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Biomedical subjects

Y Matsumoto

Publications and source records attributed to Y Matsumoto.

At least 19 recordsLinked to original sources

The p53 tumor-suppressor gene and ras oncogene mutations in oral squamous-cell carcinoma.

The frequencies of mutations in the p53 tumor-suppressor gene and ras proto-oncogenes were investigated systematically in surgically resected oral squamous-cell carcinomas (SCCs) using single-strand conformation polymorphism (SSCP) and/or dot-blot hybridization analysis of DNA fragments which had been amplified by the polymerase chain reaction (PCR). p53 gene mutations, within the region of exons 5 to 8, were detected in 17 out of 27 (63%) tumor specimens. The role of p53 mutations in cell-line establishment was investigated. p53 gene mutations were detected in 5 out of 6 tissue samples from which cell lines were established and in 4 out of 5 specimens from which cell lines could not be established, suggesting that the presence of p53 gene mutations is not by itself sufficient for cell-line establishment. Tumor samples were also analyzed for point mutational activation of the ras proto-oncogenes. One out of 30 (3%) tumors showed an activating point mutation in codon 12 of H-ras, this being consistent with reports from Europe and USA but not with any from India. Compared to frequencies of the other genetic changes so far reported for oral SCC, the p53 mutations have been observed most often to undergo genetic change. p53 gene mutation is thus intimately involved in the genesis of oral SCC and consequently should be useful as a marker for the diagnosis of this neoplasm.

Adult

Development of peptide- and tyrosine hydroxylase-containing neurons in the fetal spinal cord transplanted into the anterior chamber of the eye of adult rats.

Fetal rat spinal cord transplanted into the anterior chamber of the eye of an adult rat was immunohistochemically stained using antisera to substance P (SP), neuropeptide Y (NPY), methionine-enkephalin (ENK), vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP) and tyrosine hydroxylase (TH), and distributional changes of peptide- and enzyme-containing neurons 1, 2 and 4 weeks after transplantation were investigated. To examine the effect of colchicine on immunoreactivity, unilateral eyes of these adult host rats received intraocular colchicine treatment. Without colchicine treatment, numerous SP- and CGRP-immunoreactive (IR) neurons were observed in the graft 1 week after transplantation, and their immunoreactivity gradually decreased up to 4 weeks after transplantation. NPY-, ENK-and VIP-IR neurons first appeared in the graft 2 weeks after transplantation. Four weeks after transplantation, the immunoreactivity of NPY and ENK decreased significantly, whereas VIP-IR neurons showed the same intensity as that observed at 2 weeks after transplantation. TH-IR neurons, on the other hand, were seen at every stage, but their immunoreactivity was constant all the time. After colchicine treatment, the number of SP-, NPY-, ENK- and CGRP-IR neurons appeared to increase, while that of VIP- and TH-IR neurons did not change significantly. The distribution patterns of the peptide- and enzyme-containing fibers differed from each other. In the analysis of serial sections stained with 5 peptides (SP, NPY, ENK, VIP, CGRP), fibers containing these peptides were found to be densely accumulated in specific areas of the transplanted spinal cord. The present findings demonstrated that most of the peptide- and enzyme-containing neuron systems in the transplanted spinal cord showed similar distribution patterns and development to those in the normal spinal cord, but that some displayed different distribution.

Animals

Relationship between cytosolic Ca2+ concentration and amylase release in rat parotid acinar cells following muscarinic stimulation.

Carbachol (CCh), a muscarinic-cholinergic agonist, increased both cytosolic free calcium concentration ([Ca2+]i) and amylase release in rat parotid acinar cells or acini in a dose-dependent manner. Treatment of acinar cells with the intracellular Ca2+ antagonist, 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8), or the intracellular Ca2+ chelator, 1,2-bis(O-aminophenoxy)ethane-N,N,N'N'-tetraacetic acid (BAPTA), strongly attenuated the increases in [Ca2+]i evoked by CCh, but amylase release from acini was not significantly suppressed by the treatment with TMB-8 or BAPTA. Low concentrations (0.02-0.5 microM) of ionomycin, a Ca2+ ionophore, caused increases in [Ca2+]i comparable to those induced by CCh, but the same concentrations had only a little effect on amylase release. The protein kinase C activator, 12-O-tetradecanoylphorbol-13-acetate (TPA), stimulated amylase release in quantities similar to those induced by CCh, although TPA alone did not cause any change in [Ca2+]i. Combined addition of TPA and ionomycin potentiated only modestly amylase release stimulated by TPA alone. Staurosporine, a protein kinase C-inhibitor, similarly inhibited both the CCh- and TPA-induced amylase release. These results suggest that an increase in [Ca2+]i elicited by CCh does not play an essential role for inducing amylase release in rat parotid acini. Amylase release by muscarinic stimulation may be mediated mainly by activation of protein kinase C.

Alkaloids

Effects of Y-20811, a thromboxane A2 synthetase inhibitor, on experimentally induced coronary thrombosis in anesthetized dogs.

The effects of Y-20811, a selective inhibitor of thromboxane A2 (TXA2) synthetase, on blood flow and local levels of immunoreactive thromboxane B2 (i-TXB2) and 6-keto prostaglandin F1 alpha (i-6-keto PGF1 alpha) in the coronary artery were investigated in the canine model of coronary thrombosis. Thrombosis was induced by applying an electric current to the intraluminal surface of the coronary artery. The plasma levels of i-TXB2 and i-6-keto PGF1 alpha were measured distal to the electrode. In the control group, coronary blood flow decreased and finally stopped 207 +/- 53 min (mean +/- S.E.M., n = 5) after the start of current application. The level of i-TXB2 rose before the coronary occlusion. Coronary blood flow did not change significantly in the Y-20811-treated group (1 mg/kg i.v.). The level of i-TXB2 decreased and remained significantly lower than that in the control group. The level of i-6-keto PGF1 alpha tended to increase slightly in the Y-20811-treated, but not in the control group. The weight of the thrombus in the Y-20811-treated group was significantly less than that in the control group (P less than 0.01). These results suggest that Y-20811 prevents coronary thrombosis by the inhibition of TXA2 production around the electrically injured lumen of the coronary artery.

6-Ketoprostaglandin F1 alpha

Altered zonal expression of the CD26 antigen (dipeptidyl peptidase IV) in human cirrhotic liver.

Dipeptidyl peptidase IV is a cell surface ectopeptidase with widespread tissue distribution. Recently it was shown to display extracellular matrix-binding properties; therefore its role in cirrhosis is of interest. The aim of this study was to use monoclonal antibodies directed against the human CD26 antigen (which has been shown to be dipeptidyl peptidase IV) to study the distribution of this molecule in normal human and cirrhotic liver. Identical staining was obtained with the three monoclonal antibodies (TaI, 1F7 and TS145) and enzyme histochemistry. In normal liver (n = 11) intense staining of hepatic acinar zones 2 and 3 was present, but little staining was seen in zone I. Hepatocyte staining was confined to the bile canalicular domain. In cirrhotic livers (n = 23) obtained at transplantation, staining of regenerating nodules without a zonal pattern was present. In addition, we saw staining of the lymphoid cell infiltrate and proliferating bile ductules. In a minority of cirrhotic biopsy specimens (four) staining of the basolateral hepatocyte domain in regenerating nodules was seen. Biopsy specimens from hepatic allografts (n = 28) were used as disease controls. These samples all showed preferential staining of zones 2 and 3, similar to that in normal biopsy specimens. Eleven of these samples showed staining of the basolateral and bile canalicular domains. In conclusion, the normal acinar distribution of dipeptidyl peptidase IV (zones 2 and 3) is lost in cirrhotic nodules. Furthermore, the altered membrane distribution of this molecule in cirrhosis and allograft rejection may allow increased hepatocyte extracellular matrix interactions during organ remodeling.

Antibodies, Monoclonal

Transport of drugs through human erythrocyte membranes: pH dependence of drug transport through labeled human erythrocytes in the presence of band 3 protein inhibitor.

To reveal the role of the band 3 anion transport protein of the erythrocyte membrane in drug transport through the membrane, the possible effects of inhibitors of anion transport on the permeability of some anionic drugs were examined. The amounts of these drugs that permeated varied markedly with the pH of the outer medium around human erythrocytes that contained the band 3 protein inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS). In the pH range 7.0-8.0, the membrane permeability of drugs through DIDS-treated erythrocytes was affected by a slight pH change (0.2), whereas that through the intact erythrocytes was not pH dependent. These results suggest that the band 3 protein acts not only as a channel for the transport of anions or some anionic drugs but also as protection for the transport system from changes in the pH of the outer medium.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid

Fluorescent dye rhodamine 6G as a molecular probe to study drug resistance of C6 rat glioma cells.

A study was made of the membrane transport of cytoplasm and mitochondria stained fluorescence dye Rhodamine 6G (R6G). In rat glioma C6 cells and 1-(4-amino-2-methyl-5-pyrymidinyl)-methyl-3-(2-chloroethyl) -3-nitrosourea hydrochloride (ACNU) and vincristine (VCR) resistant cell lines (C6/ACNU, C6/VCR), the rate of uptake of R6G decreased in C6/VCR cells, but verapamil increased the intracellular accumulation of R6G in C6/VCR. The intracellular accumulation of R6G of C6/ACNU cells was essentially the same as that of wild-type cells. C6/ACNU cells did not show cross resistance and were sensitive to VCR and cisplatin. C6/VCR cells showed cross resistance to ACNU and CDDP, but C6/VCR cells in the presence of verapamil were more sensitive to drugs than C6/VCR cells in the absence of verapamil. We conclude that the reduction of R6G fluorescence staining intensity in C6/VCR cells compared to wild-type cells may be associated with the mechanism of multidrug resistance (MDR) but does not reflect the mechanism of resistance to ACNU. Verapamil increased the accumulation of R6G in C6/VCR cells and overcame MDR, suggesting that there is a correlation between the MDR overcoming effect and enhancement of R6G accumulation, and that this correlation validates the use of the R6G staining test for clinical and laboratory investigation of MDR.

Animals

Anticentromere-protein-B--DNA complex activities in anticentromere antibody-positive patients.

Centromere protein B (CENP-B), which is an alphoid DNA binding protein, is the target antigen in autoimmune disease patients (often those with scleroderma). In this study, we analysed activities of anti-CENP-B-DNA complex in anticentromere antibody (ACA)-positive patients using DNA immunoprecipitation with purified CENP-B. The activities correlated with ACA titres and were closely associated with Raynaud's phenomenon. Patients with CREST symptoms (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, telangiectasia) showed higher activities than those with no symptoms. Our results suggest that autoimmune responses to native CENP-B may have an important role in the pathogenesis of scleroderma.

Autoantibodies

Day-to-day changes in oxygen uptake kinetics at the onset of exercise during strenuous endurance training.

The aim of this study was to assess the effect of strenuous endurance training on day-to-day changes in oxygen uptake (VO2) on-kinetics (time constant) at the onset of exercise. Four healthy men participated in strenuous training for 30 min.day-1, 6 days.week-1 for 3 weeks. The VO2 was measured breath-by-breath every day except Sunday at exercise intensities corresponding to the lactate threshold (LT) and the onset of blood lactate accumulation (OBLA) which were obtained before training. Furthermore, an incremental exercise test was performed to determine LT, OBLA and maximal oxygen uptake (VO2max) before and after the training period and every weekend. The 30-min heavy endurance training was performed on a cycle ergometer 5 days.week-1 for 3 weeks. Another six men served as the control group. After training, significant reductions of the VO2 time constant for exercise at the pretraining LT exercise intensity (P less than 0.05) and at OBLA exercise intensity (P less than 0.01) were observed, whereas the VO2 time constants in the control group did not change significantly. A high correlation between the decrease in the VO2 time constant and training day was observed in exercise at the pretraining LT exercise intensity (r = -0.76; P less than 0.001) as well as in the OBLA exercise intensity (r = -0.91; P less than 0.001). A significant reduction in the blood lactate concentration during submaximal exercise and in the heart rate on-kinetics was observed in the training group.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

An immunoserological study of patients with vibration syndrome.

In a study undertaken to investigate the relationship between vibration syndrome and collagen disease, 90 Japanese patients with vibration syndrome (all men aged 41-86 years) were examined immunologic abnormalities, mainly by a set of immunoserology tests. Of these 90 patients, 25 had examinations at yearly intervals for 3 consecutive years, while the other 65 underwent examinations once. The results indicate that, of all the patients studied, 10 (11.1%) gave a positive RA test, 6 (6.7%) had leukopenia, 5 (5.6%) had hypergammaglobulinemia, and 15 (16.7%) had hypocomplementemia (CH50). Worthy of particular note are the 21 patients (23.3%) who were positive for nuclear-specific antibodies (1: greater than or equal to 40) (using Hep-2 cells as the nuclear substrate), with some of them suspected of having Sjögren's syndrome, progressive systemic sclerosis, or systemic lupus erythematosus. During a 3-year follow-up period, 10 (40%) of 25 patients exhibited rising titers of nuclear-specific antibodies with conversion to seropositivity for these antibodies. The facts that the positivity rate for nuclear-specific antibodies was significantly higher in these patients with vibration syndrome (23.3%) than in healthy adult men over 40 years of age (1.8%) (P less than 0.05) and that a progressive elevation of nuclear-specific antibody titer was noted in a high percentage of the patients were suggestive of some causal relationship between the appearance of nuclear-specific antibodies and the use of vibrating tools.

Adult

Circulating myosin light chain I levels after coronary reperfusion: a comparison with myocardial necrosis evaluated from single photon emission computed tomography with pyrophosphate.

This study was performed to assess the influence of coronary reperfusion on the serial serum myosin light chain (LC)I levels and to evaluate the relationship between the peak LCI level and the infarct size calculated from single photon emission computed tomography (SPECT) with technetium-99m pyrophosphate (Tc-99m PYP) in 11 patients who underwent coronary reperfusion. Blood was drawn before reperfusion, immediately after reperfusion, and once a day for 14 days, to estimate the time course of serum LCI release. The infarct size estimated by Tc-99m PYP ranged from 7.3 to 62.4 ml. The LCI levels obtained before reperfusion were less than 2.5 ng/ml but those obtained immediately after reperfusion were much higher. The value ranged from 2.7 to 9.7 ng/ml and that expressed as a percentage of peak LCI (% peak LCI) ranged from 19 to 83%. Collateral circulation, reperfusion arrhythmia and the degree of residual stenosis had no influence upon the % peak LCI. The correlation between peak LCI levels and SPECT-determined infarct size was good, with a correlation of 0.76 (p less than 0.01, regression line by least squares method y = 3.31 + 1.53x). Early serum LCI might be influenced by coronary reperfusion but the peak LCI value reflected acute myocardial necrosis in patients who underwent coronary reperfusion.

Adult

Surgical treatment of primary carcinoma of the gallbladder based on the histologic analysis of 48 surgical specimens.

During the past 5 years, extended cholecystectomy and/or more radical procedures, including pancreatoduodenectomy and extended hepatic lobectomy, were performed on 48 patients with carcinoma of the gallbladder in an attempt to cure the disease. It was retrospectively proven from microscopic analysis of the surgical specimens taken from the 48 patients that 28 patients had received curative resections. The 28 consisted of all 4 patients with stage I carcinoma, all 9 with stage II, 6 of 8 with stage III, and 9 of 27 with stage IV. We reached the following conclusions: (1) extended cholecystectomy with resection of the bile duct in the hepatoduodenal ligament is the optimum procedure for patients with stage I and stage II carcinomas, and (2) for patients with stage III carcinoma, more radical procedures, including pancreatoduodenectomy and/or wider resection of the liver, might result in curative resection of the disease. For some patients with stage IV tumors, such as duodenal invasion or direct invasion of the liver, curative resection may be achieved by pancreatoduodenectomy or extended hepatic lobectomy.

Adult

Detection of enhancer repeats in the long terminal repeats of feline leukemia viruses from cats with spontaneous neoplastic and nonneoplastic diseases.

Enhancer duplication in the long terminal repeat of feline leukemia virus (FeLV) was examined in primary cells from naturally FeLV-infected cats with various neoplastic and nonneoplastic diseases using the polymerase chain reaction. In all cases, a 170-bp band, corresponding to a standard exogenous FeLV with one copy of enhancer, was detected. Repeated enhancer sequences were found in all 8 cases of thymic-form lymphosarcoma, in some cases of lymphosarcoma of other forms (3/8) and myeloid tumors (2/3), and in only 1 of 6 cases with nonneoplastic diseases. The copy number of FeLV proviruses with a repeated enhancer seemed higher than that of those with one copy of enhancer in 3 cases of thymic form lymphosarcoma. In 5 cases of thymic form lymphosarcoma and in 1 case of erythroleukemia, coexistent FeLVs with double and triple enhancers of different sizes were found. Of the enhancer elements, only the SV40 core binding site was found in all the enhancer direct repeats of these FeLVs. All the provirus clones with single and duplicated enhancer sequences from a single tumor showed mutations or deletions characteristic to that tumor, indicating that enhancer repeats may arise in individual animals after infection with a single virus clone. The present findings indicate that FeLV with enhancer repeats generated in the cat is associated with the induction of neoplastic diseases in natural conditions.

Animals

Immunohistochemical and enzymehistochemical studies of peptidergic, aminergic and cholinergic innervation of the lacrimal gland of the monkey (Macaca fuscata).

The distribution of nerve fibers containing peptides which include calcitonin gene-related peptide (CGRP), substance P (SP), neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) and enzymes of tyrosine hydroxylase (TH) and acetylcholinesterase (AchE) in the lacrimal gland of the monkey (Macaca fuscata) was studied using immunohistochemical and enzymehistochemical methods. We also examined the trigeminal ganglion (TG) and superior cervical ganglion (SCG) using the same methods. All peptide- and enzyme-containing nerve fibers examined in this study were present in the lacrimal gland and a consistent distribution pattern for each substance was found. CGRP-immunoreactive (IR) nerve fibers were mainly distributed around the blood vessels in the interlobular connective tissue. The distribution pattern of SP-IR nerve fibers was similar to that of CGRP-IR nerve fibers, but they were much less in number. NPY-IR nerve fibers were observed mostly around the blood vessels and occasionally in the interstitial stroma between the acini. Numerous VIP-IR nerve fibers were found surrounding the acini, ducts and blood vessels. TH-IR nerve fibers were also been around the blood vessels and in the interstitial stroma between the acini, as were NPY-IR fibers. The highest concentration of acetylcholinesterase (AchE)-positive nerve fibers was present in the acini, ducts and blood vessels, showing a similar distribution to VIP-IR fibers. In the TG, 50% of medium and 30% of small ganglion cells were CGRP-IR cells, while 20% of medium and 25% of small ganglion cells were of the SP-IR types.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholinesterase

Microglial and astroglial reactions to inflammatory lesions of experimental autoimmune encephalomyelitis in the rat central nervous system.

Gliosis is a repair process of lesions appearing in the central nervous system (CNS). Although gliosis by astrocytes (astrocytic gliosis) has been well documented, that by microglia (microglial gliosis) remains poorly understood. In the present study we induced experimental autoimmune encephalomyelitis (EAE) in Lewis rats and examined microglial and astroglial reactions to EAE lesions at various stages of the disease by immunohistochemistry. For the demonstration of microglia and astrocytes, antibodies against complement receptor type 3 (OX42) and glial fibrillary acidic protein (GFAP) were used, respectively. It was revealed that the whole course of microglial and astroglial reactions to EAE lesions is divisible into three stages, i.e., initial, peak and recovery stages. Microglial and astroglial reactions to EAE lesions at each stage correspond well with the clinical and histological stages of EAE. At the initial stage, rats showed mild clinical signs and a few inflammatory foci were found in the CNS. Microglia were increased in number in close association with inflammatory cell aggregates, whereas astrocytes showed no significant reaction in spite of the presence of inflammatory cells. At the peak stage, rats showed full-blown EAE and the number of inflammatory cells reached maximum. The most characteristic finding at this stage was 'encasement' of inflammatory lesions by astrocytic fibers. Microglia were increased in number, but association of microglia with lesions was prevented by astrocytes. Interestingly, however, such characteristic distribution of microglia and astrocytes was not observed at the recovery stage. Residual inflammatory cell aggregates were intermingled with dense microglial and astrocytic gliosis, forming 'micro-astroglial scars'. Double immunofluorescence staining with anti-GFAP and anti-bromodeoxyuridine (BrdU), or with OX42 and anti-BrdU revealed that BrdU-incorporated microglia, but not astrocytes, were present mainly at the initial and peak stages, suggesting that microglia would proliferate by cell division to create gliosis, whereas astrocytic gliosis would be a result of migration of astrocytes and/or up-regulation of expression of GFAP molecule. Taken together with previous in vitro findings that microglia, but not astrocytes, stimulate encephalitogenic T cell proliferation, these in vivo findings suggest that microglia augment, whereas astrocytes suppress, inflammatory processes in the CNS.

Animals

Immunocytochemical survey of haloperidol-induced immunoreactive changes of [Met]enkephalin-Arg6-Gly7-Leu8 in the rat forebrain.

It has already been demonstrated that chronic treatment with the dopamine receptor blocker, haloperidol, results in an increase of proenkephalin-A-derived peptides in the caudate-putamen (CP). To examine this phenomenon at the cellular level, we used immunocytochemistry to investigate the effects of haloperidol on [Met]enkephalin-Arg6-Gly7-Leu8 (MEAGL) immunoreactivity in the rat forebrain. After daily haloperidol (5 mg/kg, IP, for 6 days) or haloperidol decanoate (70 mg/kg, IM, given once or twice) treatment, immunoreactive neurons appeared diffusely in the whole CP and in the core part of the nucleus accumbens (Acb) and less frequently in the outer shell part of the Acb and the cell-dense layer of the tuberculum olfactorium (TuO). Increase of MEAGL-immunoreactive fibers in the CP, Acb, and TuO was also detected after these treatments, a particularly prominent increase being found in the striopallidal terminals in the globus pallidus and ventral pallidum. Haloperidol or haloperidol decanoate had no effect on MEAGL immunoreactivity in the cerebral cortex, amygdala, or hypothalamus. Reserpine treatment (5 mg/kg, IP, for 6 days) caused similar effects on the dorsal and ventral striopallidal system, and the direct injection of 6-hydroxydopamine (10 micrograms/5 microliters) into the CP led to the appearance of MEAGL-immunoreactive neurons in accordance with the depleted dopaminergic terminal area. These findings suggest that haloperidol influences enkephalinergic neurons region specifically and that in the dorsal and ventral striopallidal enkephalinergic system haloperidol increases MEAGL immunoreactivity in cell bodies, fibers, and terminals by blocking intrastriatal dopaminergic neurotransmission.

Animals