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Biomedical subjects

C Ide

Publications and source records attributed to C Ide.

At least 37 records · Page 2Linked to original sources
PubMed2000

Choroid plexus ependymal cells enhance neurite outgrowth from dorsal root ganglion neurons in vitro.

The epithelial cells of the choroid plexus are a continuation of the ventricular ependymal cells and are regarded as modified ependymal cells. The present study was carried out to determine the influence of choroid plexus ependymal cells (CPECs) on axonal growth in vitro. Choroid plexuses were dissected from the fourth ventricle of postnatal day-1-10 mice, mechanically dissociated, and plated in fibronectin-coated culture dishes. CPECs had spread into monolayers with few endothelial cells in 3-week cultures. Some macrophages were scattered on the monolayer of CPECs. Dorsal root ganglia (DRG) were excised from mouse fetuses of 14-day gestation, dissociated with trypsin and cocultured on the CPEC monolayers. For comparison, dissociated DRG neurons were cocultured on astrocyte monolayers or cultured on laminin-coated plates. After 4.5 h culturing, the cultures were fixed and immunohistochemically double-stained for neurites and CPECs using antibodies against beta-tubulin III and S-100 beta, respectively. It was demonstrated that neurons extended many long neurites with elaborate branching on the surface of S-100-stained CPECs. In contrast, DRG neurons cultured on the astrocytes and on the laminin-coated plates had much shorter primary neurites with fewer branches than those cultured on the CPECs. The total length of neurites including primary neurites and their branches, of a single DRG neuron was 285 +/- 14, 395 +/- 15 and 565 +/- 12 microM on the laminin-coated plates, on astrocytes and on CPECs, respectively. Scanning electron microscopy revealed extension of neurites with well-developed growth cones on the ependymal cells. These results suggest that CPECs have a great capacity to promote neurite outgrowth from DRG neurons in vitro.

Animals↗
PubMed2000

Ultrastructural characteristics and synaptophysin immunohistochemistry of regenerating nerve growth cones following traumatic injury to rat peripheral nerve.

Growth cones of regenerating nerves, following crush injury to the rat peripheral nerve system, were studied by electron microscopy and synaptophysin. Localization in the growth cones was revealed by immunohistochemistry at the ultrastructural level. Many regenerating growth cones grew along the Schwann-cell basal laminae tubes at the crushed site. These cones revealed an abundance of organelles, such as heterogenous vesicles and many mitochondria, and a scarcity of cytoskeletons, including microtubules and neurofilaments in the cytoplasm. The periphery of the growth cones (corresponding to the lamellipodia or filopodia of cultured neurons) contained rich electron-dense filamentous materials. Cellular protrusions, such as filopodia, were rarely seen. These growth cones exhibited intense immunoreactivity for synaptophysin by light microscopy. Immunoelectron microscopy demonstrated that immunoreactivity was distributed diffusely in the cytoplasm.

Animals↗
PubMed2000

Positron emission tomography with fluorodeoxyglucose for suspected head and neck tumor recurrence in the symptomatic patient.

OBJECTIVE: To analyze the impact of positron emission tomography with fluorodeoxyglucose (FDG-PET) in the treatment of patients suspected of having head and neck cancer recurrence. STUDY DESIGN: Prospective and consecutive inclusion of 44 patients presenting with clinical symptoms suggestive of head and neck tumor recurrence. METHODS: FDG-PET was compared with combined computed tomography (CT) plus magnetic resonance imaging (MRI) procedures for the differential diagnosis between tumor recurrence and benign post-therapeutic changes. For FDG-PET, the potential additional value of semiquantitative indexes was studied. The impact on patient treatment (i.e., their ability to accurately select patients for panendoscopic exploration) was analyzed retrospectively for both CT+MRI and PET workups. RESULTS: The diagnostic accuracy was found higher for PET than for combined CT+MRI: sensitivity ranged from 96% to 73%, specificity from 61% to 50%, and accuracy from 81% to 64% for PET and CT+MRI, respectively. The accuracy of FDG-PET was the highest (94%) in patients included more than 12 weeks after the end of therapy. In 15 discordant cases, PET was correct in 11 and CT+MRI in 4. Patient selection for panendoscopic exploration and biopsy was correct in 79% and 50% of patients with FDG-PET and CT+MRI, respectively. Quantification of FDG uptake had no additional value over visual analysis alone, although we found that a SUVlbm (standardized uptake value corrected for lean body mass) threshold of 3 could be helpful in patients scanned less than 12 weeks after the end of therapy. CONCLUSION: FDG-PET has a major additional diagnostic value to CT+MRI for the evaluation of the symptomatic patient suspected of having head and neck cancer recurrence. PET could have a direct impact on management by correctly selecting patients in whom a panendoscopic exploration with biopsy is indicated.

Adult↗
PubMed2000

Distinct aggregation of beta- and gamma-chains of the high-affinity IgE receptor on cross-linking.

The high-affinity IgE receptor (FcepsilonRI) on mast cells and basophils consists of a ligand-binding alpha-chain and two kinds of signaling chains, a beta-chain and disulfide-linked homodimeric gamma-chains. Crosslinking by multivalent antigen results in the aggregation of the bound IgE/alpha-chain complexes at the cell surface, triggering cell activation, and subsequent internalization through coated pits. However, the precise topographical alterations of the signaling beta- and gamma-chains during stimulation remain unclarified despite their importance in ligand binding/signaling coupling. Here we describe the dynamics of FcepsilonRI subunit distribution in rat basophilic leukemia cells during stimulation as revealed by immunofluorescence and immunogold electron microscopy. Immunolocalization of beta- and gamma-chains was homogeneously distributed on the cell surfaces before stimulation, while crosslinking with multivalent antigen, which elicited optimal degranulation, caused a distinct aggregation of these signaling chains on the cell membrane. Moreover, only gamma- but not beta-chains were aggregated during the stimulation that evoked suboptimal secretion. These findings suggest that high-affinity IgE receptor beta- and gamma-chains do not co-aggregate but for the most part form homogenous aggregates of beta-chains or gamma-chains after crosslinking.

Animals↗
PubMed2000

Incorporation and differentiation of hippocampus-derived neural stem cells transplanted in injured adult rat retina.

PURPOSE: In a previous study it has been shown that adult rat hippocampus-derived neural stem cells can be successfully transplanted into neonatal retinas, where they differentiate into neurons and glia, but they cannot be transplanted into adult retinas. In the current study, the effect of mechanical injury to the adult retina on the survival and differentiation of the grafted hippocampal stem cells was determined. METHODS: Mechanical injury was induced in the adult rat retina by a hooked needle. A cell suspension (containing 90,000 neural stem cells) was slowly injected into the vitreous space. The specimens were processed for immunohistochemical studies at 1, 2, and 4 weeks after the transplantation. RESULTS: In the best case, incorporation of grafted stem cells was seen in 50% of the injured retinas. Most of these cells located from the ganglion cell layer through the inner nuclear layer close to the injury site. Immunohistochemically, at 1 week, more than half of the grafted cells expressed nestin. At 4 weeks, some grafted cells showed immunoreactivity for microtubule-associated protein (MAP) 2ab, MAP5, and glial fibrillary acidic protein (GFAP), suggesting progress in differentiation into cells of neuronal and astroglial lineages. However, they showed no immunoreactivity for HPC-1, calbindin, and rhodopsin, which suggests that they did not differentiate into mature retinal neurons. Immunoelectron microscopy revealed the formation of synapse-like structures between graft and host cells. CONCLUSIONS: By the manipulation of mechanical injury, the incorporation and subsequent differentiation of the grafted stem cells into neuronal and glial lineage, including the formation of synapse-like structures, can be achieved, even in the adult rat retina.

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PubMed1999

Distribution of synaptosomal-associated protein 25 in nerve growth cones and reduction of neurite outgrowth by botulinum neurotoxin A without altering growth cone morphology in dorsal root ganglion neurons and PC-12 cells.

Synaptosomal-associated protein 25 has been regarded as one of the target-associated soluble N-ethylmaleimide-sensitive fusion attachment protein receptors essential for exocytosis of vesicles in synapses. We have previously reported that cleavage of syntaxin, which is another target-associated soluble N-ethylmaleimide-sensitive fusion attachment protein receptor, with botulinum neurotoxin C1 resulted in inhibition of neurite extension and morphological changes including growth cone collapse and large vacuole formation. As an attempt to explore the mechanism of growth cone extension, we examined the ultrastructural localization of synaptosomal-associated protein 25 in growth cones with or without treatment of botulinum neurotoxin A, which cleaves synaptosomal-associated protein 25. In dorsal root ganglion neurons, light microscopy demonstrated synaptosomal-associated protein 25 immunoreactivity throughout the neurons, including the cell bodies, neurites and growth cones. Using electron microscopy, gold signals immunoreactive for synaptosomal-associated protein 25 were identified diffusely in the cytoplasm of the growth cones. In contrast, in PC-12 cells, a large number of gold signals were localized on the plasma membranes. High levels of signal were also found in the cytoplasm in the central region of the growth cones. We also confirmed that botulinum neurotoxin A treatment reduced neurite extension by about 50%. However, both in dorsal root ganglion neurons and in PC-12 cells we found no differences in the ultrastructure nor in the localization of synaptosomal-associated protein 25 between growth cones with and without toxin treatment. These results indicate that cleavage of synaptosomal-associated protein 25 inhibits growth cone extension in a manner different than that of syntaxin cleavage. The results of this study suggest the possibility that synaptosomal-associated protein 25 is involved in growth cone extension through a process independent of vesicle fusion.

Animals↗
PubMed1999

Nerve regeneration through biodegradable gelatin conduits in mice.

OBJECTIVE: To test if fabricated gelatin conduits can be used to bridge nerve inter-stump gaps and support regeneration. DESIGN: Experimental laboratory study. SETTING: Department of Anatomy and Neurobiology, Graduate School of Medicine, Kyoto University, Japan. SUBJECTS: Twenty-four adult mice. INTERVENTION: Mouse's sciatic nerve was resected and both proximal and distal nerve stumps sutured into each end of a gelatin conduit, to bridge a 7-mm gap. MAIN OUTCOME MEASURES: Nerve regeneration. RESULTS: At one week post-implantation, a scaffolding fibrin matrix containing few mononuclear cells formed inside the conduit. At three weeks, a well regenerated nerve composed of myelinated and unmyelinated axons, associated Schwann cells and surrounding perineurial sheath bridged the gap. CONCLUSION: Biodegradable gelatin conduits direct and support nerve regeneration and are therefore promising tools for use in entubulization repair of nerve defects.

Animals↗
PubMed1998

Regeneration of dorsal column axons after spinal cord injury in young rats.

In contrast to previous reports denying the occurrence of axonal regeneration of the dorsal column (DC) projections, here we demonstrate for the first time that marked regeneration occurs spontaneously after transection in infant rats. Transection was made sharply so as to produce edema-free lesions without subsequent formation of either scars or cysts. Transganglionic labeling of axons revealed that regenerated axons ascended in the normal tract in a manner similar to normal projections as a tightly-packed fasciculus and terminated densely in the nucleus gracilis. The present study indicates that failure of regeneration of DC axons is due to neither intrinsic deficiency of regrowth potential nor globally-inhospitable axonal environment but rather the local conditions of the lesion site.

Animals↗
PubMed1998

Localization of tyrosine-phosphorylated proteins in cultured mouse dorsal root ganglion neurons.

The present study, using confocal laser scanning microscopy and immunoelectron microscopy, examined the intracellular localization of tyrosine-phosphorylated proteins in cultured mouse dorsal root ganglion neurons with special reference to their growth cones. The growth cone is the specialized structure formed at the growing tip of the axon; characteristically highly motile with filopodia on the surface, it is responsible for the extension and guidance of the neurites to the appropriate targets during nerve regeneration. It has been suggested that protein-tyrosine phosphorylation plays an important role in the intracellular signal transduction that regulates the extension and motility of growth cones. By fluorescence immunocytochemistry, phosphotyrosine immunoreactivity was found in the growth cones and neurites. Some of the filopodia exhibited strong immunoreactivity at their tips. By immunoelectron microscopy, a large number of immunogold particles (gold particles conjugated to the secondary antibody) were seen to be distributed in the cytoplasm and some were observed on the plasma membrane in the growth cones, whereas in the neurites the density of immunogold particles was the same in the axoplasm as on the plasma membranes. These findings suggest that in the growth cones phosphotyrosines might mainly be involved in intracellular signaling for maintaining their high motility whereas in the neurites they might mostly be associated with the receptor proteins at the plasma membrane for adhesion as well as for growth of neurites. Thus, tyrosine phosphorylation might contribute to different functions for growth cones and neurites.

Animals↗
PubMed1998

Long acellular nerve transplants for allogeneic grafting and the effects of basic fibroblast growth factor on the growth of regenerating axons in dogs: a preliminary report.

Sciatic nerves were excised from 3 beagle dogs about 5 h after their sacrifice, treated three times by freezing and thawing, and stored in physiological saline for 3 months at -20 degrees C until used. Nerve segments 5 cm in length prepared from these stored nerves were transplanted to the common peroneal nerve in the right hindlimb of beagle dogs. Sixteen beagle dogs in total were used, in four treatment groups of two pairs each studied at 1 and 3 months. Five-hundred microliters basic fibroblast growth factor (bFGF) of two different concentrations (10 micrograms/300 microliters and 100 micrograms/300 microliters) which were impregnated in 0.5 ml gelatin hydrogels was applied around the sutured allografts. Autografting was also done in 4 beagle dogs, with no bFGF application. One month after the grafting, no regenerating nerves extended beyond the middle of the transplant in any of the allografts, except in the autografts in which a number of regenerated (myelinated) axons were present. Three months after the grafting, an abundance of myelinated axons was found at the middle of the graft: the numbers of axons per 10(4) micron 2 were 22.6 in the autografts and 10.6, 10.4 and 19.2 in the allografts treated with no bFGF, low-dose bFGF, and high-dose bFGF, respectively. Regenerating axons extended into the host nerve: the numbers of myelinated axons at the level 1.5 cm distal to the distal suture were 35.7, 0.9, 3.8, and 12.1 per 10(4) micron 2 in the above respective order. Although it was inferior in quality to the autograft, peripheral nerve regeneration was extensive in the distal nerve using freeze-thawed and bFGF-treated allografts at 3 months. Electromyography showed that the peroneus longus muscle responded to the electrical stimuli given at the site proximal to the transplant in all four groups. These data indicate that a 5-cm acellular nerve segment containing Schwann cell basal laminae can be used successfully as an allograft without any immunosuppressants and that exogenously applied bFGF can improve nerve regeneration by enhancing the growth of regenerating axons.

Animals↗
PubMed1998

Effects of prelesioned peripheral nerve graft on nerve regeneration in the rat spinal cord.

OBJECTIVE: The aim of this study was to examine the effects of prelesioned peripheral nerve grafts on central nerve regeneration compared with the freshly transected peripheral nerve grafts in the dorsal funiculus of the rat spinal cord. METHODS: The experimental paradigm consisted of ligating the common peroneal nerve at the midthigh level for 7 days, while the adjacent tibial nerve was left intact. Numerous Schwann cells appeared accompanying regenerating axons in the proximal stump of the ligated nerve. The proximal stumps of the ligated (prelesioned) common peroneal nerve and the intact (untreated) tibial nerve were excised as one tissue block and autografted into the dorsal funiculi of the upper cervical cord. The graft was placed so that the prelesioned common peroneal nerve was positioned on the left dorsal funiculus and the untreated tibial nerve was positioned to the right of the midsagittal plane. Nerve regeneration was examined by light and transmission electron microscopy 1 to 16 weeks after grafting, comparing the effectiveness of prelesioned and untreated nerve grafts. RESULTS: Numerous regenerating axons were observed in the caudal border of both grafts 1 to 2 weeks after grafting. Astrocyte proliferation was suppressed in the prelesioned grafts compared to the untreated grafts. Four to 16 weeks later, the number of regenerating axons was approximately 10-fold as large in the prelesioned grafts as in the untreated grafts. The regenerating axons were myelinated by Schwann cells. Astrocytic glial scar formation was inconspicuous in the prelesioned grafts, whereas it was prominent in the untreated grafts. Schwann cells were contiguous with astrocytes along regenerating axons, forming a continuous conduit from the central to peripheral nerve microenvironments for the outgrowth of regenerating axons. CONCLUSION: The prelesioned peripheral nerve graft is more effective than the untreated graft in suppressing astrocytic scar formation and in supporting the outgrowth of regenerating axons in the dorsal funiculus of rat spinal cord.

Animals↗
PubMed1998

Unusual occipitoatlantal fracture dissociation with no neurological impairment. Case report.

The authors describe an unusual case of a complex traumatic fracture-dissociation injury of the craniovertebral junction, which the patient survived with no neurological damage. This case featured the rare combination of an avulsion of both the right occipital condyle and clivus and a fracture of the left lateral mass of the atlas. Because of the craniocervical ligament injury and the slight anterior occipitoatlantal dislocation, the lesion was considered to be unstable and was treated successfully with a cervical collar. The authors emphasize that thin-slice computerized tomography scanning with multiplanar reconstructions is essential to visualize these fractures, whereas magnetic resonance imaging is useful to assess soft tissues.

Adult↗
PubMed1998

An early MR observation of carotid involvement by retropharyngeal abscess.

We report early carotid involvement by retropharyngeal abscess in a 4-year-old boy. MR imaging showed enhancement of the wall and narrowing of the lumen of the internal carotid artery, which were thought to reflect spasm and/or arteritis. Prompt treatment may have prevented hemorrhagic and neurologic complications.

Carotid Arteries↗
PubMed1997

The Ras target AF-6 interacts with ZO-1 and serves as a peripheral component of tight junctions in epithelial cells.

The dynamic rearrangement of cell-cell junctions such as tight junctions and adherens junctions is a critical step in various cellular processes, including establishment of epithelial cell polarity and developmental patterning. Tight junctions are mediated by molecules such as occludin and its associated ZO-1 and ZO-2, and adherens junctions are mediated by adhesion molecules such as cadherin and its associated catenins. The transformation of epithelial cells by activated Ras results in the perturbation of cell-cell contacts. We previously identified the ALL-1 fusion partner from chromosome 6 (AF-6) as a Ras target. AF-6 has the PDZ domain, which is thought to localize AF-6 at the specialized sites of plasma membranes such as cell-cell contact sites. We investigated roles of Ras and AF-6 in the regulation of cell-cell contacts and found that AF-6 accumulated at the cell-cell contact sites of polarized MDCKII epithelial cells and had a distribution similar to that of ZO-1 but somewhat different from those of catenins. Immunoelectron microscopy revealed a close association between AF-6 and ZO-1 at the tight junctions of MDCKII cells. Native and recombinant AF-6 interacted with ZO-1 in vitro. ZO-1 interacted with the Ras-binding domain of AF-6, and this interaction was inhibited by activated Ras. AF-6 accumulated with ZO-1 at the cell-cell contact sites in cells lacking tight junctions such as Rat1 fibroblasts and PC12 rat pheochromocytoma cells. The overexpression of activated Ras in Rat1 cells resulted in the perturbation of cell-cell contacts, followed by a decrease of the accumulation of AF-6 and ZO-1 at the cell surface. These results indicate that AF-6 serves as one of the peripheral components of tight junctions in epithelial cells and cell-cell adhesions in nonepithelial cells, and that AF-6 may participate in the regulation of cell-cell contacts, including tight junctions, via direct interaction with ZO-1 downstream of Ras.

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