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Biomedical subjects

M Miyake

Publications and source records attributed to M Miyake.

At least 271 records · Page 15Linked to original sources

Treatment for stage III-IV neuroblastoma patients: initial response to chemotherapy evaluated by biochemical parameters.

Fourteen patients with stage III-IV neuroblastoma were treated with alternating combination chemotherapy consisting of (a) VP16/cisplatin and (b) doxorubicin/vincristine/cyclophosphamide. The initial response to induction chemotherapy, especially to VP16/cisplatin was evaluated by determining t 1/2 for urinary vanillylmandelic acid (VMA), homovanillic acid and serum neuron specific enolase (NSE). The period prior to normalization of these parameters was also determined. The patients could be classified as 7 rapid responders, with less than 3 weeks of t 1/2 VMA, or t 1/2 NSE, and 7 slow responders longer than 4 weeks of t 1/2 VMA. An analysis of the data indicates that an initial rapid response correlated with subsequent high complete response rate, but did not necessarily predict better prognosis in these patients.

Abdominal Neoplasms↗

Vitamin E and the peroxidizability of erythrocyte membranes in neonates.

We showed the increased susceptibility of neonatal biomembranes to oxidation by a kinetic analysis using an azo compound as a free-radical initiator and red blood cell (RBC) ghosts as a model membrane. When the RBC ghosts were oxidized, oxygen consumption was suppressed during the induction period in which membrane tocopherol was consumed at a constant rate, while increased oxygen uptake was observed after the tocopherol was exhausted. The total tocopherol content was similar in cord, maternal, and adult RBC ghosts, and there were no differences in the induction period (tinh) among the three types of ghosts. While the oxygen uptake rate during the induction period (Rinh) was similar in cord and adult ghosts, the rate in the subsequent phase (Rp) was considerably faster in the cord ghosts. Fatty acid analysis in the membrane lipids showed that the active bisallylic hydrogen (active H) content was greater in cord ghosts than in adult ghosts. The active H content closely correlated with the Rp, but did not with the Rinh. The kinetic chain length (KCL), i.e., the ratio of the rate of propagation to that of initiation, was calculated from Rp and tocopherol consumption rate and KCL values were higher in cord ghosts than in adult ghosts. The faster Rp and the higher KCL of the cord ghosts were attributable to a greater active H content rather than to the tocopherol content.

Adult↗

Production of hydroxyl radicals by tumor cells varies with cell type as measured by electron spin resonance spectrometry.

The hydroxyl radicals produced by two adherent cell lines, a human cancer cell and a mouse fibroblast, and six suspended human leukemia cell lines at different stages of differentiation were measured by electron spin resonance (ESR) spectrometry. The concentration of hydroxyl radicals detected in these tumor cells increased in proportion to temperature and cell number. The addition of the enzymes superoxide dismutase (SOD) and catalase (which do not permeate the cell membrane), reduced the amount of hydroxyl radicals detected. SOD decreased hydroxyl radicals somewhat but catalase eliminated hydroxyl radicals almost completely. These findings suggest that hydroxyl radicals are produced extracellularly consisted primarily of H2O2 but partially from superoxide radicals. Using the human leukemia cell lines at different stages of differentiation we demonstrated that cell differentiation may correlate with hydroxyl radical production. The earlier the stage of leukemic cell differentiation the more the greater the production of hydroxyl radicals. Moreover, the ability of SOD or catalase to eliminate hydroxyl radical activity correlated inversely with leukemic cell differentiation.

Animals↗

[Monoclonal antibody MIA15-5, which inhibits cell motility and metastatic potential, has a great effect on the prognosis of the post-surgical patients with lung cancer].

The functional monoclonal antibody, selected based on the inhibition of cell motility, was found to be directed to specific carbohydrate structure Fuc alpha 1----2 Gal beta 1----R. This monoclonal antibody, MIA15-5 was established after immunization of mice with adenocarcinoma line of the lung PC7, and selected based on inhibition of U937, HEL, and MAC10 cell migration due to the transwell assay. MIA15-5 reacted with 30-40% of high metastatic variant BL6 of mouse melanoma B16 line and metastatic deposition to lung after injection of BL6 cells was strongly inhibited if MIA15-5 was injected within 3 hours, but was not inhibited by injection of other anti-H antibodies under the same conditions. Injection of MIA15-5 had much less inhibitory effect if administered 1 day after BL-6 injection, and no effect at 3 days. In immunohistochemical staining of 149 formalin fixed specimens of patients with lung cancer, especially adenocarcinoma has the highly frequency of MIA15-5 (65.4%, 49 of 75 cases) and the five year survival rate of the patients with negative staining of MIA15-5 was 58.6% and much better than one with positive staining, 20.9%.

Adenocarcinoma↗

[A case of paragonimiasis Miyazakii with bilateral pleural and pericardial effusion].

A 38-year-old man was admitted to our hospital for further examination of bilateral pleural and pericardial effusion. He had complained of dyspnea on exertion occurring six months after ingestion of raw freshwater crabs, Patomon dehaani. X-ray films and CT scan of the chest taken on admission revealed massive bilateral pleural and pericardial effusion. High serum IgE level and eosinophilia were noted on laboratory examination. Antibody against Paragonimus Miyazakii antigen was detected in patient's serum as well as pleural and pericardial fluids by the Ouchterlony test. The patient was treated effectively by Bithionol.

Adult↗

[Human neuroblastoma cell lines having smooth muscle cell markers].

The neural crest gives rise to a variety of tissues, including peripheral neurons, Schwann cells, melanocytes and ectomesenchymal cells, which include the smooth muscle cells of large arteries. Cell lines derived from neuroblastoma (a neural crest tumor) have at least two distinct morphological cell types, a neuroblastic phenotype (N-type) and an epithelial-like phenotype (S-type) with characteristics of substrate-adhesiveness. We have analyzed 17 human neuroblastoma cell lines using a panel of monoclonal antibodies against cytoskeletal proteins. Three neuroblastoma cell lines (KP-N-SI, KP-N-YN and SMS-KCN) bound an alpha -smooth muscle actin antibody. In addition, one of these cell lines (KP-N-SI) bound anti-desmin monoclonal antibodies as determined by indirect immunofluorescence. A total of eight cloned cell lines were obtained from the above parent cell lines. These were composed of either N- or S-type cells and were confirmed to be the common neuroblastoma origin from each parent cell line by chromosomal analysis. Alpha-smooth muscle actin and desmin were demonstrated in the S-type cloned cells by indirect immunofluorescence, as well as by two-dimensional Western blot analysis. These results were confirmed by Northern blot analysis using a specific probe (pSH alpha SMA-3'UT) to human alpha-smooth muscle actin mRNA. This is the first report of the presence of alpha-smooth muscle actin and desmin in neuroblastoma cell lines. These data show that in addition to giving rise to cells with neural, Schwann cell and melanocyte markers, neuroblastoma can also give rise to the cells expressing smooth muscle cell markers.

Actins↗

High concentration of free trimethyllysine in red blood cells.

A high concentration of a basic unidentified amino compound was found in the blood of rats. It was isolated and identified as N epsilon,N epsilon,N epsilon-trimethyllysine by paper chromatography, thin-layer chromatography, high-performance liquid chromatography and amino acid analyzer. It was localized exclusively in red blood cells in the blood of rats. Free trimethyllysine was also determined in the liver, kidney, spleen, brain, muscle, heart and testis of rat. The concentration of free trimethyllysine in red blood cells was more than 10-times as high as that in the other tissues. This compound in red blood cells was found in different species of animals. The relationship between this free trimethyllysine and carnitine was discussed.

Amino Acids↗

The abnormal occurrence and the differentiation-dependent distribution of N-acetyl and N-glycolyl species of the ganglioside GM2 in human germ cell tumors. A study with specific monoclonal antibodies.

Human primary germ cell tumors were analyzed for the presence of the ganglioside GM2 using three specific monoclonal antibodies which can distinguish the molecular species of the sialic acid moiety: the antibody MK1-16 is specific for N-acetyl GM2, MK2-34 is specific for N-glycolyl GM2, and MK1-17 detects both N-acetyl and N-glycolyl GM2. When the occurrence of the GM2 antigen was tested in 107 cases of human germ cell tumors by the immunohistochemical technique using these antibodies, seminoma was characterized as having the highest frequency of N-acetyl GM2 (89.4%, 42 of 47 cases) among germ cell tumors, followed by embryonal carcinoma (40.0%), and teratocarcinoma (26.6%). Compared with this, yolk sac tumors and choriocarcinoma had a much lower positive incidence of the N-acetyl GM2 antigen. On the other hand, the N-glycolyl GM2 antigen was not found at all in 47 cases of seminoma (0%), and the positive incidence was very low in embryonal carcinoma (6.6%), although considerably higher incidences were obtained with choriocarcinoma (25.0%), yolk sac tumor (22.2%), and teratocarcinoma (13.3%). The presence and molecular species of the GM2 antigens in these human germ cell tumors were also ascertained chemically by the thin-layer chromatography (TLC) immunostaining of the ganglioside fractions prepared from primary germ cell tumors. These results indicate that seminoma specifically contains N-acetyl GM2 and no N-glycolyl GM2, suggesting that N-acetyl GM2 could be a good marker for seminoma. On the other hand, non-seminomatous germ cell tumors were characterized by the presence of N-glycolyl GM2, one of the Hanganutziu-Deicher antigens (H-D antigens). Moreover, the positive occurrence of N-glycolyl GM2 correlated very well with the degree of differentiation of non-seminomatous germ cell tumors, i.e., the differentiated tumors such as yolk sac tumors, choriocarcinoma, and teratocarcinoma had a higher positive incidence of N-glycolyl GM2 type H-D antigen but a lower positive incidence of N-acetyl GM2 when compared with embryonal carcinoma, the most undifferentiated tumors among non-seminomatous germ cell tumors.

Antibodies, Monoclonal↗

The endogenous induction of tumor necrosis factor serum (TNS) for the adjuvant postoperative immunotherapy of cancer--changes in immunological markers of the blood.

The endogenous induction of tumor necrosis factor serum (TNS) for cancer immunotherapy was undertaken in the immediate postoperative period using Lentinan as the primer and OK-432 as the inducer. The changes in several immunological markers of the blood were assayed and compared with a control group to clarify the effects of this treatment. Plasma TNF-alpha levels were elevated two to three hours after eliciting treatment. The neutrophil count was elevated on the 7th postoperative day (POD) and the natural killer (NK) cell activity was transiently suppressed on the 1st POD, but NK cells possessing a high activity (Leu7-CD16+) were preserved until the 7th POD. Helper/inducer (CD4+) and killer cells (CD8+ CD11-) tended to increase, and suppressor (CD8 bright+ CD11+) cells tended to decrease in the induction group. There was no difference in the levels of prostaglandin E2 (PGE2) between the groups, but a marked elevation of interferon-gamma was evident on the 1st POD in the induction group. This treatment may be useful as postoperative adjuvant immunotherapy for cancer due to its ability to induce cytokines and activate host immune mechanisms.

Aged↗

Biooxidation of a synthetic waste by a microbial film grown on the liquid surface in a shallow flow reactor.

A new process of biological waste treatment was developed by use of microbial films grown on the liquid surface in a shallow flow reactor. The performance of this process was tested using a synthetic waste that contained acetic acid as a model organic pollutant. About 90% of acetic acid (10,000 mg/L-1) in the synthetic waste was removed by setting alpha tau: (alpha specific liquid surface area, cm-1, and tau: hydraulic liquid detention time, h) higher than 15 cm-1/h. It was necessary to maintain alpha large enough (more than 10 cm-1 in most cases) to satisfy oxygen demand for the biooxidation. The oxygen balance and TOC determinations showed that the acetic acid removed was completely oxidized with oxygen transferred through the liquid surface. This process would contribute to energy savings, since it requires no forced aeration for oxygen supply.

Acetates↗

Effect of haloperidol on immunoreactive neuropeptide Y in rat cerebral cortex and basal ganglia.

To clarify the dopaminergic regulation of neuropeptide Y (NPY) neurons, the effect of haloperidol on NPY in basal ganglia and the cerebral cortex of the rat brain was investigated by sensitive radioimmunoassay and immunocytochemistry using antiserum against rat NPY. After repeated intraperitoneal injections of haloperidol (5 mg/kg) for 6 days, the content of immunoreactive NPY was significantly decreased in the caudate-putamen, but significantly increased in the lateral prefrontal cortex. After treatment for 21 days, the content of immunoreactive NPY in the caudate-putamen remained significantly low, but the extent of change in the lateral prefrontal cortex diminished. In the medial prefrontal cortex, piriform cortex, parietal cortex and nucleus accumbens, no significant changes were found after treatment for either 6 or 21 days. These findings were compatible with those obtained by immunocytochemistry using the same antiserum: an increase of immunoreactive fibers and terminals in the lateral prefrontal cortex and their decrease in the caudate-putamen. However, in the nucleus accumbens the density of immunoreactive fibers and terminals was decreased in the rostral portion, but not in the caudal portion after haloperidol treatment for 6 and 21 days. These findings suggest that dopaminergic afferents region-specifically regulate dopamine-sensitive NPY neurons in the rat brain.

Animals↗

Increased mitochondrial uptake of rhodamine 123 during interferon-gamma stimulation in Molt 16 cells.

The positively charged rhodamine analog rhodamine 123 accumulates specifically in the mitochondria of living cells. Mitochondria-specific interaction of this molecules is apparently dependent on the high transmembrane potential maintained by functional mitochondria (1, 2, 3). The application of such potential-dependent probes in conjunction with flow cytometry allows the monitoring of mitochondrial membrane potential in living cells (4, 5, 6). In the present work, the uptake of rhodamine 123 by Molt 16 cells stimulated by IFN-gamma or IFN-beta was measured by flow cytometry. Marked elevations in mitochondria-associated probe fluorescence have been observed in cells as early as 30-60 min after exposure of IFN-gamma but not observed by IFN-beta exposure. This results suggest that the IFN-gamma and -beta induced biological functions are different in the control of energy metabolism and energy requirements at the cellular level.

Biological Transport, Active↗

Human IgG3 monoclonal antibody directed to an unbranched repeating type 2 chain (Gal beta 1----4GlcNAc beta 1----3Gal beta 1----4GlcNAc beta 1----3Gal beta 1----R) which is highly expressed in colonic and hepatocellular carcinoma.

Previously established human monoclonal antibodies (MAbs) directed to carbohydrate antigens are essentially all IgM class, and show relatively low affinity and low reactivity at 37 degrees C. We report here the establishment of a human IgG3 MAb displaying high affinity antigen-binding activity at 37 degrees C and efficiently activating cellular cytotoxicity directed to human tumor cell lines expressing the polylactosamine antigen. The IgG3 MAb (MH21-134) reacted with the repeated unbranched polylactosamine structure Gal beta 1----4GlcNAc beta 1----3Gal beta 1----4GlcNAc beta 1----3Gal beta 1----R, i.e., nLc6, nLc8, etc., but did not react with sialyl 2----3 or 2----6 substituted derivatives at the terminal Gal. This specificity differs from that of several anti-i antibodies, or human anti-i-like MAbs which react with sialyl 2----3 substituted structures. Directly biotinylated MH21-134 antibody was used in immunohistochemical staining of 154 formalin-fixed, paraffin-embedded tissue sections to study distribution of the antigen. High incidence of positive staining was found in colon cancer (11/17; 65%) and hepatocellular carcinoma (8/12; 67%), followed by large cell and squamous cell carcinoma of lung cancer (10/13; 59%, and 14/26; 54%, respectively). TLC immunostaining of glycolipid extracts from a variety of tumor tissues showed the presence of nLc6 and/or nLc8 in over 50% of cases. The antigens nLc6 and nLc8 were found to be absent from normal colonic epithelia, kidney, and pancreas. Only a weak band corresponding to nLc8 and one corresponding to nLc6 were found in liver and spleen, although all these normal tissues, including gastrointestinal epithelia, lung, liver, spleen, erythrocytes, and lymphocytes, were essentially negative on immunohistology. However, the antigen was found to be highly expressed in myelocytes and weakly in bronchial glands of lung and pancreatic duct epithelia. Nevertheless, expression of unsubstituted, unbranched polylactosamine antigen could be an important basis for induction of humoral immune response against certain types of human cancer, despite its limited expression in normal cells.

Amino Sugars↗

Production of identical twins by separating two-cell rat embryos.

Rat identical twins were produced from two-cell embryos. In the presence of cytochalasin B, rat two-cell embryos could be separated efficiently into two blastomeres by micromanipulation. Isolated blastomeres, embedded in agar cylinders and cultivated in ligated rat oviducts for 3 days, developed to the morula or blastocyst stage. After removing the agar, pairs of developed one-half embryos were transferred into Day 1 oviducts or Day 4 uteri of pseudopregnant rats. The percentage of embryos, separated either in the presence or absence of cytochalasin B, that developed into live fetuses was higher in cases of uterine transfer than in cases of oviduct transfer (38% vs. 18%, 31% vs. 15%, respectively). Throughout the present experiment, nine pairs of identical twins were successfully produced. This is the first report of the production of identical rat twins by separating two-cell embryos.

Animals↗

Motility and fertilizing ability of rat epididymal spermatozoa washed by a continuous gradient of Percoll.

Removal of epididymal fluids from epididymal sperm suspension is an important step for the study of sperm motility, capacitation, and the acrosome reaction. The technique of washing should minimize damage to viable spermatozoa but at the same time efficiently remove debris, non-sperm cells, and biological fluids. We examined sperm motility and fertilizability in vitro of rat epididymal spermatozoa after washing with Percoll continuous gradient. Nine milliliters (ml) of 50% N-2-hydroxyethylpiperazine-N1-2-ethanesulfonic acid (HEPES) buffered Percoll solution was centrifuged at 20,000 g for 45 minutes to form a continuous gradient. One hundred to 300 microliters of sperm suspension was loaded onto the surface of the gradient and centrifuged at 150 g and 1,500 g for 10 minutes. Two main layers of spermatozoa were formed, one of high (lower layer) and one of low (upper layer) motility. At centrifugation 1,500 g, the sperm density and motility in the lower layer were greater than at 150 g. Spermatozoa from both layers at 150 g and at 1,500 g were diluted with modified Krebs-Ringer's bicarbonate solution (mKRB) and preincubated for 5 hours. Superovulated eggs collected from 21-25-day-old Wistar strain immature rats were introduced into the preincubated sperm suspension for insemination and fixed 5-5.5 hours later for observation of fertilization. Spermatozoa from both layers, 150 g and 1,500 g, showed the same fertilizability in vitro as control spermatozoa. From these results we conclude that Percoll gradients can be used for washing rat epididymal sperm for the study of sperm physiology including fertilization.

Animals↗

Single potassium channels opened by opioids in rat locus ceruleus neurons.

Currents through single-ion channels were recorded in the cell-attached configuration from locus ceruleus neurons enzymatically dissociated from newborn rats. When the selective mu opioid receptor agonist Tyr-D-Ala-Gly-MePhe-Gly-ol was in the patch-clamp electrode, unitary inward currents were observed with conductance of approximately 45 pS (measured at zero pipette potential, with 150 mM potassium in the recording electrode). Long silences, lasting many seconds to minutes, separated periods of activity of similar durations. Within such activity periods the distribution of closed times of the channels was best fitted by the sum of two exponential functions (time constants approximately 1 and 30 ms), and the durations of channel openings were fit by a single exponential function; mean open time increased from 2 to 120 ms as agonist concentration increased. Channel activity was not seen when high concentrations of opioids were applied to the neuron outside the patch-clamp recording electrode, indicating intimate coupling between receptor and potassium channel. Unitary currents with similar properties were also seen when pipettes contained alpha 2 adrenoceptor agonists or somatostatin. Taken with previous findings, the results indicate that mu opioid receptors, alpha 2 adrenoceptors, and somatostatin receptors can couple directly to membrane potassium channels through the local intermediary action of a GTP binding protein.

Action Potentials↗