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Biomedical subjects

C Tanaka

Publications and source records attributed to C Tanaka.

At least 325 records · Page 18Linked to original sources

Radiofrequency hyperthermia with successive monitoring of its effects on tumors using NMR spectroscopy.

Radiofrequency (rf) hyperthermia was generated on rat glioma inoculated s.c. in CD Fisher rats by applying the rf pulse using the surface coil in the NMR spectrometer, and the effect was monitored successively in the same spectrometer by measuring 31P NMR spectra and 1H NMR images. In the 31P NMR spectrum at the preirradiation stage, nucleoside triphosphate peaks and a phosphomonoester peak were high and a Pi peak was low. After a rf pulse at a power of 5 W was applied continuously for 60 min, the nucleoside triphosphate peaks decreased and the Pi peak increased immediately, resulting finally in a dominant Pi peak pattern within 30 min in all 10 cases examined. These spectral changes occurred much earlier than the histological changes and lasted for at least 7 days. By the 1H NMR imaging, the necrotic region was detected as a high-intensity lesion in spin echo and inversion recovery images 2 days after the irradiation. There were no changes either in the spectrum or in 1H NMR images in any of 8 cases after irradiation with a rf pulse of less than 3 W. Thus, we could generate rf hyperthermia with the NMR spectrometer and the effects were monitored sensitively with the same spectrometer. It can be concluded that the NMR device can be used not only for diagnosis but also as a therapeutic tool.

Animals↗

Presynaptic muscarinic and alpha-adrenoceptor-mediated regulation of GABA release from myenteric neurones of the guinea-pig small intestine.

The effects of cholinomimetic and sympathomimetic drugs on the release of [3H]-gamma-aminobutyric acid ([3H]-GABA) evoked by high K+ from the isolated small intestine of the guinea-pig were investigated, in the presence of tetrodotoxin. Acetylcholine and oxotremorine, at concentrations ranging from 10(-9) to 10(-6) M inhibited the evoked release of [3H]-GABA in a concentration-dependent manner, while nicotine was without effect. Scopolamine and pirenzepine inhibited the effect of oxotremorine, while hexamethonium had no effect. The IC50 values for scopolamine and pirenzepine of the oxotremorine (3 X 10(-8) M)-induced inhibition were 1.02 X 10(-9) M and 9.78 X 10(-10) M, respectively. Noradrenaline, but not isoprenaline inhibited the evoked release of [3H]-GABA. Clonidine (10(-10)-10(-6) M) reduced the evoked release of [3H]-GABA in a concentration-dependent manner, but phenylephrine had no effect. The inhibitory effect of clonidine was antagonized by yohimbine but not by prazosin. These findings provide evidence for the localization of M1-muscarinic and alpha 2-adrenoceptors on GABAergic nerve terminals and their involvement in the presynaptic control of the release of GABA from the guinea-pig small intestine.

Animals↗

Evaluation of the effects of photoradiation therapy on brain tumors with in vivo P-31 MR spectroscopy.

In vivo phosphorus-31 magnetic resonance (MR) spectra were obtained by a surface coil method from rat glioma tissue inoculated subcutaneously in CD Fisher rats, and the effects of photoradiation therapy on tumors were evaluated by sequentially observing spectral changes. In the control group, the nucleoside triphosphate (NTP) and phosphomonoester peaks were large, the phosphocreatine peak was small, and the inorganic phosphate (Pi) peak was intermediate. In all eight cases in the group in which a dose of 10 mg/kg of hematoporphyrin derivatives (HpD) was given before photoirradiation, NTP peaks decreased, and the Pi peak increased remarkably within 1 hour after the 60-minute white-light irradiation. Spectral changes were observed before histologic changes were apparent. Histologic examinations 3 days after irradiation showed extensive necrosis in the tumor tissue. With preinjection of 5 mg/kg HpD, three of the eight cases showed spectrum changes after the irradiation. No spectrum changes were observed in the group with preinjection of 2.5 mg/kg. In vivo P-31 MR spectra measurements are useful not only to investigate the energy metabolism of tumor tissue in vivo but also to evaluate the effects of photoradiation therapy on tumors.

Animals↗

Activation of protein kinase C potentiates norepinephrine release from sinus node.

Localization of binding sites of [20-3H]phorbol-12,13-dibutyrate [( 3H]PDBu) and the involvement of Ca2+-phospholipid-dependent protein kinase (protein kinase C) in the release of norepinephrine (NE) from sympathetic nerve terminals in the guinea pig sinus node were investigated. There was a single class of specific [3H]PDBu binding sites in the heart. [3H]NE release from the sinus node preloaded with [3H]NE was evoked by electrical stimulation in superfusing medium containing Ca2+ or by the concomitant presence of Ca2+ ionophore and Ca2+, in Ca2+-free medium. 12-O-tetradecanoylphorbol-13-acetate (TPA) potentiated the evoked [3H]NE release. The effect of TPA was antagonized by both polymyxin B and H-7, inhibitors of protein kinase C. TPA increased the apparent affinities of electrical stimulation-evoked release for extracellular Ca2+. The possibility that protein kinase C plays a role in transmembrane signal transduction involved in the release of NE from peripheral adrenergic nerve terminals in the guinea pig sinus node warrants continued study.

Animals↗

Vasoactive intestinal polypeptide provokes acetylcholine release from the myenteric plexus.

Effects of vasoactive intestinal polypeptide (VIP) on the release of acetylcholine (ACh) from longitudinal muscle strips with myenteric plexus (LM) preparations were examined in the guinea pig small intestine. VIP (10(-10) to 10(-6) M) induced a concentration-dependent contraction of LM preparation. The VIP-induced contractions seem to be related to three components, the scopolamine-sensitive, the scopolamine-insensitive, the tetrodotoxin-sensitive, and the tetrodotoxin-insensitive contractions. VIP (10(-10) to 10(-6) M) induced a concentration-dependent increase in the release of [3H]ACh from LM preparations preloaded with [3H]choline. The VIP-evoked [3H]ACh release was inhibited by removal of Ca2+ from the perfusion medium and by treatment with tetrodotoxin but not by scopolamine and hexamethonium. The spontaneous and VIP-evoked [3H]ACh release was not affected by phentolamine, propranolol, methysergide, diphenhydramine, cimetidine, bicuculline, or [D-Pro2, D-Trp7,9]substance P. The result demonstrates that VIP induces contractions of longitudinal smooth muscle directly and indirectly by the stimulation of both cholinergic neurons and noncholinergic excitatory neurons.

Acetylcholine↗

[Epinephrine uptake and release and presynaptic alpha 2-adrenoceptic regulation in the guinea pig hypothalamus].

The uptake and the release of [3H] epinephrine ([3H] E) and the release of endogenous E in slices of guinea pig hypothalamus were investigated. [3H]E rapidly accumulated in the slices incubated in Krebs-Ringer solution containing [3H]E. Kinetic analysis indicated two components of E accumulation, one representing a high (Km1, 7.7 X 10(-8) M and Vmax1, 0.13 pmoles/mg/10 min) and the other a low (Km2, 1.8 X 10(-6) M and Vmax2, 1.4 pmoles/mg/10 min) affinity uptake system. Endogenous E released in response to electrical stimulation was estimated using gas chromatography and mass spectrometry. The electrical stimulation produced a release of both [3H]norepinephrine ([3H]NE) and [3H]E from hypothalamic slices preloaded with [3H]NE. With electrical stimulation of the slices, there was an efflux of [3H]E from tissues preloaded with [3H]E, in a current- and frequency-dependent manner. Electrically stimulated release of [3H]E from the slices was inhibited by tetrodotoxin (10(-6) M) and by a calcium-free medium containing EGTA (10(-4) M), in cases of up to 1 mA of intensity of electrical stimulation. The release of [3H]E induced by electrical stimulation was enhanced by yohimbine, and this effect was suppressed by clonidine. These results provide strong evidence for the neurotransmitter role of this catecholamine in the hypothalamus and suggest the possible existence of a presynaptic regulatory mechanism of E release, through the presynaptic alpha 2 receptors on the E nerve terminals.

Animals↗

Dopamine release and presynaptic dopaminergic regulation in guinea pig spinal cord.

Using pharmacological approaches, we obtained evidence for the release of dopamine (DA) and its dopaminergic regulation in the guinea pig spinal cord. Electrical stimulation of the cord pieces increased the endogenous DA release and the efflux of [3H]dopamine [( 3H]DA) from tissues preloaded with [3H]DA. The evoked release of [3H]DA was current- and frequency-dependent and was prevented by tetrodotoxin (10(-6) M) or Ca2+-free medium containing EGTA (10(-4) M), while benztropine allowed a recovery of a more extensive amount of [3H]DA in the superfusing medium by inhibiting the reuptake process. The stimulated [3H]DA release was reduced by LY-171555, but not by SKF-38393 and 8-Br-cAMP. (-)Sulpiride enhanced the stimulated endogenous DA and [3H]DA release. (-)Sulpiride reversed the inhibition of evoked [3H]DA release induced by LY-171555, while SKF-38393 and 8-Br-cAMP did not affect LY-171555-induced inhibition of evoked [3H]DA release. These findings provide additional evidence for the neurotransmitter role of DA in the spinal cord of the guinea pig, and they strongly suggest the presence of presynaptic regulation of DA release via the dopamine receptor which is the D2 type.

Animals↗

Recognition of serum alkaline phosphatase by murine monoclonal antibodies against human osteosarcoma cells.

Using OST6 and OST7 monoclonal antibodies against human osteosarcoma cells, a solid-phase radioimmunosandwich assay was developed to quantitate a human osterosarcoma-associated antigen in a total of 242 sera from healthy adults and patients with various diseases. The levels of the antigen in sera were high in patients with osteosarcoma and in children without tumorous diseases compared with healthy adults; however, the highest level of the antigen was found in patients with obstructive jaundice. The quantity of the antigen correlated with serum alkaline phosphatase (EC 3.1.3.1.) activity, and showed a strong correlation (correlation coefficient, 0.94) in 50 sera. Immunolocalization of enzyme activity assay using monoclonal antibodies was performed to ascertain whether the antigen had alkaline phosphatase activity. This assay proved that OST6, OST7, and OST15 monoclonal antibodies recognized serum alkaline phosphatase; furthermore, these monoclonal antibodies seemed to react with not only the bone isoenzyme but also the liver isoenzyme.

Alkaline Phosphatase↗

gamma-Aminobutyric acid in peripheral tissues.

Significant amounts of gamma-aminobutyric acid (GABA), an endogenous amino acid, are present in mammalian peripheral tissues. This finding led to the suggestion that GABA may act as a neurotransmitter in the peripheral nervous system as it does in the central nervous system. This review deals with recent identification of GABA in the autonomic nervous system and the possible functional role of GABA in neuronal and non-neuronal tissues. The identification of GABA in the autonomic nervous system has paved the way for new approaches in pharmacological investigations.

Animals↗

Target size analysis of serotonin 5-HT1 and 5-HT2 receptors in bovine brain membranes.

Freeze-dried crude synaptic membranes prepared from bovine cerebral cortex and striatum were exposed to high energy gamma ray from the source of 60Co. The size of serotonin 5-HT1 receptors labeled by [3H]serotonin and that of 5-HT2 receptors labeled by [3H]spiperone or [3H]ketanserin was determined by target size analyses. The values were 57,000 daltons, 145,000 daltons and 152,000 daltons for the cerebral cortex and 56,000 daltons, 141,000 daltons and 150,000 daltons for the striatum, respectively. The estimated sizes were deduced by reference to enzyme standards with known molecular masses and which were irradiated in parallel. Our results demonstrate that the molecular entities in situ for 5-HT1 receptors are distinct from those for 5-HT2 receptors, thus supporting data on the existence of two distinct populations of serotonin receptors, hitherto evidenced physiopharmacologically.

Animals↗

Histamine H1-receptors in the guinea-pig urinary bladder.

Histamine H1-receptors were identified in the guinea-pig urinary bladder. Histamine (10(-8)-10(-3) M) produced a dose-dependent contraction which was not altered by either scopolamine or tetrodotoxin. Specific [3H]mepyramine binding to the urinary bladder was saturable and there was a single population of high affinity binding sites with an equilibrium dissociation constant (KD) of 0.77 nM and maximum binding capacity (Bmax) of 68.5 fmol/mg protein. Histamine-induced contraction and [3H]mepyramine binding were inhibited by triploridine, promethazine, d-chlorpheniramine and mepyramine but not by cimetidine. There was a good correlation between mechanical activity and [3H]mepyramine binding as shown by the inhibition affinity constant (Ki) of H1-antagonists. These results provide evidence that the histamine H1-receptor in the guinea-pig urinary bladder is involved in the histamine-induced contraction.

Animals↗

Effect of hypoxia on uptake and acetylation of [3H]choline in brain slices from adult and newborn guinea pigs.

The synthesis of [3H]acetylcholine in brain slices from adult and newborn guinea pigs were suppressed by anaerobiosis. The uptake and acetylation of [3H]choline in the newborn brain were very low, compared to findings in slices of the adult brain. The most sensitive tissue to hypoxia was the striatum followed by the hippocampus. There were significant differences in the case of the frontal cortex.

Acetylation↗

Regional distribution of [3H]nitrendipine binding in human brain.

Evidence for the binding of a calcium antagonist, [3H]nitrendipine, to human brain membranes was obtained. This binding was saturable, specific and of high affinity with a dissociation constant (Kd) of 0.62 nM in the prefrontal cortex and 0.82 nM in the caudate. The maximal numbers of binding sites (Bmax) in these areas were 57.5 and 32.3 fmol/mg protein, respectively. [3H]Nitrendipine binding to membranes from 27 regions in the post-mortem human brains was measured. The highest levels of 0.5 nM [3H]nitrendipine binding were seen in the cerebral cortex, amygdala and thalamus, and levels in the midbrain, cerebellum and pallidum were considerably lower.

Adult↗

Molecular size of histamine H-1 receptor determined by target size analysis.

Target size analysis (radiation inactivation) was used to study the molecular size of the histamine H-1 receptor of bovine and human cerebral cortex in the intact membrane-bound state. The H-1 receptor in bovine and human cerebral cortex was found to exist in the membrane as a homogeneous population of the same size (160,000 daltons) in each case. Thus no evidence for the existence of multiple forms of the receptor has been found.

Animals↗

Autoreceptors regulate gamma-[3H]aminobutyric acid release from the guinea pig small intestine.

The possible presence of a presynaptic gamma-aminobutyric acid (GABA) receptor capable of regulating the release of GABA was investigated using the guinea pig small intestine. Muscimol at 10(-8) M and 10(-7) M, but not baclofen at 10(-6) M, inhibited the K+ (40 mM)-evoked Ca2+-dependent release of [3H]GABA from the small intestine preloaded with [3H]GABA, in the presence of 10(-6) M tetrodotoxin. The effect of muscimol on the K+-evoked GABA release was inhibited by bicuculline and furosemide. These results show that the guinea pig small intestine possesses presynaptic GABA receptors which may be involved in the regulation of the evoked GABA release. The presynaptic GABA receptor is bicuculline-sensitive and is probably coupled to the Cl- ion channel.

Animals↗

Epinephrine release and presynaptic alpha 2-adrenoceptic regulation in the guinea pig hypothalamus.

The release of epinephrine (E) and its regulation system were investigated in the slices of guinea pig hypothalamus. Electrical stimulation produced an efflux of [3H]E from tissues preloaded with [3H]E, in a current- and frequency-dependent manner. Stimulated [3H]E release was inhibited by tetrodotoxin and by a calcium-free medium containing ethylene glycol bis(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA). Stimulated [3H]E release was enhanced by yohimbine and this effect was suppressed by clonidine. These results provide strong evidence for the neurotransmitter role of E in the hypothalamus, and suggest the possible existence of regulatory mechanism of E release via presynaptic alpha 2-receptors.

Animals↗

[125I] radioiodinated metaraminol: a new platelet-specific labeling agent.

In our search for a platelet-specific labeling agent, metaraminol (MA), a low-toxic pharmaceutical for the treatment of hypotension and cardiogenic shock, attracted our attention. Its active incorporation and accumulation by platelets have been recognized. At first, the preparation of 125I radioiodinated metaraminol (125I-MA) was carried out using the chloramine-T method. Then, upon the harvest of platelets as platelet-rich plasma (PRP), their labeling with this new radiopharmaceutical was easily performed by incubation for 10 min at 37 degrees C. The cell-labeling efficiency was dependent on cell density, reaching 63.0% +/- 3.1% at 2.4 X 10(9) cells/ml. The specific incorporation of 125I-MA by an active transport system similar to that of 5-hydroxytryptamine (5-HT) as well as by passive diffusion was demonstrated. In in vitro studies, the unaltered state of 125I-MA-labeled platelets with their cellular functions fully retained was estimated. In vivo studies carried out in rabbits with induced thrombi in the femoral artery showed a rather rapid disappearance of the radioactivity from circulating blood, reaching a high thrombus-to-blood activity ratio of 19.8 +/- 4.3 within 30 min of the administration of 125I-MA-labeled autologous platelets. Thus, with the potential availability of 123I, 123I-MA-labeled platelets appear to be a promising agent for thrombus imaging using single-emission computed tomography (CT) studies.

Animals↗