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Biomedical subjects

K Tohyama

Publications and source records attributed to K Tohyama.

At least 91 records · Page 5Linked to original sources

Nerve regeneration through the cryoinjured allogeneic nerve graft in the rabbit.

To examine whether the 3-4-cm-long allogeneic basal lamina tubes of Schwann cells serve as conduits for regenerating axons in rabbits, allogeneic saphenous nerve, which had been predenervated and pretreated by freezing, were transplanted from Japanese White rabbits (JW) to New Zealand White rabbits (NW). Animals were killed 1, 2, 6, 8, and 14 weeks after transplantation, and the cytology at the mid-portion of the grafts was examined by electron microscopy. The distal portion of the host saphenous nerves was also examined 14 weeks after grafting. Myelin sheath debris was phagocytosed by macrophages, while the basal lamina of Schwann cells were left intact in the form of tubes. Regenerating axons were first found in such basal lamina tubes 2 weeks after grafting, and gradually increased in number. Host Schwann cells accompanied the regenerating axons behind their growing tips, separating them into individual fibers and forming thin myelin sheaths on thick axons by 6 weeks after grafting. Regenerating nerves were divided into small compartments by new perineurial cells. Newly formed blood vessels were situated outside the compartment 8 weeks after grafting. The percentage of myelinated fibers in the regenerating nerves was roughly 10% at 8 weeks and 30% at 14 weeks after grafting. The diameter of the regenerating axons, both myelinated and unmyelinated, was less than that of normal axons at all the stages examined. Numerous regenerating axons, some of which were fully myelinated, were found at the site 10 mm distal to the distal end of the graft 14 weeks after grafting.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Allogeneic nerve grafts in the rat, with special reference to the role of Schwann cell basal laminae in nerve regeneration.

The role of basal laminae as conduits for regenerating axons in an allogeneic graft was examined by transplanting a 3 cm long segment of the sciatic nerve from the Brown Norway to the Fischer 344 strain of rat. These strains are not histocompatible with each other. In order to compare the nerve regeneration in variously treated grafts, three different types of graft were employed: non-treated (NT), predenervated (PD), and predenervated plus freeze-treated (PDC) grafts. The cytology of nerve regeneration through these grafts was examined by electron microscopy at four, seven, 14, 30 and 60 days after grafting. In the PDC graft, in which Schwann cells were dead on grafting, basal laminae were well preserved in the form of tubes after Schwann cells and myelin sheaths had been removed at seven days after grafting. Regenerating axons accompanied by immature host Schwann cells grew out through such basal lamina tubes in the same fashion as observed in our previous studies. By day 14, axons extended as far as the middle of the graft. In the proximal part they were separated into individual fibres and even thinly myelinated by Schwann cells. On the other hand, in the NT and PD grafts in which Schwann cells were alive on grafting, most Schwann cells and myelin sheaths appeared to undergo autolytic degeneration by day 14, while Schwann cell basal laminae were left almost intact in the form of tubes. A few regenerating axons were seen associated with Schwann cells in the proximal portion by day seven. It is probable that host Schwann cells moved into the graft after donor cells had been degraded. Schwann cell basal laminae tended to be damaged at the site of extensive lymphoid cell infiltration. By day 30, regenerating axons had arrived at the distal end of the graft in all three types of graft: in the PDC graft thick axons were fully myelinated, whereas in the PD graft they were only occasionally myelinated and in the NT graft most axons were still surrounded by common Schwann cells. By 60 days after grafting, regenerating axons were well myelinated in the host nerve as observed 1 cm distal to the apposition site in all the three types of graft. These findings show that Schwann cell basal laminae can serve as pathways (most efficiently in the PDC graft) for regenerating axons in a 3 cm long allograft in the rat.

Animals↗

Effects of DM-9384, a pyrrolidone derivative, on alcohol- and chlordiazepoxide-induced amnesia in mice.

The effects of N-(2,6-dimethyl-phenyl)-2-(2-oxo-1-pyrrolidinyl) acetamide (DM-9384), a new pyrrolidone derivative, were investigated on ethanol- and chlordiazepoxide (CDP)-induced amnesia animal model using the passive avoidance task in comparison with aniracetam, another pyrrolidone derivative. Pretraining administration of DM-9384 attenuated ethanol- and CDP-induced amnesia, whereas aniracetam failed to do so. The effects of DM-9384 on CDP-induced amnesia were antagonized by bicuculline, a GABAA receptor antagonist, but not by scopolamine, a muscarinic acetylcholine receptor antagonist and flumazenil, a benzodiazepine receptor antagonist. These results suggest that DM-9384 attenuates CDP-induced amnesia by interacting with the GABAergic neuronal system.

Amnesia↗

Myelodysplastic syndrome (MDS)-associated inhibitory activity on haemopoietic progenitor cells.

We studied MDS-associated inhibitory activity, which inhibited colony formation in vitro of granulocyte-macrophage progenitors (CFU-GM). Macrophages obtained from MDS bone marrow mononuclear cells (BM-MNC) when pretreated with granulocyte-macrophage colony stimulating factor (GM-CSF) suppressed the growth of normal CFU-GM. These macrophages were designated as 'MDS-derived inhibitory macrophages'. Media conditioned by MDS-derived inhibitory macrophages (MDS-CM) also suppressed the growth of normal CFU-GM. In the MDS-CM, high levels of prostaglandin E2 (PGE2) and ferritin were found. However, MDS-CM did not contain detectable levels of tumour necrosis factor (TNF) or gamma-interferon (gamma-IFN). Antiserum against human placental ferritin and/or against PGE2 blocked the haemopoietic inhibitory activity to some extent. These results suggest that inhibitory macrophages may be responsible for the suppression of granulopoiesis in patients with MDS and that the suppression may be mediated by soluble factors including PGE2 and ferritin.

Adult↗

Effects of benzodiazepines on passive avoidance response and latent learning in mice: relationship to benzodiazepine receptors and the cholinergic neuronal system.

The effects of benzodiazepines on learning and memory were investigated, using passive avoidance and latent learning tasks, with particular attention being paid to the possible involvement of benzodiazepine receptors and the cholinergic neuronal system. Benzodiazepines such as diazepam, nitrazepam and chlordiazepoxide (CDP) impaired the passive avoidance response when administered before training, but not when administered immediately after training or before the retention test. CDP also impaired latent learning in the water finding task. State-dependent learning was not observed with CDP at the dose used. A benzodiazepine inverse agonist, Ro 15-4513, and a benzodiazepine antagonist, Ro 15-1788, completely and partially reversed, respectively, the disruptive effects of CDP on learning and memory at the doses which did not enhance learning and memory. The disruptive effects of CDP on learning and memory were partially antagonized by a choline esterase inhibitor, physostigmine, and by a blocker for muscarinic acetylcholine receptors, scopolamine, at the doses which increase acetylcholine release. These results suggest that benzodiazepines induce disruptive effects on learning and memory through benzodiazepine receptors, and that benzodiazepine-induced impairment of learning and memory is, at least in part, the result of the dysfunction of the cholinergic neuronal system.

Animals↗

Regenerating axons and their growth cones observed by scanning electron microscopy.

A predenervated sciatic nerve segment, which had been treated by repeated freezing and thawing to kill Schwann cells, was grafted to the original sciatic nerve in the rat. Three to five days later, the graft was chemically fixed and treated by KOH-collagenase digestion, a treatment which selectively removes almost all non-cellular elements including the collagen fibrils and basal laminae from the tissue, thus making it possible to observe regenerating axons by scanning electron microscopy. Debris of degraded Schwann cells and myelin sheaths remained in the form of "columns", and several thick (2-3 microns in diameter) and thin (less than 1 micron in diameter) axons ran singly or in bundles on such "cell debris columns." Thick axons have an almost straight contour, while there were various swellings at intervals along the thin axons. In most cases, the growing tips of regenerating axons were swollen as growth cones ranging from 2 microns to 5 microns in diameter. Growth cones exhibited fusiform to polygonal variations in structure and had only a few filopodial processes on the surface.

Animals↗

Effects of metaphit on phencyclidine and serotonin2 receptors.

We have investigated whether metaphit, a derivative of phencyclidine (PCP) which irreversibly binds to a population of PCP receptor sites in rat brain, blocks PCP-induced head-twitch response which is produced through serotonin2 (5-HT2) receptors, and also whether metaphit decreases the capacity of 5-HT2 receptors. Metaphit (1 mumol/rat) had decreased the intensity of PCP-induced head-twitch response and had depleted both PCP and 5-HT2 receptors by 24 h after administration, but it failed to block 5-HT agonist 5-methoxy-N,N-dimethyltryptamine-induced 5-HT1A receptor-dependent behaviors. These results reconfirmed our hypothesis that PCP and 5-HT2 receptors may have very similar binding sites.

Animals↗

Cooperative effects of gamma-interferon and 1 alpha, 25-dihydroxyvitamin D3 on in vitro differentiation of the blast cells of RAEB and RAEB-T.

We added recombinant human gamma-interferon (gamma-IFN) and 1 alpha, 25-dihydroxyvitamin D3 (1 alpha, 25 (OH)2D3) to the bone marrow cells from six patients with RAEB or RAEB-T in liquid suspension cultures. After cultivation for 7 to 9 days, numerical, morphological and functional changes of the cells were assessed. gamma-IFN and 1 alpha, 25 (OH)2D3 additively suppressed cell growth, especially the number of blast cells decreased. The expression of alpha-naphthylbutyrate esterase (NBE) activity appeared to be promoted but that of naphthol AS-D chloroacetate esterase (NAE) activity was apparently suppressed by the addition of gamma-IFN and/or 1 alpha, 25 (OH)2D3. The percentage of NBT reduction-positive cells and latex-phagocytizing cells was only slightly increased by both agents. These results indicate that gamma-IFN and 1 alpha, 25 (OH)2D3 cooperate to induce monocytoid differentiation of the patients' blast cells. Combination therapy with both agents merits further study.

Adult↗

Prognostic implication of sequential bone marrow cultures in the myelodysplastic syndromes.

Twenty-five patients with newly-diagnosed myelodysplastic syndromes were studied by the clonal culture method at least three times during the clinical course. Clinical outcomes of the patients were classified into: a stable disease (ten patients); subsequent leukemic transformation (eight patients) and nonleukemic death (seven patients). The growth of the marrow granulocyte-macrophage progenitors (CFU-GM) at the time of diagnosis was significantly related to the survival. In addition, sequential changes in the CFU-GM growth patterns correlated with the different clinical outcome of myelodysplastic syndromes patients.

Aged↗

Detection of granulocyte colony-stimulating factor produced by a newly established human hepatoma cell line using a simple bioassay system.

A colony-stimulating factor(CSF)-producing tumor cell line (KX-87) was established from a patient with hepatocellular carcinoma and marked granulocytosis. This cell line formed tumors on nude mice in high frequency and the mice revealed marked granulocytosis. In clonogenic assays of human bone marrow cells, KX-87 conditioned medium (CM) supported the formation of colonies mainly consisting of neutrophilic granulocytes but had no burst-promoting activity. The molecular weight of the colony-stimulating activity (CSA) in KX-87CM was estimated about 25,000 daltons by gel filtration and a new bioassay system. In principle, a subline of murine hemopoietic cell line NFS-60 was cloned which was dependent on KX-87CM. Then the growth of this subline was examined by a rapid and sensitive colorimetric tetrazolium assay. From these results, it was concluded that the CSA which KX-87 cell line produced was G-CSF.

Animals↗

Effects of recombinant G-CSF and GM-CSF on in vitro differentiation of the blast cells of RAEB and RAEB-T.

To evaluate the effects of recombinant G-CSF and GM-CSF on RAEB and RAEB-T cells, blast cells from 6 patients were incubated in liquid culture systems with these CSFs for 7 days, and their numerical, morphological and functional changes were assessed. Both CSFs stimulated cell growth, but decreased the proportion of blast cells in 5 of the 6 cases. Karyotypic abnormalities persisted during cultivation in some cases. The CSFs also stimulated the expression of part of the esterase activities, and a positive interaction of both CSFs was seen in part. Although CSFs had no significant effects on the ability of cells to reduce NBT or to phagocytize latex particles, the results indicated that they induce partial differentiation of blast cells. It appears that such pathological cells still retain the capacity to respond to growth factors.

Adult↗

Nerve regeneration in the dorsal funiculus of the rat spinal cord: a light and electron microscopic study.

In an attempt to identify regenerating axons in the central nervous system, a partial transection of the dorsal funiculus in the rat spinal cord was carried out with a pair of microdissection scissors, and a nylon thread loop was inserted into the lesion to demarcate the severed tissue. Nerve regeneration through the demarcated lesion was observed 4-20 days after the operation by light and electron microscopy. In the early stage, many naked axons appeared from the caudal part of the lesion, and some of these further extended into the demarcated space. They contained an accumulation of mitochondria, smooth-surfaced endoplasmic reticulum and vesicles in the axoplasm; this axoplasmic feature indicated that they were regenerating axons. They gradually increased in number, and took highly irregular courses exhibiting various fluctuations in diameter throughout their lengths. Immature Schwann cells as well as glial cells including oligodendrocytes and astrocytes appeared in close association with these regenerating axons. Oligodendrocytes eventually formed thin myelin sheaths. On the other hand, naked axons were present deflecting outside the thread loop; they showed no axoplasmic characteristics as described above. These axons could be regarded as uninjured ones merely undergoing demyelination due to the surgery. Thus, regenerating axons were clearly distinguished from merely demyelinated ones, and some of them were shown to grow through the traumatic lesion in the dorsal funiculus of the rat spinal cord.

Animals↗

Reversal of alcohol-induced amnesia by the benzodiazepine inverse agonist Ro 15-4513.

We investigated the effects of benzodiazepine inverse agonists on ethanol-induced amnesia using a passive avoidance task. Pretraining treatment of mice with ethanol significantly impaired the passive avoidance response: there was a significant reduction in the % retention and step-down latency. The benzodiazepine inverse agonists, Ro 15-4513 and beta-CCM, significantly increased the % retention and prolonged the step-down latencies in mice treated with ethanol, but FG 7142 did not. The anti-amnesic effects of Ro 15-4513 were completely antagonized by co-administration of Ro 15-1788, a benzodiazepine antagonist. These results suggest that the anti-amnesic effect of Ro 15-4513 on alcohol-induced amnesia is mediated by benzodiazepine receptors.

Amnesia↗

Detection of carcinogenic glutamic acid pyrolysis products in Worcestershire sauce by high-performance liquid chromatography.

Commercially available Worcestershire sauce was analyzed for mutagenic and carcinogenic glutamic acid pyrolysis products using high-performance liquid chromatography. These carcinogenic heterocyclic amines were found to be present in all brands of Worcestershire sauce analyzed. The identity of the carcinogens was confirmed by spectrometric analyses and the SOS umu-test. The concentrations of 2-amino-6-methyldipyrido[1,2-a:3',2'-d]imidazole (Glu-P-1) and 2-amino-dipyrido[1,2-a:3',2'-d]imidazole (Glu-P-2) in the Worcestershire sauce were 695 +/- 329 pmol/liter (mean +/- SD, n = 5) and 1,839 +/- 1,321 pmol/liter (n = 5), respectively.

Amines↗