Biomedical subjects

Junichi Tanaka

Publications and source records attributed to Junichi Tanaka.

At least 19 recordsLinked to original sources

PubMed2006

Involvement of NMDA receptor mechanisms in the modulation of serotonin release in the lateral parabrachial nucleus in the rat.

Microdialysis was employed to investigate whether N-methyl-d-asparatate (NMDA) glutamate receptor mechanisms are involved in the modulation of serotonin (5-hydoxytryptamine, 5-HT) release in the region of the lateral parabrachial nucleus (LPBN) in freely moving rats. Perfusion of NMDA (10 and 50 microM) through the microdialysis probe significantly enhanced extracellular concentrations of 5-HT and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) in the LPBN area. Local perfusion of the NMDA antagonist dizocilpine (MK801, 10 and 50 microM) did not change the basal 5-HT and 5-HIAA levels in the LPBN area. MK801 (10 microM) administered together with NMDA antagonized the stimulant effect of NMDA (10 microM). The intake of 0.3M NaCl and water induced by subcutaneous injections of the diuretic furosemide (FURO, 10 mg/kg) and the angiotensin converting enzyme inhibitor captopril (CAP, 5 mg/kg) produced significant increases in the 5-HT and 5-HIAA concentrations in the LPBN area. The increased levels of 5-HT and 5-HIAA caused by the combined treatment with FURO and CAP were attenuated by perfusion of MK801 (10 microM). These results indicate the participation of NMDA receptors in the control of 5-HT release in the LPBN area.

Angiotensin-Converting Enzyme Inhibitors↗

PubMed2006

Inhibition of ribosome recruitment induces stress granule formation independently of eukaryotic initiation factor 2alpha phosphorylation.

Cytoplasmic aggregates known as stress granules (SGs) arise as a consequence of cellular stress and contain stalled translation preinitiation complexes. These foci are thought to serve as sites of mRNA storage or triage during the cell stress response. SG formation has been shown to require induction of eukaryotic initiation factor (eIF)2alpha phosphorylation. Herein, we investigate the potential role of other initiation factors in this process and demonstrate that interfering with eIF4A activity, an RNA helicase required for the ribosome recruitment phase of translation initiation, induces SG formation and that this event is not dependent on eIF2alpha phosphorylation. We also show that inhibition of eIF4A activity does not impair the ability of eIF2alpha to be phosphorylated under stress conditions. Furthermore, we observed SG assembly upon inhibition of cap-dependent translation after poliovirus infection. We propose that SG modeling can occur via both eIF2alpha phosphorylation-dependent and -independent pathways that target translation initiation.

Antigens, Surface↗

PubMed2006

Caliciviruses differ in their functional requirements for eIF4F components.

Two classes of viruses, namely members of the Potyviridae and Caliciviridae, use a novel mechanism for the initiation of protein synthesis that involves the interaction of translation initiation factors with a viral protein covalently linked to the viral RNA, known as VPg. The calicivirus VPg proteins can interact directly with the initiation factors eIF4E and eIF3. Translation initiation on feline calicivirus (FCV) RNA requires eIF4E because it is inhibited by recombinant 4E-BP1. However, to date, there have been no functional studies carried out with respect to norovirus translation initiation, because of a lack of a suitable source of VPg-linked viral RNA. We have now used the recently identified murine norovirus (MNV) as a model system for norovirus translation and have extended our previous studies with FCV RNA to examine the role of the other eIF4F components in translation initiation. We now demonstrate that, as with FCV, MNV VPg interacts directly with eIF4E, although, unlike FCV RNA, translation of MNV RNA is not sensitive to 4E-BP1, eIF4E depletion, or foot-and-mouth disease virus Lb protease-mediated cleavage of eIF4G. We also demonstrate that both FCV and MNV RNA translation require the RNA helicase component of the eIF4F complex, namely eIF4A, because translation was sensitive (albeit to different degrees) to a dominant negative form and to a small molecule inhibitor of eIF4A (hippuristanol). These results suggest that calicivirus RNAs differ with respect to their requirements for the components of the eIF4F translation initiation complex.

Animals↗

PubMed2006

Functional characterization of IRESes by an inhibitor of the RNA helicase eIF4A.

RNA helicases are molecular motors that are involved in virtually all aspects of RNA metabolism. Eukaryotic initiation factor (eIF) 4A is the prototypical member of the DEAD-box family of RNA helicases. It is thought to use energy from ATP hydrolysis to unwind mRNA structure and, in conjunction with other translation factors, it prepares mRNA templates for ribosome recruitment during translation initiation. In screening marine extracts for new eukaryotic translation initiation inhibitors, we identified the natural product hippuristanol. We show here that this compound is a selective and potent inhibitor of eIF4A RNA-binding activity that can be used to distinguish between eIF4A-dependent and -independent modes of translation initiation in vitro and in vivo. We also show that poliovirus replication is delayed when infected cells are exposed to hippuristanol. Our study demonstrates the feasibility of selectively targeting members of the DEAD-box helicase family with small-molecule inhibitors.

Adenosine Triphosphate↗

PubMed2006

Non-NMDA glutamatergic receptors modulate acetylcholine release in the rat subfornical organ area.

The present study was designed to examine whether glutamatergic receptor mechanisms modulate the release of acetylcholine (ACh) in the region of the subfornical organ (SFO) using intracerebral microdialysis methods in freely moving rats. Perfusion of either non-N-methyl-d-aspartate (NMDA) agonist quisqualic acid (QA, 50 microM) or kainic acid (KA, 50 microM) through the microdialysis probe significantly enhanced the ACh release in the SFO area. Local perfusion of the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 and 50 microM) did not change the basal release of ACh. CNQX (10 microM) administered together with either QA (50 microM) or KA (50 microM) in the SFO area antagonized the stimulant effect of the agonists on the ACh release. In urethane-anesthetized rats, repetitive electrical stimulation (500 microA, 10 Hz) of the medial septum (MS) significantly increased dialysate ACh concentrations in the region of the SFO. The increase in the ACh release elicited by the MS stimulation was significantly potentiated by perfusion of QA (50 microM), and the QA-induced potentiation was prevented by CNQX (10 microM) treated together with QA. These results show that the glutamatergic synaptic inputs enhance the ACh release in the SFO area through non-NMDA receptors. The data further suggest that the septal cholinergic inputs to the SFO area are potentiated by non-NMDA receptor mechanisms.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed2006

Real-time in vivo virtual histology of colorectal lesions when using the endocytoscopy system.

BACKGROUND: The histological findings of GI lesions are based on light-microscopic examination of H&E-stained thin-slice specimens. Recently, a concept of optical biopsy has been advocated. A study of the observation of colorectal lesions using endocytoscopy to obtain real-time histological images in vivo during endoscopy was performed. DESIGN: Prospective study. AIM: To evaluate the usefulness of optical biopsy of colorectal lesions with the endocytoscopy (E-C) system. PATIENTS: The subjects were 113 consecutive patients who underwent a complete colonic examination, from April 2003 to March 2004, performed by a single colonoscopist. SETTING: Digestive Disease Center of Showa University Northern Yokohama Hospital. RESULTS: With E-C, it was possible to observe lesions at the cellular level and evaluate cellular atypia in addition to structural atypia in vivo. The correlation was statistically significant between the endocytoscopic diagnosis and the histological diagnosis. LIMITATIONS: The endocytoscope had to be touched to the target colonic glands. CONCLUSIONS: It was possible to distinguish neoplastic from non-neoplastic lesions, and also possible to distinguish invasive cancer from adenoma. "Ultra-high" magnifying endoscopy, the E-C system, provides real-time histological images in vivo, which correspond well with those of H&E-stained microscopic images.

Adenoma↗

PubMed2006

Hepatitis C virus-related internal ribosome entry sites are found in multiple genera of the family Picornaviridae.

The internal ribosome entry site (IRES) elements from porcine enterovirus 8 and simian virus 2, two members of a proposed new genus within the family Picornaviridae, were characterized. These IRES elements, in common with the porcine teschovirus 1 IRES, were found to be related functionally and structurally to the IRES element from Hepatitis C virus, a member of the family Flaviviridae. Partial secondary structure predictions were derived and functional assays demonstrated that these IRES elements continued to be active when eIF4G was cleaved and when the activity of eIF4A was blocked.

5' Untranslated Regions↗

PubMed2005

Structural basis of swinholide A binding to actin.

Marine toxins targeting the actin cytoskeleton represent a new and promising class of anti-cancer compounds. Here we present a 2.0 A resolution structure of swinholide A, a marine macrolide, bound to two actin molecules. The structure demonstrates that the actin dimer in the complex does not represent a physiologically relevant entity, for the two actin molecules do not interact with each other. The swinholide A actin binding site is the same as that targeted by toxins of the trisoxazole family and numerous actin binding proteins, highlighting the importance of this site in actin polymerization. The observed structure reveals the mechanism of action of swinholide A and provides a structural framework about which to design new agents directed at the cytoskeleton.

Actins↗

PubMed2005

[A trial of TS-1 administration on the basis of the pharmacokinetic study for an advanced gastric cancer patient with impaired renal function].

TS-1 is an antitumor drug including 5-chloro-2,4 dihydroxypyridine (CDHP), which inhibits dihydriopyrimidine dehydrogenase (DPD) activity selectively in metabolism of 5-FU. However, TS-1 therapy tends to increase adverse events for patients with impaired renal function due to excessively high blood concentration of 5-FU, because CDHP is mainly excreted into the urine. In a 67-year-old male with advanced gastric cancer, renal dysfunction occurred during TS-1 administration as its adverse event. We studied the pharmacokinetics of 5-FU, which were analyzed on the T1/2 value and the AUC (0-infinity) of 5-FU with a single and consecutive TS-1 administration, and estimated an optimal TS-1 administration regimen for this patient. The regimen is 60 mg/body/day given in one divided dose for 28 days consecutively followed by 14 days rest. This regimen enabled a continuation of TS-1 treatment for the patient. In conclusion, individual dose adjustment using pharmacokinetic study of 5-FU might be beneficial to patients with impaired renal function.

Adenocarcinoma↗

PubMed2005

[A case of peritoneal dissemination disappeared by CPT-11 + TS-1 combination chemotherapy].

A patient is a 35-year-old man. By a diagnosis of descending colon cancer, descending colon ablative operation and D1 lymph node dissection were performed on April 22, 2004. It was P3H0N1SE, Stage IV in perioperative findings. Abdominal CT showed peritoneal dissemination of 1.7 cm at the right under the abdominal wall wound and 1.2 cm in the rectovesical pouch on May 18, 2004. CPT-11 + TS-1 combination chemotherapy was started on June 22nd. In the five weeks of the combination chemotherapy, continuous infusion of CPT-11 (150 mg/body day 1 and 15) was twice administered, and oral administration of TS-1 (120 mg/body/day) was given for 3 weeks (day 1-21). Peritoneal dissemination disappeared after the two-course end, and we judged it as CR. Furthermore, we were certain that we obtained CR after the three course end. The adverse event was only neutropenia of grade 1. The fourth course was not administered, but recurrence has not been observed. Abdominal CT showed no recurrence on March 3, 2005 since the combination chemotherapy ended 6 months ago.

Administration, Oral↗

PubMed2004

Extracellular level of basolateral amygdalar dopamine responding to reversal of appetitive-conditioned discrimination in young and old rats.

Young and old rats, aged 3 and 24 months old, respectively, were conditioned to press a lever under lamp-on conditions for reward acquisition and lamp-off for no reward using a variable interval reinforcement schedule that averaged 15 s; i.e., the minimal requirement was 4 responses/min. Over a 30-day period, young and old groups showed increased response to lamp-on from 22 to 35/min and from 10 to 23/min, respectively, and shortened response to lamp-off after initial training. Response to lamp-on as a percentage of total response to lamp-on and -off (the discrimination ratio) was over 80%. For the next 30 days, reversal learning was imposed to reinforce discrimination of the lamp-off state. Young rats showed a steadily increasing discrimination ratio from 40% to 70%, and old rats from 30% to 60%. In comparison with the initial training, young rats showed a total response increase from 50% to 60%, while old rats showed an approximately 5% decrease without loss of reward-obtaining efficiency. In vivo microdialysis during reversal revealed that young rats had higher dopamine transmission in the basolateral amygdala than old rats. The dopamine level was positively correlated with the number of responses to state of reward in young rats and negatively with the numbers of both NRF and RF responses to lamp-on and -off states in old rats. These results suggest that in reversal discrimination, basolateral amygdalar dopamine efflux correlates with the manner of age-related conditioned response rather than the ability to learn.

Age Factors↗

PubMed2004

GABA-mediated attenuation of noradrenaline release in the rat median preoptic area caused by intravenous injection of metaraminol.

Previous studies have shown that the noradrenergic system in the median preoptic nucleus (MnPO) play an important role in the control of the body fluid balance and cardiovascular function and that the release of noradrenaline in the MnPO is regulated by gamma-aminobutyric acid (GABA) receptor mechanisms. The present study was carried out to examine whether the GABAergic system is involved in the modulation of the noradrenaline release in the MnPO in response to an elevation in blood pressure using in vivo microdialysis techniques. In urethane-anaesthetised male rats, the rise in arterial pressure caused by intravenous administration of the alpha-agonist metaraminol significantly decreased dialysate noradrenaline concentration in the MnPO area. The decrease in the noradrenaline level elicited by the metaraminol administration was significantly attenuated by perfusion with either bicuculline (10 microM), a GABA(A) receptor antagonist, or phaclofen (10 microM), a GABA(B) receptor antagonist, through a microdialysis probe. The amount of the antagonist-induced attenuation was much greater in the bicuculline-treated group than in the phaclofen-treated group. These results suggest that the release of noradrenaline in the MnPO area may be modulated by neural inputs from the peripheral baroreceptors, and that the neural inputs may be mediated in part through GABA(A) receptors rather than GABA(B) receptors in the MnPO area.

Adrenergic alpha-Agonists↗

PubMed2004

Involvement of serotonergic systems in the lateral parabrachial nucleus in sodium and water intake: a microdialysis study in the rat.

The present study was carried out to investigate whether 0.3 M NaCl and water intake alters the release of serotonin (5-hydoxytryptamine, 5-HT) in the region of the lateral parabrachial nucleus (LPBN) in freely moving rats. The ingestion of 0.3 M NaCl and water was induced by subcutaneous injections of the diuretic furosemide (FURO, 10 mg/kg) and the angiotensin converting enzyme inhibitor captopril (CAP, 5 mg/kg), and extracellular concentrations of 5-HT and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) were measured using intracerebral microdialysis techniques. The intake of 0.3 M NaCl and water significantly enhanced the 5-HT and 5-HIAA levels in the LPBN area. The combined treatment with FURO and CAP elicited significant decreases in the 5-HT and 5-HIAA concentrations in the LPBN area under the condition that 0.3 M NaCl and water are not available for drinking. These results suggest that the serotonergic system in the LPBN area may play an important role in the modulation of sodium appetite and thirst.

Angiotensin-Converting Enzyme Inhibitors↗

PubMed2004

Absolute stereochemistry of ulapualide A.

[structure: see text] The structure of ulapualide A (1) has been solved by X-ray crystallography in a complex with G-actin. The stereochemical configuration was assigned as 3S,9S,22S,23R,24S,26S,27S,31R,32R,33R.

Actins↗

PubMed2004

Polyamine levels in brain and plasma after acute restraint or water-immersion restraint stress in mice.

To investigate the relationship between polyamines and stress, we measured polyamine levels in the frontal cortex, hippocampus, hypothalamus, and plasma of mice after acute restraint or water-immersion restraint stress. In all parts of the brain, putrescine levels were elevated (139-157% of the control) 24 h after water-immersion restraint stress. In the case of restraint, however, elevation of the putrescine level (130% of the control) was detected only in the frontal cortex. Spermidine and spermine levels were unchanged or slightly reduced (80-85% of the control) in the brain 6 and 24 h after water-immersion restraint stress. There was no change in plasma polyamine levels at any time subsequent to the stress. Pretreatment with diazepam (5 mg/kg, i.p.) completely blocked the stress-induced putrescine increases. These results indicate that the magnitude of the putrescine increase is dependent upon the intensity of the stressor, and suggest that polyamine metabolism is linked to psychological stress.

Animals↗

PubMed2004

Modulation of noradrenaline release in the median preoptic area by GABAergic inputs from the organum vasculosum of the lamina terminalis in the rat.

Previous observations have shown that gamma-aminobutyric acid (GABA) receptor mechanisms modulate the release of noradrenaline (NA) in the median peptic nucleus (MnPO). The present study was carried out to investigate whether neural inputs from the organum vasculosum of the lamina terminalis (OVLT) to the MnPO are involved in the GABAergic modulation of NA release in the MnPO area using in vivo microdialysis techniques. In urethane-anesthetized rats, electrical stimulation (5 and 10 microA, 10Hz) of the OVLT region, but not its surrounding region, significantly enhanced dialysate NA concentration in the MnPO area. The enhancement in the NA level caused by the OVLT region stimulation was significantly increased by perfusion with either bicuculline (10 microM), a GABA(A) receptor antagonist, or phaclofen (10 microM), a GABA(B) receptor antagonist, through a microdialysis probe. The amount of the antagonist-induced increase was much greater in the phaclofen-treated group than in the bicuculline-treated group. These results show that the OVLT region may exert both excitatory and inhibitory influences on the release of NA in the MnPO area, and imply that the inhibitory influence may be mediated through GABA(B) receptors rather than GABA(A) receptors.

Animals↗

PubMed2004

Briarane diterpenes from two species of octocorals, Ellisella sp. and Pteroeides sp.

Eight new briarane diterpenes (1-4, 7-10) have been isolated from two species of octocorals and the structures elucidated by spectroscopic analysis. Two diterpenes (2, 3) from the gorgonian Ellisella sp. inhibited cytokinesis, causing multinuclei formation on NBT-II cells, while a known briarane (12) from the sea pen Pteroeides sp. showed reversal of multidrug resistance.

Animals↗

PubMed2003

Biomolecular mimicry in the actin cytoskeleton: mechanisms underlying the cytotoxicity of kabiramide C and related macrolides.

This study characterizes the interactions between kabiramide C (KabC) and related macrolides and actin and establishes the mechanisms that underlie their inhibition of actin filament dynamics and cytotoxicity. The G-actin-KabC complex is formed through a two-step binding reaction and is extremely stable and long-lived. Competition-binding studies show that KabC binds to the same site on G-actin as Gelsolin domain 1 and CapG. KabC also binds to protomers within F-actin and results in the severing and capping of the (+) end; these studies suggest that free KabC and related macrolides act as biomimetics of Gelsolin. The G-actin-KabC complex binds to the (+) end of a growing filament, where it functions as a novel, unregulated, (+)-end capper and is largely responsible for the inhibition of motility and cytokinesis in approximately 10 -100 nM KabC-treated cells. KabC and related macrolides are useful probes to study the regulation of the actin filament (+) end and may lead to new therapies to treat diseases of the actin cytoskeleton.

2-Naphthylamine↗