs. Immunogold particles indicating the presence of Ca2+ pump-ATPase were identified on the inner (cytoplasmic) side of the plasma membrane of caveolae. We localized Ca2+ pump-ATPase on the inner side of the plasma membrane caveolae and Ca2+-Mg2+-ATPase on the outer leaflet of the caveolar plasma membrane in stellate cells, suggesting that Ca2+ pump-ATPase may play a key role in the Ca2+ reflux.
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  • 2
    ISSN: 1437-773X
    Keywords: Key words Hepatic sinusoidal endothelial cell ; Ca2+-ATPase ; Ultrastructural localization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The intracytoplasmic free calcium ion (Ca2+) concentration is maintained at a low level in mammalian tissues by extruding Ca2+ against a high extracellular Ca2+ concentration, mainly through the activity of the plasma membrane Ca2+-ATPase pump. The objective of the present study was to localize the plasma membrane Ca2+-ATPase activity on hepatic sinusoidal endothelial cells (SECs) by electron microscopic cytochemistry. The ultrastructural localization of Ca2+-ATPase activity on ultrathin sections of liver tissue and cultured SEC monolayer was examined by the electron microscopic cytochemical method of Ando (method A: original method) and by our modified method (method B: shortened fixation method). By method A, scanty cytochemical reaction products of Ca2+-ATPase were found in the SECs. By method B, Ca2+-ATPase activity was clearly localized on the outer surface of the plasma membrane of sinusoidal endothelial fenestrae (SEF). Our modification of Ando's method by shortening the incubation time of liver tissue or isolated SEC sections in the substrate allowed clear demonstration of Ca2+-ATPase activity on the SEF membrane. Use of tangential sections of primary cultures of SEC provided excellent localization results. The cytochemically reactive Ca2+-ATPase expressed on the SEF plasma membrane may be involved in regulation of the intracytoplasmic Ca2+ concentration.
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  • 3
    ISSN: 1437-773X
    Keywords: Key words Endothelin ; Endothelin receptor subtypes ; Sinusoidal endothelial cell ; Hepatic stellate cell ; Peptide nucleic acid probe ; Catalyzed signal amplification ; New in situ hybridization method
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Endothelin (ET) has been implicated in the regulation of hepatic microcirculation and development of portal hypertension, but the role of ET in cirrhosis of the liver has not yet been elucidated. We present here the application of peptide nucleic acid (PNA) probes in a fast and sensitive in situ hybridization (ISH) method for localizing the mRNA of endothelin receptor subtypes in formalin-fixed, paraffin-embedded sections of normal and cirrhotic human liver. The results of ISH using synthetic FITC-labeled PNA probes combined with the catalyzed signal amplification (CSA) system were compared with those using the standard detection system. It was indicated that the CSA-ISH protocol is more sensitive for the detection of mRNA target than the standard ISH protocol. Our results with CSA-ISH showed that the expression of mRNA for the endothelin B receptor was significantly upregulated in hepatic sinusoidal lining cells in cirrhotic human liver tissues compared to control normal liver tissue. Therefore, the CSA detection system may facilitate and enhance the use of in situ hybridization protocols, and CSA-ISH will be used as an important diagnostic technique in the future.
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  • 4
    ISSN: 1432-2307
    Keywords: Axon ; Astrocyte ; Synaptophysin ; Glial fibrillary acidic protein ; Foamy spheroid body
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Foamy spheroid bodies (FSBs) are described, as newly identified pathological structures occurring in human brain. FSBs favoured the substantia nigra pars reticulata (SNPR) and/or globus pallidus (GP) in degenerative conditions especially postencephalitic parkinsonism, progressive supranuclear palsy, pallido-nigro-luysial atrophy and multiple system atrophy. No FSBs were observed anywhere in the presence of substantia nigra pars compacta (SNPC) degeneration, such as occurs in idiopathic Parkinson's disease, or luysio-pallidal system degeneration, such as found in dentato-rubro-pallidoluysial atrophy or Joseph's disease. FSBs were also occasionally identified in the substantia nigra (SN) and/or GP of aged persons. In addition to SN and GP lesions, FSBs were seen in diffuse axonal lesions of long fibre tracts (the corpus callosum, the superior cerebellar peduncle) after non-missile head injuries, and in peri-infarct lesions. Under the light microscope, FSBs appear as slightly eosinophilic, foamy and nearly round objects with vague outlines, measuring approximately 10–50 μm in diameter. Some FSBs contain coarse, eosinophilic clusters at their periphery. FSB stained black when stained by the Gallyas silver method. Some FSBs were immunohistochemically positive for synaptophysin and 68 kDa neurofilament. Glial fibrillary acidic proteinpositive fibres were observed alongside and/or inside some FSBs. Electron microscopically, FSBs were found to consist of collections of neuritic debris containing a variety of dense bodies and a small number of both mitochondria and neurofilaments. Some such collections were surrounded by astrocytic processes. These findings strongly suggest that FSBs are collections of small axonal debris destined for removal by astrocytes in due course. A variety of factors (degeneration of the SNPR and/or the GP, injury, infarction, ageing) seemed to be responsible for the histogenesis of FSBs.
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  • 5
    ISSN: 1432-0533
    Keywords: Multiple system atrophy ; Oligodendroglial cytoplasmic inclusions ; αB-crystallin ; Ubiquitin ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Oligodendroglial cytoplasmic inclusions (OCI) in multiple system atrophy were investigated immunocytochemically and ultrastructurally. Among the 17 cases examined, 16 had OCIs. Almost all OCIs were positive for both αB-crystallin and ubiquitin. The antibodies against tubulin, paired helical filament and tau stained OCIs to various extents. Ultrastructurally OCIs consisted of meshworks of granule-associated filaments about 25 nm in diameter that sometimes formed flame-shaped tangle-like structures. Immunoelectron microscopy showed that an epitope of αB-crystallin was located on the granule-associated filaments composing OCIs. Our studies further support a cooperative role of αB-crystallin, ubiquitin and cytoskeletal protein in the formation of some types of intracytoplasmic inclusions.
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  • 6
    ISSN: 1432-0533
    Keywords: Key words Amyotrophic lateral sclerosis ; Morphometry ; Multiple system atrophy ; Parkinson's ; disease ; Unmyelinated nerve fibres
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Quantitative changes in unmyelinated nerve fibers (UMNFs) of sural nerves in patients of amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD) and multiple system atrophy (MSA) were evaluated using autopsy materials whose pathological diagnosis had been confirmed by careful postmortem examinations. Ordinary ALS cases demonstrated no involvement in cutaneous UMNFs; however, the patients with long survival due to the application of ventilatory support showed bimodality in UMNF diameter histograms, and a patient with involvement of systems other than motor pathways showed an abnormal value in two indices: a low percentage of subunits containing axon(s) and a high mean number of Schwann cell profiles per axon. A significant reduction of the mean value of UMNF density (21%) was found in PD patients. Because the density of myelinated nerve fibers did not show any significant decrease as compared with age-matched controls, the change of nerve fibers in peripheral nervous system was considered to be confined to UMNFs in PD. Elderly PD cases showed enhanced changes in the ageing process, as expressed by the two indices described above. In MSA, the mean value of UMNF density was significantly decreased (23%), and this decrease almost paralleled that of myelinated nerve fiber density. Abnormal values for the two indices described above were found and two out of four cases demonstrated bimodality in the diameter histogram of UMNFs. Unlike MSA, ALS and PD have not been included in the disorders with cutaneous UMNF involvement. Our results supply the first evidence of morphological changes in cutaneous UMNFs in PD cases. In ordinary ALS cases, the emergence of such morphological changes is suggested in cases with long survival.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 96 (1998), S. 52-56 
    ISSN: 1432-0533
    Keywords: Key words Calcium ion ; Tyrosine kinase ; Golgi ; apparatus ; Protein traffic ; Axonal transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The expression of protein kinase C (PKC), a calcium- and phospholipid-dependent signaling molecule, was studied immunohistochemically in the spinal motor neurons of cases of sporadic amyotrophic lateral sclerosis (SALS). In the normal spinal cord, intense PKC immunoreactivity was found in subsets of large motor neurons. PKC immunoreactivity was markedly decreased in the spinal motor neurons of SALS. The result suggests that down-regulation of PKC is associated with the degeneration of spinal motor neurons in SALS.
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  • 8
    ISSN: 1432-0533
    Keywords: Key words Macroglobulinemia ; Neuropathy ; Blood-nerve barrier ; HNK-1 epitope ; Endothelial cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We report the pathologic findings in a patient with sensorimotor neuropathy associated with Waldenström’s macroglobulinemia, particularly in relation to blood-nerve barrier defects. The monoclonal IgM was of κ type and possessed anti-HNK-1 activity. A sural nerve biopsy specimen revealed severe loss of myelinated and unmyelinated nerve fibers and gaps between adjacent endothelial cells of small endoneurial vessels. Postmortem findings 3 years later included severe loss of myelinated nerve fibers and diffuse infiltration by lymphoplasmacytic B cells throughout the peripheral nervous system, sparing the central nervous system. Findings in this case suggest an immune attack against endoneurial endothelial cells with permeation of IgM into peripheral nerve tissue.
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  • 9
    ISSN: 1432-0533
    Keywords: Key words Secondary degeneration ; Neurofibrillary ; tangle ; Progressive supranuclear palsy ; Substantia ; nigra ; Dentate nucleus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A case of hemispheric infarction involving the territory of the right middle cerebral artery and the thalamus showed conspicuous asymmetric degeneration in the substantia nigra, red nuclei, inferior olivary nuclei and dentate nuclei with concomitant changes of progressive supranuclear palsy (PSP). The right substantia nigra and red nucleus showed loss of neurons and proliferation of astrocytes. The right olivary nucleus was hypertrophic, while the neuronal loss and astrocytosis in the dentate nucleus were predominant on the contralateral side. Modified Gallyas-Braak staining revealed the extensive distribution of neurofibrillary tangles (NFTs), threads and intraglial argyrophilic structures in the globus pallidus, subthalamic nuclei, cerebral cortex and dentate nuclei, as well as in the affected brain stem nuclei, with a distinct predominance on the affected side. In this case, the one-sided predominance of the extended degeneration in these brain stem and cerebellar areas is considered, in addition to the PSP changes, to be due to secondary retrograde degeneration via the nigrostriatal and dentato-rubro-thalamic pathways following the hemispheric infarction, and to also be the result of disruption of the dentato-olivary fiber connections. In addition, because of the predominant distribution of NFTs on the more degenerated side, it is surmised that the formation of NFTs may be accelerated by secondary degeneration.
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  • 10
    ISSN: 1432-0533
    Keywords: Key words Amyotrophic lateral sclerosis ; SOD1 gene ; Posterior column ; Lewy-body-like inclusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A 65-year-old man with familial amyotrophic lateral sclerosis (ALS) with posterior column involvement showed fairly slow progression of the illness and lived with the aid of a respirator for 12 years. Neuropathological examinations showed simultaneous involvement of the pyramidal tract and lower motor neurons as well as degeneration in the Clarke’s nucleus- spinocerebellar tract- middle root zone of the posterior column, the pallido-luysian system, the medullary reticular formation, and widespread anterolateral columns of the spinal cord. However, the patient had no Lewy-body-like hyaline inclusions, which are characteristic features of this form of familial ALS. Moreover, no abnormalities were found in his SOD1 cDNA sequences. There seem to be certain heterogeneities in familial ALS with posterior column involvement, and SOD1 gene abnormalities may be involved in the pathomechanism in rapidly progressing ALS, in which there are Lewy-body-like hyaline inclusions.
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