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T Akasu

Publications and source records attributed to T Akasu.

At least 73 records · Page 4Linked to original sources

Detection of K-ras point mutations in mesenteric venous blood from colorectal cancer patients by enriched polymerase chain reaction and single-strand conformation polymorphism analysis.

In order to confirm the presence of cancer cells in mesenteric venous blood and to examine their relationship with the occurrence of liver metastases, we attempted to detect K-ras codon 12 point mutations in perioperative mesenteric blood using enriched polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) analysis in 25 patients with primary colorectal tumors carrying K-ras point mutations. Among these patients, three with synchronous liver metastases were included. The same K-ras point mutation (substitution of GAT for GGT) was detected in both the blood and the primary tumor in a Dukes' C patient. We confirmed this result by colony hybridization and estimated the tumor-to-normal cell ratio to be 1:400. This patient has no liver metastases two years after surgery and her carcinoembryonic antigen (CEA) level remains normal. We demonstrated that considerable numbers of cancer cells can be found in mesenteric venous blood during colorectal cancer surgery. However, their potential role in the formation of liver metastases remains unclear.

Colorectal Neoplasms↗

Muscarine activates a nonselective cation current through a M3 muscarinic receptor subtype in rat dorsolateral septal nucleus neurons.

1. In the present study, we examined the cellular mechanism and receptor type responsible for a muscarine-induced inward current (Imi) in neurons of rat dorsolateral septal nucleus (DLSN) using single-microelectrode voltage-clamp and "slice" patch-clamp techniques. 2. Imi was associated with an increase of membrane conductance in 75% of DLSN neurons. There was no voltage-dependence of Imi between -60 and -140 mV; it exhibited a reversal potential of -17.0 +/- 5.3 mV (n = 14) determined by extrapolation of Imi and voltage relationship recorded using whole cell patch recording. Lowering extracellular sodium (26 mM) or potassium (1.4 mM) ions depressed Imi. 3. Imi was concentration dependent; 3 and 100 microM muscarine produced the minimum [22 +/- 4.6 pA, (mean +/- SE) n = 8] and maximum (167 +/- 28 pA, n = 7) responses, respectively. An EC50 was determined to be 15 microM (n = 8). Oxotremorine-methiodide (1-100 microM) also produced an inward current with similar potency compared with muscarine. On the other hand, McN-A-343 and pilocarpine (3-100 microM) did not produce any inward current in DLSN neurons. 4. Atropine (1 microM) completely reduced Im produced by 30 microM muscarine, whereas pirenzepine (PZP) shifted the concentration-response curve for muscarine in a parallel manner to the right. The EC50 for muscarine was shifted to 32, 52, and 204 microM by 0.2, 0.5, and 2 microM PZP, respectively. The apparent Kd value for PZP estimated by Schild plot analysis was 190 nM (n = 5). 5. Methoctramine (1 microM) also competitively depressed Imi; the calculated EC50 values were 26, 41, and 107 microM in concentrations of 0.2, 2, and 10 microM methoctramine, respectively. The apparent Kd for methoctramine was 420 nM. In contrast, AF-DX 116 (1 microM) did not significantly inhibit Imi. 6. Intracellular dialysis with guanosine 5'-O-(3-thiotriphosphate), a nonhydrolyzable analogue of GTP, suppressed irreversibly Imi. Pretreatment of DLSN neurons with pertussis toxin (PTX) did not prevent Imi (n = 8). 7. We suggest that muscarine causes this inward current by activating a M3 subtype of muscarinic receptor, which is coupled to a PTX-insensitive GTP-protein in rat DLSN neurons.

Animals↗

Presynaptic GABAA receptors in vertebrate synapses.

The presynaptic gamma-aminobutyric acidA (GABAA) receptor has long been considered as the site for 'presynaptic inhibition' of synaptic transmission in the central nervous system (CNS). However, recent reports have indicated that the activation of GABAA receptors depolarizes primary afferent neurons and actually facilitates the release of GABAA, noradrenaline, adenosine, and luteinizing hormone (LH) in central and peripheral tissues. Isoguvacine, a GABAA receptor agonist, enhances substance P (SP) release in the spinal cord. GABAB receptor agonists, bu not GABAA receptor agonists, produce behavioral antinociception in the spinal cord. Baclofen does not directly depolarize the postsynaptic membrane, but presynaptically inhibits the activity of dorsal horn neurons. The excitatory postsynaptic potential (EPSP) evoked by stimulation of dorsal root C-fibers is inhibited by baclofen. Baclofen and GABA inhibit SP release from the primary afferent terminals by activating GABAB receptors. The activation of GABAB receptors inhibits calcium currents in neurons of dorsal roots ganglia (DRG). It is likely that the GABAA receptors act as a site for 'presynaptic facilitation' of transmitter release in the CNS.

Animals↗

Muscarine increases a voltage-independent potassium conductance through an M4 receptor in rat dorsolateral septal nucleus neurons.

The direct effect of muscarine on neurons of the rat dorsolateral septal nucleus (DLSN) was examined by using conventional microelectrode and voltage-clamp techniques. Muscarine (1-50 microM) caused a hyperpolarization accompanied by an increase of a voltage-independent potassium conductance. Pirenzepine competitively antagonized the muscarine-induced hyperpolarization with an apparent dissociation constant (Kd) value of 54 nM. Furthermore, intracellular loading with GTP gamma S, a non-hydrolyzable GTP analog, blocked irreversibly the muscarine-induced hyperpolarization. In addition, pretreatment of neurons with pertussis toxin (PTX) prevented the hyperpolarization produced by muscarine. These results suggest that muscarine hyperpolarizes DLSN neurons via a voltage-independent potassium conductance by acting at M4 subtype receptors which are coupled to a PTX-sensitive G-protein in DLSN neurons.

Animals↗

Patterns of recurrence after nerve-sparing surgery for rectal adenocarcinoma with special reference to loco-regional recurrence.

PURPOSE: Since the early 1980s to relieve functional disturbances after rectal excision, we have been performing nerve-sparing surgery for rectal cancer. The aim of this study was to analyze patterns of recurrences, especially concerning causes of local ones. Furthermore, we would like to address the criteria we used in patient selection to effect successful nerve-sparing surgery. METHODS: From 1982 to 1991, 306 patients underwent nerve-sparing operations, which may be categorized into three types: 1) total autonomic nerve preservation (125 cases), 2) complete pelvic nerve preservation (105 cases), and 3) partial pelvic nerve preservation with removal of parasympathetic nerve (79 cases). Single and multivariant regression analyses were conducted to investigate patterns of recurrence, especially causes of local ones. RESULTS: Sixty-five patients (21 percent) developed recurrent tumors, 19 of which (6.2 percent) were local. Using Dukes terms, there were five patients with Dukes A 13 with Dukes B, and 47 (35 percent) with Dukes C stages. Rate of local recurrences was 13 percent in patients with Dukes C tumor. According to single-variant analysis of Dukes C patients, the following factors are thought to influence local recurrences: number of lymph nodes metastases, level of primary growth, and direction of lymphatic spread. Multivariate regression analysis suggested that lymph node metastasis was the most important and influencing factor on local regrowth (P < 0.002). CONCLUSIONS: Compared with local recurrences is so-called extended surgery appeared to be lower. Our current policy is aggressive application of nerve-sparing surgery, even to patients with node-positive rectal cancer, taking into consideration the exact extent of cancer spread. From the viewpoint of neuroanatomy related to mesorectum, we discussed patient determination for our nerve-sparing surgery.

Adenocarcinoma↗

Inhibition by wortmannin of M-current in bullfrog sympathetic neurones.

1. The actions of wortmannin, an inhibitor of myosin light chain kinase (MLCK), on M-type potassium current of dissociated bullfrog sympathetic neurones have been examined. 2. The amplitude of M-current was measured by whole cell recordings from cells pretreated with wortmannin (0.01-10 microM) or the wortmannin vehicle, dimethylsulphoxide (0.0001-0.1 vol%), for 30 min. Internal (recording pipette) solutions having three different pCa values (6, 7 and 8) were used for the measurements. 3. Irrespective of the pCa, M-current was not detectable when the cells were pretreated with 10 microM wortmannin. Wortmannin, 3 microM, produced 85-95% inhibition of the M-current. Pretreatment with 10-30 nM wortmannin was without effect on M-current. 4. The M-current inhibition by wortmannin at concentrations of 0.1-1 microM depended on the pCa of the internal solution. Inhibition occurred only when the calcium-rich (pCa = 6) internal solution was used. 5. Pre-treatment of the cells with wortmannin (10 microM) did not affect rapidly-inactivating A-type or delayed rectifier-type potassium currents not did it alter inwardly rectifying sodium-potassium current (IH). 6. These observations show that M-current inhibition by wortmannin has two pharmacological profiles. One is calcium-dependent and occurs at lower concentrations (0.1-1 microM), and is attributed to inhibition of MLCK by wortmannin. At higher concentrations (3-10 microM), wortmannin has an additional, calcium-independent action, inhibiting the M-current by an unknown mechanism.

Androstadienes↗

Neurokinin A depolarizes neurons of bullfrog dorsal root ganglia by suppressing K+ conductances.

The effect of neurokinin A (NKA) on neurons of bullfrog dorsal root ganglia (DRG) in primary culture was examined by using whole-cell patch-clamp methods. Application of NKA (1 microM) depolarized the DRG neurons, resulting in spontaneous firing of action potentials. Under voltage-clamp condition, NKA (3 nM-1 microM) caused an inward current (INKA) associated with decreased membrane conductance. The INKA reversed its polarity at the equilibrium potential for K+. The INKA was blocked by extracellular Ba2+ (1 mM) but not by nominally 0 mM Ca2+, tetraethylammonium (40 mM), 4-aminopyridine (2 mM) or apamin (50 nM). Intracellular Cs+ blocked the INKA. NKA depressed a voltage-dependent non-inactivating K+ current, the M-current (IM), at potentials more positive than -55mV. NKA reduced the maximum M-conductance (GM) without changing the kinetics of M-channels. NKA also depressed a voltage- and time-independent background K+ current, IK(B). It is concluded that the INKA is produced by suppression of both IM and IK(B) in bullfrog primary afferent neurons.

Action Potentials↗

cAMP-dependent inward rectifier current in neurons of the rat suprachiasmatic nucleus.

Electrophysiological properties of the inward rectification of neurons in the rat suprachiasmatic nucleus (SCN) were examined by using the single-electrode voltage-clamp method, in vitro. Inward rectifier current (IH) was produced by hyperpolarizing step command potentials to membrane potentials negative to approximately -60 mV in nominally zero-Ca2+ Krebs solution containing tetrodotoxin (1 microM), tetraethylammonium (40 mM), Cd2+ (500 microM) and 4-aminopyridine (1 mM). IH developed during the hyperpolarizing step command potential with a duration of up to 5 s showing no inactivation with time. IH was selectively blocked by extracellular Cs+ (1 mM). The activation of the H-channel conductance (GH) ranged between -55 and -120 mV. The GH was 80-150 pS (n = 4) at the half-activation voltage of -84 +/- 7 mV (n = 4). The reversal potential of IH obtained by instantaneous current voltage (I/V) relations was -41 +/- 6 mV (n = 4); it shifted to -51 +/- 8 mV (n = 3) in low-Na+ (20 mM) solution and to -24 +/- 4 mV (n = 4) in high-K+ (20 mM) solution. Forskolin (1-10 microM) produced an inward current and increased the amplitude of IH. Forskolin did not change the half-activation voltage of GH. 8-Bromo-adenosine 3',5'-cyclic monophosphate (8-Br-cAMP, 0.1-1 mM) and dibutyryl-cAMP (0.1-1 mM) enhanced IH. 3-Isobutyl-1-methylxanthine (IBMX, 1 mM) also enhanced IH. The results suggest that the inward rectifier cation current is regulated by the basal activity of adenylate cyclase in neurons of the rat SCN.

1-Methyl-3-isobutylxanthine↗

Effects of K-7259 on neuronal activity and synaptic transmission in the rat dorsolateral septal nucleus.

K-7259 is a dilazep dihydrochloride derivative that minimizes the damaged area from middle cerebral artery hemiocclusion in the rat (Yamauchi et al. 1992a, b). The effects of K-7259 on the electrophysiological properties of neurons in the rat dorsolateral septal nuclei (DLSN) were examined. K-7259 (100 microM-3 mM) depolarized the membrane with a decrease in input resistance in 36% of the DLSN cells. K-7259 (100 microM) depressed the inhibitory postsynaptic potential (IPSP) and the late hyperpolarizing potential (LHP). The magnitudes of the depressions of the IPSP and LHP with 100 microM K-7259 were 50 +/- 25% (n = 5) and 52 +/- 15% (n = 4), respectively. The amplitudes of the excitatory postsynaptic potentials (EPSPs) were augmented during the inhibition of the IPSP and LHP. However, a voltage-clamp analysis showed that K-7259 did not affect the isolated excitatory postsynaptic current (EPSC). The outward current produced by pressure application of gamma-aminobutyric acid (GABA) to the recording cell was not inhibited by K-7259. These results indicate that K-7259 presynaptically inhibits the IPSP and LHP through a GABAergic pathway.

Animals↗

Patch-clamp analysis of hypoglycemia-induced inhibition of synaptic transmission in the rat dorsolateral septal nucleus.

The role of glucose in neuronal transmission was examined in the rat dorsolateral septal nucleus (DLSN) by using 'slice-patch' techniques. Removal of glucose from the oxygenated Krebs solution produced an outward current associated with an increased K+ conductance. The glucose-depletion depressed the excitatory postsynaptic current (EPSC), the inhibitory postsynaptic current (IPSC) and the late hyperpolarizing current (LHC) produced by stimulation of the fimbria/fornix pathway. It is hypothesized that glucose regulates neurotransmission in the rat DLSN by modulating the release of transmitters from presynaptic nerve terminals.

Animals↗

Intracellular ATP changes the voltage-dependence of delayed rectifier potassium current in bullfrog primary afferent neurons.

Dissociated bullfrog dorsal root ganglion cells were voltage-clamped in the whole-cell configuration to study the steady-state activation and inactivation curves for a delayed rectifier potassium current. The 50%-activation of the current occurred at +15 mV when measured with ATP (5 mM) in the pipette solution as opposed to -11 mV with 5'-adenylylimidodiphosphate (AMP-PNP, 5 mM) and -15 mV with adenosine 5'-O-(3-thiotriphosphate) (5 mM). The 50%-inactivation of the current occurred at -6 mV with ATP but at -31 mM with AMP-PNP. The results suggest that intracellular ATP modulates voltage-dependence of the delayed rectifier in amphibian afferent neurons.

Adenosine Triphosphate↗

Chemosensitivity of C-cells in bullfrog dorsal root ganglia to substance P and adenosine 5'-triphosphate.

Dissociated bullfrog dorsal root ganglion cells were voltage clamped in the whole-cell configuration. In small C-cells having 20 microns as averaged diameter, substance-P (0.1-1 microM) inhibited an M-type potassium current while ATP (1-10 microM) activated a sodium-potassium current. In large A-cells (approximately 65 microns in diameter) in which ATP has been shown to inhibit M-current, substance P (0.1-1 microM) also inhibited this potassium current without activating the sodium-potassium current. Results provided evidence for the distinction between A- and C-cells in terms of their chemosensitivity.

Adenosine Triphosphate↗

Inward rectifier and low-threshold calcium currents contribute to the spontaneous firing mechanism in neurons of the rat suprachiasmatic nucleus.

Intracellular and voltage-clamp studies were carried out to clarify the mechanism for spontaneous firing activity in neurons of the suprachiasmatic nucleus (SCN) of rat hypothalamic brain slices in vitro. SCN neurons displayed spontaneously firing action potentials that were preceded by a depolarizing pre-potential and followed by a short spike after-hyperpolarization (AHP). Injection of inward current with a duration longer than 50 ms resulted in a depolarizing voltage "sag" on hyperpolarizing electrotonic potentials. The inward rectification was depressed by bath application of caesium (1 mM) but not by barium (500 microM). SCN neurons also showed a rebound depolarization associated with spike discharge (anodal break) in response to relaxation of hyper polarizing current injection. The rebound depolarization was reduced by nominally zero calcium. Cadmium (500 microM), cobalt (1 mM) or caesium (1 mM) but not nicardipine also depressed the rebound depolarization. Under voltage-clamp conditions, hyperpolarizing steps to membrane potentials negative to approximately -60 mV caused an inward rectifier current, probably H current (IH), which showed no inactivation with time. Bath application of caesium (1-2 mM) suppressed IH. Caesium (2 mM) depressed the slope of the depolarizing spike pre-potential, resulting in a prolongation of the interspike interval of tonic firing neurons. We conclude that both the inward rectifier current, IH, and the low-threshold calcium current contribute to the spike prepotential of spontaneous action potentials in firing neurons of the rat SCN.

2-Amino-5-phosphonovalerate↗