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Biomedical subjects

H Kimura

Publications and source records attributed to H Kimura.

At least 19 recordsLinked to original sources

Butyrylcholinesterase-rich neurons in rat brain demonstrated by a sensitive histochemical method.

Butyrylcholinesterase (BChE) is a highly active enzyme in brain, but little is known about its physiological functions. One obstacle has been the lack of a sensitive and specific method for determining its cellular localization. We report here on a histochemical technique that has permitted BChE to be detected in neuronal, glial, and vascular structures. The method, which utilizes butyrylthiocholine iodide as the substrate, is a modification of our previously described method for acetylcholinesterase (AChE) histochemistry. BChE-rich neuronal somata stained much more intensely than capillaries or glia. Prominent neuronal groups were located in the anterodorsal, laterodorsal, anteroventral, reuniens, centrolateral, paratenial, and periventricular thalamic nuclei, the laterodorsal tegmental nucleus, the pedunculopontine tegmental nucleus, and the dorsal motor nucleus of vagus. Several other areas of the forebrain and brainstem showed modest numbers of positive cells. No positive cells were detected in the striatum, hippocampus, and most parts of the hypothalamus, which are regions containing numerous AChE-rich neurons. Although the distribution pattern of BChE-rich neurons differed from that of AChE-rich neurons, some neuronal groups contained both esterases. The results suggest that BChE may play a unique role in neuronal function, particularly since many BChE-rich neurons have not been identified as to neurotransmitter type.

Acetylcholinesterase

Gamma-glutamyltaurine has potent and long-lasting antiepileptic action as demonstrated by intra-amygdaloid injection in amygdala-kindled rats.

Intra-amygdaloid injections of gamma-glutamyltaurine, a recently identified brain dipeptide, strongly suppressed seizures in amygdala-kindled rats stimulated with intensities 10 or 50 microA above the generalized seizure triggering threshold (mean 82 microA). The suppressive effect persisted as long as 3 days. Taurine had relatively weak suppressive effects. Thus gamma-glutamyltaurine seems much more potent than taurine in the suppression of epileptic seizures when injected directly into the kindling site.

Amygdala

High molecular weight basic fibroblast growth factor-like protein is localized to a subpopulation of mesencephalic dopaminergic neurons in the rat brain.

A rabbit antiserum (R917) was raised to a purified fraction of bovine brain basic fibroblast growth factor (bFGF). On Western blots of rat midbrain extract, the antiserum did not recognize low molecular weight forms of bFGF. Instead, it recognized a single band of 27-28 kDa. Immunohistochemically, the antiserum preferentially stained a subpopulation of calbindin-negative mesencephalic dopaminergic neurons. The positive somata were mainly packed in a ventral portion of the tegmentum including the A10 region, the ventral tegmental area and the pars compacta of the medial substantia nigra, but were also scattered in both the pars compacta and reticulata portions of the lateral substantia nigra. Processes of dendrites and axons were clearly visible. Terminal fields were located in striosomes, the dorsolateral rim of the neostriatum, the anterodorsal aspect of the nucleus accumbens shell, the infralimbic cortex, and the medial prefrontal cortex. These results suggest that trophic specialization in subpopulations may occur in all three of these dopaminergic projection systems, i.e. the nigrostriatal, mesolimbic and mesocortical pathways.

Animals

Effect of carbachol on luminal release of somatostatin from isolated perfused rat duodenum.

The dynamic release of somatostatin-like immunoreactivity (SLI) from duodenum into the lumen was studied in the isolated, vascularly perfused rat duodenum. The luminal release of duodenal SLI was stimulated by a cholinergic agonist, carbachol, and the carbachol-induced release of SLI was completely blocked by atropine, but not by hexamethonium. These data suggest that the luminal release of SLI from rat duodenum is under the control of a cholinergic muscarinic stimulation. The ratio of somatostatin-14 to somatostatin-28 in picograms was about 1 during basal release but increased to approximately 2 during carbachol stimulation.

Animals

Behavior of VRG neurons during the atonia of REM sleep induced by pontine carbachol in decerebrate cats.

The microinjection of carbachol into the pons of acute decerebrate cats elicits a REM sleep-like atonia and a profound suppression of respiratory motoneuronal activity (J. Appl. Physiol., 69 (1990) 2280-2289). To assess whether this suppression is mediated by medullary neurons that provide respiratory drive to motoneurons of the respiratory pump muscles (diaphragm and intercostals), we studied the effect of pontine carbachol on the activity of neurons of the ventral respiratory group (VRG) in decerebrate, vagotomized, paralyzed and artificially ventilated cats. VRG neurons were recorded extracellularly along with the activity of phrenic and intercostal (external and internal) nerves. Both inspiratory (I) and expiratory (E) VRG neurons had incrementing, ramp-like bursts of activity during their firing periods and were not vagal motoneurons. Carbachol produced a depression of the peak firing rate in most (42/57) neurons studied. However, five cells showed no change and ten had an increase in activity in spite of consistent depression at the motoneuronal level. For the total population of cells (34 I and 23 E), the peak firing was reduced to 88.5% +/- 16.3 (S.D.) of control. The simultaneously recorded phrenic activity was reduced to 77.9% +/- 11.5, while inspiratory intercostal activity fell to 63.4% +/- 21.6 and expiratory to 23.2% +/- 21.2 of control. The carbachol-induced changes in peak firing of both I and E cells were quantitatively similar, and positively correlated to changes in peak phrenic activity. Analysis of this correlation suggested that phrenic and intercostal activities will be depressed to some degree by carbachol even when the average VRG cell activity remains unchanged. In addition, our data show that VRG cells may receive a combination of inhibitory and excitatory inputs during the carbachol-induced depression of respiratory motoneurons. Thus, although some disfacilitation from VRG cells may occur, there must be additional inhibitory or disfacilitatory pathways that mediate the decrease in activity of both phrenic and intercostal motoneurons that accompanies the REM sleep-like atonia.

Animals

Growth factors and vitamin E modify neuronal glutamate toxicity.

The sympathetic nerve cell line PC-12 is killed by glutamate in a concentration-dependent manner. Although glycine and the deletion of magnesium weakly potentiate glutamate toxicity and PC-12 cells express N-methyl-D-aspartate-receptor mRNA, most toxicity is mediated by means of a mechanism independent of typical N-methyl-D-aspartate receptors. Glutamate toxicity is, however, greatly enhanced by prior exposure to nerve growth factor or basic fibroblast growth factor. Glutamate killing is blocked by epidermal growth factor and, to a lesser extent, by vitamin E. These observations show that synergistic interactions between growth factors and excitotoxic amino acids may play critical roles in the developing nervous system and that antioxidants attenuate this toxicity.

Animals

Monoclonal antibody to GABA binding protein, a possible GABAB receptor.

A monoclonal antibody has been raised against a partially purified preparation for the GABAB receptor. The antibody recognized a protein of about 80 kDa in bovine brain synaptic membrane. Immunoabsorbent agarose beads conjugated with the antibody were able to remove, without visible changes in electrophoresed profiles of total proteins, over 90% of the baclofen suppressive GABA binding activity (designated herein, GABAB receptor binding activity) in the solubilized synaptic membrane fraction. Moreover, the addition of GB-1 antibody directly inhibited the GABA binding activity in the crude synaptic membrane fraction. These results indicate that the monoclonal antibody obtained here recognizes the GABA binding protein, or more specifically a GABAB receptor.

Animals

Developmental changes in the serotoninergic innervation of hindlimb extensor motoneurons in neonatal rats.

The postnatal development of quadriceps femoris motoneurons (Q-MNs) and serotonin (5-HT) nerve terminals in rat spinal cord were studied using retrograde neurotracing techniques combined with 5-HT immunohistochemistry. We attempted to elucidate the 5-HT-ergic innervation to the Q-MNs by counting the number of 5-HT-immunoreactive varicosities that were in close apposition to the Q-MNs. The following results were obtained: (1) Q-MNs possessed, at birth, few if any very short dendrites. The size of these somata was relatively uniform and small. During postnatal periods lasting from 1 to 30 days, the mean cell size of Q-MNs increased with the development of dendrites. From 5 to 14 days after birth, in particular, cell size increased markedly. (2) 5-HT-immunopositive fibers were, at birth, already observed in the ventral horn of the lumbar spinal cord. The density of these fibers increased gradually with age. (3) At birth, only a few 5-HT terminals and varicosities showed close apposition with about half the Q-MNs examined. At 5-days postnatally, such close apposition was found in all Q-MNs. By the first two postnatal weeks, Q-MNs grew quickly and the 5-HT innervation to the Q-MNs appeared to have been established. Based on these results, the significance of 5-HT innervation to developing Q-MNs is discussed in relation to the postnatal development of motor function.

Animals

Isolation and characterization of macrophages from rat embryonic muscle culture.

We have previously described the wide distribution of resident macrophages in normal rat skeletal muscle. In this study, we investigated the characteristics of the macrophages that occur in rat embryo muscle cultures. We showed that cells of monocyte-macrophage lineage are present in primary muscle cultures of rat embryos (18 days in gestation) and that these cells form morphologically and phenotypically heterogeneous populations, based on their reaction with monoclonal antibodies ED1, ED2, ED3, and OX43. Constitutively Ia+ cells with dendritic appearance were also observed. Furthermore, we established the procedure for isolation of macrophages from the primary muscle cultures. The isolated cells, mostly ED1+, expressed class I and CD4 antigens and bore complement (C3) receptors on their surfaces. The fact that cell of monocyte-macrophage lineage occur in the embryonic muscle suggests that during embryogenesis these cells may enter the developing muscle and give rise to a population of tissue-associated macrophages.

Animals

Proliferative activity in gastric cancer determined with cell cycle-related monoclonal antibodies Ki-67 and p105: analysis by flow cytometry.

The proliferative activity of gastric cancer cells was determined by DNA flow cytometric (FCM) analysis and labeling rates of Ki-67 and monoclonal antibodies and proliferation-associated nuclear antigen (p105) autoantibodies in 28 patients with fresh human gastric cancer cells. By setting the cutoff line at the level as used in a negative control study without primary antibody in the same sample, the Ki-67 and p105 labeling rates were calculated by the dual fluorescence analysis. A total of 43 experiments was performed on FCM analysis for each antigen: 28 with Ki-67 and 15 with p105. The mean Ki-67 labeling rate of gastric cancer cells was 45.1% (13.9-76.3%). The Ki-67 labeling rates were significantly higher for larger size tumor, peritoneal metastasis, and advanced clinical stage. A significant correlation was found between Ki-67 labeling rate and p105 labeling rate (P < 0.05). Bivariate FCM may be an easy method for obtaining useful information of cell kinetics.

Antibodies, Monoclonal

Independent clinical and flow cytometric prognostic factors for the survival of patients with stage I gastric cancer.

Paraffin-embedded tumor samples from 151 patients with stage I gastric cancer were analyzed by DNA flow cytometry, and 80 patients received an infusion of bromodeoxyuridine (BrdU) to determine S-phase fraction. S-phase fractions of tumors were measured by the immunohistochemical method using anti-BrdU monoclonal antibody. Of the 151 patients, 81 (54%), and 70 (46%) showed diploid and aneuploid patterns. There was no significant association between DNA ploidy and wall invasion, histologic type, or lymphatic invasion. Aneuploid tumors were associated with positive-vessel invasion. When the DNA ploidy and clinicopathological parameters were simultaneously entered into the Cox regression model, DNA ploidy and wall invasion emerged as independent prognostic parameters. Aneuploid tumors had significantly higher values of BrdU labeling indices than diploid ones. These results indicate that the determination of DNA ploidy patterns may be an important prognostic factor in patients with stage I gastric cancer, and may be useful in deciding the therapeutic schedule of patients with gastric cancer.

Aneuploidy

The succinate dehydrogenase inhibition test for evaluating biopsy specimens and resected tumors of advanced gastric cancer.

An in vitro chemosensitivity study of both biopsy specimens and surgically resected tumors of advanced gastric cancer from 12 patients was evaluated using the succinate dehydrogenase inhibition (SDI) test. A decrease in succinate dehydrogenase (SD) activity as an indicator of chemosensitivity was determined using cisplatin (CDDP), etoposide (VP-16), and mitomycin C (MMC). In this study, 29 of a total 36 experiments were evaluable (80.6%) and significant correlations were found in all three of the antitumor drugs (P < 0.03). This finding suggests that the SDI test using biopsy specimens may prove valuable for assessing the preoperative chemosensitivity of advanced gastric cancer.

Aged

Recurrence of superficial bladder tumours after transurethral resection.

From 1979 to January 1990, 48 patients with newly diagnosed superficial bladder tumours were treated by transurethral resection (TUR) at Toyama Medical and Pharmaceutical University Hospital. The relationship between tumour recurrence and the stage, grade, number, or size of the tumours was investigated. The 1-, 2-, 3-, 4- and 5-year non-recurrence rates were 72%, 63%, 47%, 40%, and 40%, respectively. Non-recurrence rate for pTa tumours was significantly higher than that for pT1 tumours (p < 0.05). There was no relationship between tumour grade and recurrence. The rate of recurrence of multiple tumours was higher than that of single ones. There was no difference in non-recurrence rate between tumours smaller than 1 cm and those of 1 cm or larger. Non-recurrence rate in the instillation therapy group was significantly higher than in the non-instillation group (p < 0.05). It was concluded that patients with multiple or high-stage tumours have the risk of a high rate of recurrence and that intravesical chemotherapy is effective in preventing local recurrence in some patients.

Administration, Intravesical

Expression of a human multidrug resistance gene in human ovarian carcinoma cell lines.

To investigate the possible role of the multidrug resistance phenotype to chemoresistance in human ovarian carcinoma, we have analyzed human multidrug resistance gene (mdr 1) expression in 8 human ovarian adenocarcinoma cell lines. An increase in P-glycoprotein level specific to multidrug-resistant tumor cells was not apparently associated with the increase in resistance to vincristine (VCR) or doxorubicin (Adriamycin). Mdr 1 transcripts (4.5 kilobases) were observed in the RNA preparation obtained from only one cell line (SHIN-3) that showed the highest resistance to both drugs in vitro and in vivo. No cell lines showed mdr 1 DNA amplification. These results suggest that the insensitivity of human ovarian carcinoma to chemotherapy could be partly explained by the expression of mdr 1.

ATP Binding Cassette Transporter, Subfamily B, Mem

Intraoperative endoscopic electrohydraulic lithotripsy of pancreatic stones.

Two male patients with complications associated with chronic pancreatitis are described. In each patient, preoperative examinations revealed a large stone obstructing the main duct in the head of the pancreas. Lateral pancreaticojejunostomy was performed to relieve pain and prevent further attacks of pancreatitis. During each operation, the stone was fragmented under direct visual control with the use of a flexible choledochoscope and a contact electrohydraulic lithotriptor. The stone was removed and ductal flow through the head of the pancreas was reestablished. Our experience shows that endoscopic electrohydraulic lithotripsy facilitates operative removal of pancreatic stones deeply located in the head of the pancreas.

Adult

Multiple endocrine neoplasia type 2 with malignant pheochromocytoma--long term follow-up of a case by 131I-meta-iodobenzylguanidine scintigraphy.

The case of a 33-year-old Japanese man, who has Multiple Endocrine Neoplasia Type 2 (MEN IIa) (Sipple's syndrome) with malignant pheochromocytoma, is reported. He had survived for twelve years since the initial diagnosis of malignant pheochromocytoma. Within this period, he had undergone 131I-meta-iodobenzylguanidine scintigraphy twice, in 1983 and 1990. This is the first case in Japan of a longterm surviving patient with malignant pheochromocytoma followed up by 131I-MIBG scintigraphy. Although he had no exacerbation of clinical symptoms or urinary catecholamine levels, second scintigraphy clearly showed an increase in the tumor size, new metastasis of the malignant pheochromocytoma and exacerbation of the medullary thyroid carcinoma. Compared with any other roentgenological device and hormonal data, 131I-MIBG scintigraphy was seen to be a good tool for evaluating the localization and the progression of tumors. 131I-MIBG scintigraphy is a useful procedure not only for initial diagnosis but also for judging progression in a case of advanced malignant pheochromocytoma.

3-Iodobenzylguanidine

Impairment of the TSH signal transduction system in human thyroid carcinoma cells.

In order to further evaluate the role of TSH in the proliferation and the differentiation of human thyroid carcinoma cells, we have analyzed the function of the TSH receptor in the established thyroid carcinoma cell lines NPA and WRO. The TSH signal transduction system in the carcinoma cells was also compared with that in normal thyroid cells. Although unresponsiveness to bovine and human TSH was demonstrated by measurement of cAMP production and [3H]thymidine incorporation after treatment of TSH, cAMP production was induced after stimulation of these cells by forskolin, cholera toxin, and isoproterenol. Specific binding to 125I-TSH was demonstrated in both NPA and WRO cells in addition to the existence of a TSH receptor mRNA and thyroglobulin mRNA species, although thyroid-specific gene expression in these cells was not regulated by TSH. These findings suggest that the unresponsiveness to TSH in these cells may be due to an abnormality of TSH receptor-G protein coupling rather than to a decreased level of TSH-receptor expression or a Gs protein abnormality.

Base Sequence