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1.
J Pharmacol Sci ; 123(3): 219-26, 2013.
Article in English | MEDLINE | ID: mdl-24152964

ABSTRACT

Phosphodiesterase 4 (PDE4) inhibitors have been developed for the treatment of pulmonary inflammatory diseases, but their clinical use was dose-limited by mainly gastric adverse effects. Recent studies suggested PDE4B-selective inhibitors over PDE4D are supposed to display a wider therapeutic index than subtype non-selective PDE4 inhibitors such as roflumilast. Compound A was identified as an orally active PDE4B-selective inhibitor over PDE4D both in humans (80-fold selective) and mice (29-fold selective). In this study, the therapeutic effects of compound A and roflumilast were evaluated on lipopolysaccaride (LPS) injection-induced plasma TNF-α elevation and on LPS inhalation-induced pulmonary neutrophilia in mice. The inhibitory effect on gastric emptying in mice was evaluated as a gastric adverse effect. The therapeutic index for TNF-α production (TI(TNF) = ID50 in gastric emptying / ID50 in LPS injection-induced plasma TNF-α elevation) of compound A was larger than roflumilast (9.0 and 0.2, respectively), whereas the therapeutic index for pulmonary neutrophilia (TI(Neu) = ID50 in gastric emptying / ID50 in LPS inhalation-induced pulmonary neutrophilia) of compound A was comparable to roflumilast (1.0 and 0.5, respectively). In conclusion, the TI(Neu) of compound A was not superior compared to that of roflumilast in spite of its high selectivity for PDE4B over PDE4D in mice.


Subject(s)
Aminopyridines/therapeutic use , Benzamides/therapeutic use , Cyclic Nucleotide Phosphodiesterases, Type 4/metabolism , Heterocyclic Compounds, 2-Ring/therapeutic use , Leukocyte Disorders/drug therapy , Neutrophils , Phenylacetates/therapeutic use , Phosphodiesterase 4 Inhibitors/therapeutic use , Pneumonia/drug therapy , Administration, Ophthalmic , Aminopyridines/administration & dosage , Aminopyridines/adverse effects , Animals , Benzamides/administration & dosage , Benzamides/adverse effects , Cyclopropanes/administration & dosage , Cyclopropanes/adverse effects , Cyclopropanes/therapeutic use , Gastric Emptying/drug effects , Heterocyclic Compounds, 2-Ring/adverse effects , Humans , Leukocyte Disorders/chemically induced , Lipopolysaccharides , Male , Mice , Mice, Inbred BALB C , Neutrophil Infiltration , Phenylacetates/adverse effects , Phosphodiesterase 4 Inhibitors/administration & dosage , Phosphodiesterase 4 Inhibitors/adverse effects , Pneumonia/chemically induced , Tumor Necrosis Factor-alpha/blood
2.
Eur J Pharmacol ; 596(1-3): 153-9, 2008 Oct 31.
Article in English | MEDLINE | ID: mdl-18706408

ABSTRACT

Neurokinins are known to induce neurogenic inflammation related to respiratory diseases. The effects of CS-003 ([1-{2-[(2R)-(3,4-dichlorophenyl)-4-(3,4,5-trimethoxybenzoyl)morpholin-2-yl]ethyl}spiro[benzo[c]thiophene-1(3H),4'-piperidine]-(2S)-oxide hydrochloride]), a novel triple neurokinin receptor antagonist, on several respiratory disease models were evaluated in guinea pigs. As we have already shown that CS-003 is intravenously effective, we first determined if CS-003 was orally effective. CS-003 dose-dependently inhibited substance P-induced tracheal vascular hyperpermeability, neurokinin A- and neurokinin B-induced bronchoconstriction with ID(50) values of 3.6, 1.3 and 0.89 mg/kg (p.o.), respectively. CS-003 (10 mg/kg, p.o.) inhibited the number of coughs induced by capsaicin aerosol (P<0.01) and the antitussive effect was comparable to that of codeine. CS-003 (10 mg/kg, p.o.) also inhibited airway hyperresponsiveness to methacholine chloride in ovalbumin-induced asthma models (P<0.01), a milder one and a severer one. On the other hand, montelukast (10 mg/kg, p.o.), a leukotriene receptor antagonist, significantly inhibited the hyperresponsiveness only in the milder model (P<0.05). In an ovalbumin-induced rhinitis model, oral administration of CS-003 inhibited nasal blockade in a dose-dependent manner and the inhibitory effect was comparable to that of dexamethasone (10 mg/kg, p.o.). CS-003 (i.v.) also dose-dependently inhibited cigarette smoke-induced bronchoconstriction, tracheal vascular hyperpermeability and mucus secretion. These data show that CS-003, a potent orally active triple neurokinin receptor antagonist, may be useful for the treatment of respiratory diseases associated with neurokinins, such as allergic asthma, allergic rhinitis, chronic obstructive pulmonary disease and cough.


Subject(s)
Cyclic S-Oxides/pharmacology , Morpholines/pharmacology , Receptors, Tachykinin/antagonists & inhibitors , Respiratory Tract Diseases/drug therapy , Administration, Oral , Animals , Asthma/drug therapy , Asthma/immunology , Bronchoconstriction/drug effects , Capillary Permeability/drug effects , Capsaicin , Cough/chemically induced , Cough/drug therapy , Cyclic S-Oxides/administration & dosage , Cyclic S-Oxides/therapeutic use , Disease Models, Animal , Guinea Pigs , Male , Morpholines/administration & dosage , Morpholines/therapeutic use , Mucus/metabolism , Ovalbumin , Pulmonary Disease, Chronic Obstructive/drug therapy , Rhinitis/drug therapy , Rhinitis/immunology , Smoke , Nicotiana , Trachea/blood supply , Trachea/metabolism
3.
Eur J Pharmacol ; 586(1-3): 306-12, 2008 May 31.
Article in English | MEDLINE | ID: mdl-18353309

ABSTRACT

Neurokinins are known to induce neurogenic inflammation related to respiratory diseases, though there is little information on triple neurokinin receptor antagonists. The pharmacological properties of the novel triple neurokinin 1, 2 and 3 receptor antagonist [1-(2-[(2R)-(3,4-dichlorophenyl)-4-(3,4,5-trimethoxybenzoyl)morpholin-2-yl]ethyl)spiro[benzo[c]thiophene-1(3H),4'-piperidine]-(2S)-oxide hydrochloride] (CS-003) were evaluated in this study. The binding affinities of CS-003 were evaluated with human and guinea pig neurokinin receptors. As well, the in vivo antagonism of CS-003 against exogenous neurokinins and effects on capsaicin-induced and citric acid-induced responses were investigated in guinea pigs. CS-003 exhibited high affinities for human neurokinin 1, neurokinin 2 and neurokinin 3 receptors with Ki values (mean+/-S.E.M.) of 2.3+/-0.52, 0.54+/-0.11 and 0.74+/-0.17 nM, respectively, and for the guinea pig receptors with Ki values of 5.2+/-1.4, 0.47+/-0.075 and 0.71+/-0.27 nM, respectively. Competitive antagonism was indicated in a Schild analysis of substance P-, neurokinin A- and neurokinin B-induced inositol phosphate formation with pA2 values of 8.7, 9.4 and 9.5, respectively. CS-003 inhibited substance P-induced tracheal vascular hyperpermeability, neurokinin A- and neurokinin B-induced bronchoconstriction with ID50 values of 0.13, 0.040 and 0.063 mg/kg (i.v.), respectively. CS-003 also inhibited capsaicin-induced bronchoconstriction (ID50: 0.27 mg/kg, i.v.), which is caused by endogenous neurokinins. CS-003 significantly inhibited citric acid-induced coughs and the effect was greater than those of other selective neurokinin receptor antagonists. CS-003 is a potent antagonist of triple neurokinin receptors and may achieve the best therapeutic efficacy on respiratory diseases associated with neurokinins compared to selective neurokinin receptor antagonists.


Subject(s)
Cyclic S-Oxides/pharmacology , Morpholines/pharmacology , Neurokinin A/antagonists & inhibitors , Neurokinin A/pharmacology , Neurokinin B/antagonists & inhibitors , Neurokinin B/pharmacology , Neurokinin-1 Receptor Antagonists , Receptors, Neurokinin-2/antagonists & inhibitors , Receptors, Neurokinin-3/antagonists & inhibitors , Animals , Bronchi/drug effects , COS Cells , Capillary Permeability/drug effects , Capsaicin/antagonists & inhibitors , Capsaicin/pharmacology , Chlorocebus aethiops , Citric Acid , Cough/chemically induced , Cough/prevention & control , Dipeptides/pharmacology , Dose-Response Relationship, Drug , Guinea Pigs , Humans , In Vitro Techniques , Indoles/pharmacology , Inositol Phosphates/biosynthesis , Male , Molecular Sequence Data , Receptors, Neurokinin-1/metabolism , Receptors, Neurokinin-2/metabolism , Receptors, Neurokinin-3/metabolism , Substance P/pharmacology , Trachea/blood supply , Trachea/drug effects
4.
J Clin Pharmacol ; 57(9): 1221-1230, 2017 09.
Article in English | MEDLINE | ID: mdl-28464321

ABSTRACT

DS-3801b is an orally active, nonmacrolide, selective motilin receptor agonist. The aim of this 2-part first-in-human study was to assess the safety, tolerability, pharmacokinetics, and pharmacodynamic effects on proximal and distal gastrointestinal (GI) motility of single oral doses of DS-3801b in healthy subjects. The 13 C-octanoate breath test was used to assess gastric emptying (GE), a measure of proximal GI motility. The time to first bowel movement (TTFBM) and the consistency of the first bowel movement according to the Bristol Stool Scale (BSS) were recorded to assess distal GI motility. In part A, 48 subjects received single oral doses of DS-3801b from 1 to 100 mg or placebo (6 DS-3801b, 2 placebo per cohort). In part B, 12 subjects received 50 mg of DS-3801b or placebo to assess GE. DS-3801b is safe and generally well tolerated after doses up to 50 mg, resulting in mild, predominantly GI adverse events. DS-3801a plasma concentrations increase with increasing doses; however, Cmax increases greater than dose-proportionally, whereas AUC increases less than dose-proportionally. The double peaks observed are consistent with multiple absorption sites. Results of the 13 C-octanoate breath test indicate that DS-3801b accelerates GE. Fifty milligrams of DS-3801b resulted in a 20.8% median reduction in GE T1/2 and a 20.6% median reduction in GE Tlag compared with placebo. However, this increase in proximal GI motility was not accompanied by an effect on distal GI motility, as indicated by no significant differences in TTFBM and BSS values across DS-3801b dose levels or compared with placebo.


Subject(s)
Cyclohexanes , Gastrointestinal Motility/drug effects , Piperazines , Receptors, Gastrointestinal Hormone/agonists , Receptors, Neuropeptide/agonists , Adult , Cross-Over Studies , Double-Blind Method , Female , Healthy Volunteers , Humans , Male , Middle Aged , Young Adult
5.
Prog Neurobiol ; 73(1): 1-60, 2004 May.
Article in English | MEDLINE | ID: mdl-15193778

ABSTRACT

This review focuses on the remodeling of brain circuitry associated with epilepsy, particularly in excitatory glutamate and inhibitory GABA systems, including alterations in synaptic efficacy, growth of new connections, and loss of existing connections. From recent studies on the kindling and status epilepticus models, which have been used most extensively to investigate temporal lobe epilepsy, it is now clear that the brain reorganizes itself in response to excess neural activation, such as seizure activity. The contributing factors to this reorganization include activation of glutamate receptors, second messengers, immediate early genes, transcription factors, neurotrophic factors, axon guidance molecules, protein synthesis, neurogenesis, and synaptogenesis. Some of the resulting changes may, in turn, contribute to the permanent alterations in seizure susceptibility. There is increasing evidence that neurogenesis and synaptogenesis can appear not only in the mossy fiber pathway in the hippocampus but also in other limbic structures. Neuronal loss, induced by prolonged seizure activity, may also contribute to circuit restructuring, particularly in the status epilepticus model. However, it is unlikely that any one structure, plastic system, neurotrophin, or downstream effector pathway is uniquely critical for epileptogenesis. The sensitivity of neural systems to the modulation of inhibition makes a disinhibition hypothesis compelling for both the triggering stage of the epileptic response and the long-term changes that promote the epileptic state. Loss of selective types of interneurons, alteration of GABA receptor configuration, and/or decrease in dendritic inhibition could contribute to the development of spontaneous seizures.


Subject(s)
Brain/physiopathology , Disease Models, Animal , Epilepsy, Temporal Lobe/etiology , Epilepsy, Temporal Lobe/physiopathology , Kindling, Neurologic , Neuronal Plasticity , Status Epilepticus/physiopathology , Animals , Brain Mapping , Humans , Neural Inhibition , Neural Pathways/physiopathology
6.
Mutat Res Genet Toxicol Environ Mutagen ; 780-781: 46-50, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25892621

ABSTRACT

As part of a collaborative study by the Mammalian Mutagenicity Study Group of the Environmental Mutagen Society of Japan, we examined micronucleus induction in hepatocytes following oral administration of 2,6-dinitrotoluene (2,6-DNT) at 30, 40, and 50mg/kg/day for 14 days or at 20, 30, and 40mg/kg/day for 28 days to young adult male rats. This compound is known to be a rat liver carcinogen. The formation of micronucleated hepatocytes was confirmed to be dose-dependent with statistically significant increases observed in both treatments. In contrast, no statistically significant changes in the percentage of micronucleated immature erythrocytes were observed in any dose group in the bone marrow micronucleus assay. These results indicated that the repeated-dose liver micronucleus assay has the potential to detect genotoxic hepatocarcinogens and can be integrated into general toxicological studies.


Subject(s)
Carcinogens/toxicity , Dinitrobenzenes/toxicity , Hepatocytes/drug effects , Liver/drug effects , Micronucleus Tests , Administration, Oral , Age Factors , Animals , Body Weight/drug effects , Bone Marrow/drug effects , Chromosome Aberrations/drug effects , Cooperative Behavior , Dose-Response Relationship, Drug , Drug Administration Schedule , Hepatocytes/pathology , Humans , Japan , Liver/pathology , Male , Organ Specificity , Rats , Rats, Sprague-Dawley , Reticulocytes/drug effects , Societies, Pharmaceutical
7.
Neuropsychopharmacology ; 29(7): 1251-8, 2004 Jul.
Article in English | MEDLINE | ID: mdl-15039765

ABSTRACT

There is an increased incidence of schizophrenia-like psychosis in temporal lobe epilepsy (TLE), and several risk factors have been implicated, including the duration of epilepsy and temporal lobe neuropathology. To investigate the biological mechanism of epileptic psychosis, we examined alterations of central dopaminergic systems in the kainate model of TLE. In adult rats, kainate was microinjected into the left amygdala to induce status epilepticus. An indirect dopamine agonist methamphetamine (MAP, 2 mg/kg, i.p.) was administered before and 1 month after the kainate treatment. MAP-induced locomotor activity was significantly enhanced in the kainate group compared with the baseline (pre-kainate) level, which was antagonized by pretreatment with haloperidol. The enhancement of locomotor activity in the kainate group was significantly correlated with the density of hippocampal CA1 neurons. Although the basal extracellular dopamine concentration was significantly lower in the striatum in the kainate group than in the control group (5.5 vs 39.2 fmol/20-min sample), the maximal concentration following MAP administration did not differ between the two groups. These results clearly demonstrate that hypersensitivity of the dopamine systems develops in the chronic phase of the kainate-induced TLE model, which may be responsible for the mechanism of epileptic psychosis.


Subject(s)
Disease Models, Animal , Dopamine/metabolism , Epilepsy, Temporal Lobe/metabolism , Psychotic Disorders/metabolism , Analysis of Variance , Animals , Behavior, Animal , Brain Chemistry/drug effects , Chromatography, High Pressure Liquid/methods , Dopamine Antagonists/pharmacology , Dopamine Uptake Inhibitors/pharmacology , Electrochemistry/methods , Epilepsy, Temporal Lobe/chemically induced , Epilepsy, Temporal Lobe/complications , Haloperidol/pharmacology , In Vitro Techniques , Kainic Acid , Male , Methamphetamine/pharmacology , Microdialysis/methods , Motor Activity/drug effects , Psychotic Disorders/etiology , Rats , Rats, Sprague-Dawley
8.
Neuropsychopharmacology ; 26(6): 794-801, 2002 Jun.
Article in English | MEDLINE | ID: mdl-12007750

ABSTRACT

UNLABELLED: Since delusional disorder is characterized by mono-symptomatic paranoid symptoms, it can be a good clinical model for investigating the dopaminergic mechanism responsible for paranoid symptoms. We examined neuroleptic responses, plasma homovanillic acid (pHVA) and genes of the dopamine receptor (DR) and its synthesizing enzyme (tyrosine hydroxylase: TH) in patients with delusional disorder and compared them with those of schizophrenic patients and healthy controls. RESULTS: (1) A relatively small dose of haloperidol was more effective for delusional disorder than for schizophrenia. (2) The pretreatment level of pHVA was higher in patients with persecution-type, but not in those with jealousy-type delusional disorder, compared with age- and sex-matched controls. This increased pHVA level was decreased eight weeks after successful haloperidol treatment. (3) The genotype frequency of the DRD2 gene Ser311Cys was significantly higher in patients with persecution-type delusional disorder (21%), compared with schizophrenic patients (6%) or controls (6%). (4) Patients homozygous for the DRD3 gene Ser9Ser had higher pretreatment levels of pHVA than those heterozygous for Ser9Gly. (v) A significant positive correlation was found between the polymorphic (TCAT)(n) repeat in the first intron of the TH gene and pretreatment levels of pHVA in delusional disorder. We suggest that delusional disorder, especially the persecution-type, includes a "dopamine psychosis," and that polymorphism of the DRD2, DRD3 and/or TH gene is part of the genetic basis underlying the hyperdopaminergic state that produces paranoid symptoms. Further studies on a large sample size are required.


Subject(s)
Psychotic Disorders/genetics , Receptors, Dopamine/genetics , Schizophrenia, Paranoid/genetics , Adult , Antipsychotic Agents/therapeutic use , Female , Genotype , Homovanillic Acid/blood , Humans , Male , Middle Aged , Psychotic Disorders/blood , Psychotic Disorders/drug therapy , Psychotic Disorders/psychology , Schizophrenia, Paranoid/blood , Schizophrenia, Paranoid/drug therapy , Schizophrenia, Paranoid/psychology
9.
Brain Res ; 1003(1-2): 194-8, 2004 Apr 02.
Article in English | MEDLINE | ID: mdl-15019580

ABSTRACT

Kindling of the ventral tegmental area (VTA), a major source of the mesolimbic dopamine pathway, was examined in rats. We applied two quantitative measurements of dopamine sensitivity before and 2 weeks after VTA kindling (20 times electrical stimulations (100 microA at 1 min intervals) delivered once per day for 14 consecutive days): behavioral responses induced by test VTA stimulation and methamphetamine (MAP)-induced locomotor activity. The total amount of MAP-induced locomotor activity was significantly increased after VTA kindling, while the responses to electrical stimulation were unchanged. These results indicate that repeated activation of the mesolimbic dopamine system can produce a neuroplastic change, which results in dopamine supersensitivity.


Subject(s)
Dopamine/physiology , Kindling, Neurologic/physiology , Ventral Tegmental Area/physiology , Animals , Kindling, Neurologic/drug effects , Male , Methamphetamine/pharmacology , Motor Activity/drug effects , Motor Activity/physiology , Rats , Rats, Sprague-Dawley , Ventral Tegmental Area/drug effects
10.
Eur J Pharmacol ; 498(1-3): 287-94, 2004 Sep 13.
Article in English | MEDLINE | ID: mdl-15364007

ABSTRACT

In this study, we investigated the involvement of neurokinin NK3 receptors in a severe asthma model prepared by administering ovalbumin via inhalation three times to systemically sensitized guinea pigs. [3H]senktide, a neurokinin NK3 receptor ligand, showed significant specific binding to the lungs from the model animals, but not to those from negative control animals. The airway responsiveness to intravenous neurokinin B, a neurokinin NK3 receptor agonist, was increased in the model, indicating an increase in functional NK3 receptors. Furthermore, SB 223956 ((-)-3-methoxy-2-phenyl-N-[(1S)-phenylpropyl]quinoline-4-carboxamide), a selective neurokinin NK3 receptor antagonist, significantly inhibited the ovalbumin-induced airway hyperresponsiveness to inhaled methacholine, but it did not show significant effects on the ovalbumin-induced airway narrowing and eosinophil accumulation. These results suggest that the expressed neurokinin NK3 receptors in the severe asthma model are involved in the development of airway hyperresponsiveness.


Subject(s)
Asthma/metabolism , Receptors, Neurokinin-3/metabolism , Substance P/analogs & derivatives , Animals , Asthma/immunology , Benzamides/metabolism , Binding, Competitive , Bronchoalveolar Lavage Fluid/cytology , Bronchoconstriction/drug effects , Bronchoconstrictor Agents/pharmacology , Disease Models, Animal , Dose-Response Relationship, Drug , Eosinophils/drug effects , Eosinophils/pathology , Guinea Pigs , Lung/drug effects , Lung/pathology , Lung/physiopathology , Male , Methacholine Chloride/pharmacology , Neurokinin B/pharmacology , Ovalbumin/immunology , Peptide Fragments/metabolism , Piperidines/metabolism , Receptors, Neurokinin-3/antagonists & inhibitors , Substance P/metabolism , Tritium
11.
Neurosci Lett ; 340(3): 242-4, 2003 Apr 17.
Article in English | MEDLINE | ID: mdl-12672551

ABSTRACT

We have previously shown that beta-amyloid (Abeta) increased the excitotoxicity of ibotenate, an N-methyl-D-aspartate (NMDA) receptor agonist, to hippocampal neurons of rats. In this report, non-toxic amounts of kainate were co-injected with Abeta into rat hippocampus. Nissl-stained brain sections revealed that Abeta/kainate co-injection exerted synergistic neuronal degeneration in the hippocampus as well as that by Abeta/ibotenate co-injection. MK-801, an NMDA receptor antagonist, blocked the neuronal loss induced by Abeta/ibotenate co-injection, but not by Abeta/kainate co-injection. On the other hand, 6-cyano-7-nitroquinoxaline-2, 3-dione, a kainate receptor antagonist, suppressed the neuronal loss induced by the Abeta/kainate co-injection, but not that by the Abeta/ibotenate co-injection. This suggests that Abeta increases the sensitivity of both the NMDA receptor and the kainate receptor.


Subject(s)
Amyloid beta-Peptides/toxicity , Hippocampus/drug effects , Kainic Acid/toxicity , Peptide Fragments/toxicity , Animals , Drug Synergism , Hippocampus/pathology , Male , Nerve Degeneration/chemically induced , Nerve Degeneration/pathology , Rats , Rats, Sprague-Dawley
12.
Epilepsy Res ; 61(1-3): 113-8, 2004.
Article in English | MEDLINE | ID: mdl-15451013

ABSTRACT

[(123)I]Iomazenil (IMZ) is a specific ligand for central-type benzodiazepine receptors (BZRs) and is available for single photon emission computed tomography (SPECT) to detect epileptogenic foci. We have recently demonstrated time-dependent alterations of [(125)I]IMZ binding in the rat kainate model of temporal lobe epilepsy. Quantitative evaluation of central-type benzodiazepine receptors with [(125)I]Iomazenil in experimental epileptogenesis. I. The rat kainate model of temporal lobe epilepsy. In the present study, we investigated regional changes in central-type BZRs in the cortical dysplasia (CD) model of epilepsy in rats. Pregnant rats were irradiated at day 17 of gestation with 1.2 Gy to produce CD in their pups, and in vitro autoradiography with [(125)I]IMZ was performed at 8 weeks after birth. Intact rats at the same age were used as controls. [(125)I]IMZ binding was significantly decreased in various cortical regions of the in utero irradiated rats, including the bilateral frontal cortex (down to 92-93% of control), cingulate cortex (91-92%), hippocampal areas CA1 (95%), CA2 (94-95%) and CA4 (95-96%), and caudate/putamen (90-94%). In addition, amygdala-kindling was significantly facilitated in the CD model, especially during the late phase of kindling, suggesting seizure susceptibility of this model. These results may replicate the clinical usefulness of central-type BZRs neuroimaging for detection of human epileptogenic CD and indicate dysfunction of GABA-A/BZR-mediated inhibition responsible for the seizure susceptibility.


Subject(s)
Anticonvulsants , Cerebral Cortex/abnormalities , Epilepsy/pathology , Flumazenil/analogs & derivatives , Receptors, GABA-A/drug effects , Abnormalities, Radiation-Induced/pathology , Amygdala/physiology , Animals , Autoradiography , Cerebral Cortex/pathology , Epilepsy/etiology , Female , Hippocampus/pathology , Kindling, Neurologic/physiology , Organ Size , Pregnancy , Rats , Rats, Sprague-Dawley , Receptors, GABA-A/metabolism
13.
Epilepsy Res ; 61(1-3): 105-12, 2004.
Article in English | MEDLINE | ID: mdl-15451012

ABSTRACT

This study aimed at quantitatively evaluating hippocampal central-type benzodiazepine receptors (BZRs) in the kainate model of temporal lobe epilepsy (TLE) by in vitro autoradiography (ARG) using [(125)I] Iomazenil (IMZ) specific ligand for central-type BZRs. Kainate (1 microg/0.5 microl) was injected into the left amygdala to induce limbic status epilepticus. One, three, or six months after injection, in vitro ARG with [(125)I] IMZ and cell counts were performed in the hippocampal CA1-4 regions and dentate gyrus ipsilateral to the kainate injection site, and were compared with the vehicle-injected control group. In all kainate-treated rats, clear pyramidal neuron loss was observed in left hippocampal areas CA1-4. Compared with the control group, progressive reduction of [(125)I] IMZ binding was also observed. This resulted in a marked binding decrease paralleling pyramidal neuron loss in hippocampal areas CA1 (down to 83% of control), CA2 (76%), CA3 (75%), and CA4 (90%) at 6 months after kainate administration. Conversely, [(125)I] IMZ binding significantly increased in the dentate gyrus (up to 106% of control) at 1 month, but returned to nearly normal at 3-6 months. These results suggest that central-type BZR neuroimaging is useful in detecting hippocampal sclerosis in the mesial TLE, though central BZR alterations differ depending on hippocampal subfields and post-seizure time-courses.


Subject(s)
Anticonvulsants , Epilepsy, Temporal Lobe/chemically induced , Epilepsy, Temporal Lobe/metabolism , Excitatory Amino Acid Agonists , Flumazenil/analogs & derivatives , Kainic Acid , Receptors, GABA-A/drug effects , Amygdala , Animals , Autoradiography , Cell Count , Dentate Gyrus/metabolism , Dentate Gyrus/pathology , Electroencephalography/drug effects , Excitatory Amino Acid Agonists/administration & dosage , Hippocampus/metabolism , Hippocampus/pathology , Injections , Iodine Radioisotopes , Kainic Acid/administration & dosage , Ligands , Male , Pyramidal Cells/drug effects , Rats , Rats, Wistar
14.
J Toxicol Sci ; 36(4): 499-505, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21804315

ABSTRACT

The potential genotoxicity of the rodent liver carcinogen 2,6-dinitrotoluene (2,6-DNT) was evaluated in compliance with the guidelines for genotoxicity studies of drugs (Notification No. 1604, Nov. 1, 1999, Ministry of Health and Welfare, Japan) and the OECD guidelines for the testing of chemicals by performing the bacterial reverse mutation (Ames) assay, the in vitro chromosomal aberration assay, and the in vivo comet assay (alkaline single cell gel electrophoresis) in rat liver. In the Ames assay, 2,6-DNT was moderately positive irrespective of metabolic activation. In the in vitro chromosomal aberration assay, under conditions where the test substance would precipitate out, weak structural aberrations were observed with or without S9 mix at each dose at which the cell growth rate was about 40 to 50%. The in vivo comet assay yielded positive results in rat liver; that is to say, the increases in % tail DNA in liver in the 25 and 50 mg/kg groups were observed statistically significantly and dose-dependent. Our findings are in accordance with previous findings in the in vivo/in vitro unscheduled DNA synthesis (UDS) assay in rat liver and in a young rat liver micronucleus assay, although the rat bone marrow micronucleus assay gave negative results. These results suggest that test systems using liver are a useful method for the in vivo genotoxicity assessment of chemicals that require metabolic activation.


Subject(s)
Chromosome Aberrations/chemically induced , DNA Damage , Dinitrobenzenes/toxicity , Mutagenicity Tests/methods , Mutagens/toxicity , Animals , Cell Line , Comet Assay , Cricetinae , Cricetulus , Dose-Response Relationship, Drug , Escherichia coli/genetics , Fibroblasts/drug effects , Fibroblasts/pathology , Liver/drug effects , Liver/pathology , Male , Rats , Rats, Sprague-Dawley , Salmonella typhimurium/genetics
16.
Am J Physiol Lung Cell Mol Physiol ; 296(3): L361-71, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19112102

ABSTRACT

We focused on the regulation of inflammatory mediator expression by adenovirus E1A in lung epithelial cells and the role of this viral protein in the pathogenesis of chronic obstructive pulmonary disease (COPD). We previously reported that E1A, a well-known regulator of host genes, increased ICAM-1 expression in human bronchial epithelial (HBE) and A549 cells in response to LPS stimulation. In this report, we clarified the mechanism of this regulation. We found NF-kappaB translocation to the nucleus after LPS stimulation in both E1A-positive and -negative HBE cells. ICAM-1 promoter reporter constructs revealed that a mutation in the proximal NF-kappaB binding site completely inhibited increased transcription, whereas the mutation in a distal site did not. We analyzed the participation of E1A in transcriptional complex formation at this promoter using chromatin immunoprecipitation. In E1A-positive HBE and A549 cells, LPS stimulation increased ICAM-1 promoter immunoprecipitation by NF-kappaB p65 and p300 but not activator protein-1 antibodies with a concomitant increase by the E1A antibody. No increase was found in E1A-negative cells except in HBE cells with p65 antibody. The association of E1A with the increased promoter immunoprecipitation with p300 was also observed after TNF-alpha stimulation of A549 cells. These results suggest that adenovirus E1A regulates the ICAM-1 promoter through its proximal NF-kappaB binding site, most likely by interacting with the transcriptional complex that forms at this site. E1A regulation of the LPS response may play a role in acute exacerbations as a consequence of bacterial infections in COPD.


Subject(s)
Adenoviridae/genetics , Adenovirus E1A Proteins/genetics , Intercellular Adhesion Molecule-1/genetics , Lung/metabolism , Adenoviridae/pathogenicity , Adenovirus E1A Proteins/physiology , Bacterial Infections/complications , Base Sequence , Binding Sites/genetics , Bronchi/cytology , Bronchi/metabolism , Cell Line , DNA/genetics , DNA/metabolism , Epithelial Cells/metabolism , Genes, Reporter , Host-Pathogen Interactions/genetics , Humans , Inflammation Mediators/metabolism , Lipopolysaccharides/toxicity , Luciferases/genetics , Lung/cytology , Mutation , NF-kappa B/metabolism , Promoter Regions, Genetic , Pulmonary Disease, Chronic Obstructive/etiology , Regulatory Elements, Transcriptional , Toll-Like Receptor 4/genetics , Transcriptional Activation
17.
Am J Physiol Lung Cell Mol Physiol ; 295(1): L79-85, 2008 Jul.
Article in English | MEDLINE | ID: mdl-18469117

ABSTRACT

Epidemiologic and animal studies have shown that exposure to particulate matter air pollution (PM) is a risk factor for the development of atherosclerosis. Whether PM-induced lung and systemic inflammation is involved in this process is not clear. We hypothesized that PM exposure causes lung and systemic inflammation, which in turn leads to vascular endothelial dysfunction, a key step in the initiation and progression of atherosclerosis. New Zealand White rabbits were exposed for 5 days (acute, total dose 8 mg) and 4 wk (chronic, total dose 16 mg) to either PM smaller than 10 mum (PM(10)) or saline intratracheally. Lung inflammation was quantified by morphometry; systemic inflammation was assessed by white blood cell and platelet counts and serum interleukin (IL)-6, nitric oxide, and endothelin levels. Endothelial dysfunction was assessed by vascular response to acetylcholine (ACh) and sodium nitroprusside (SNP). PM(10) exposure increased lung macrophages (P<0.02), macrophages containing particles (P<0.001), and activated macrophages (P<0.006). PM(10) increased serum IL-6 levels in the first 2 wk of exposure (P<0.05) but not in weeks 3 or 4. PM(10) exposure reduced ACh-related relaxation of the carotid artery with both acute and chronic exposure, with no effect on SNP-induced vasodilatation. Serum IL-6 levels correlated with macrophages containing particles (P=0.043) and ACh-induced vasodilatation (P=0.014 at week 1, P=0.021 at week 2). Exposure to PM(10) caused lung and systemic inflammation that were both associated with vascular endothelial dysfunction. This suggests that PM-induced lung and systemic inflammatory responses contribute to the adverse vascular events associated with exposure to air pollution.


Subject(s)
Air Pollutants/toxicity , Endothelium, Vascular/metabolism , Particulate Matter/toxicity , Pneumonia/metabolism , Acetylcholine/pharmacology , Animals , Atherosclerosis/etiology , Atherosclerosis/metabolism , Atherosclerosis/pathology , Atherosclerosis/physiopathology , Endothelins/metabolism , Endothelium, Vascular/pathology , Female , Inflammation/chemically induced , Inflammation/complications , Inflammation/metabolism , Inflammation/physiopathology , Interleukin-6/metabolism , Leukocyte Count , Macrophages, Alveolar/metabolism , Macrophages, Alveolar/pathology , Nitric Oxide/metabolism , Nitroprusside/pharmacology , Platelet Count , Pneumonia/chemically induced , Pneumonia/complications , Pneumonia/pathology , Pneumonia/physiopathology , Rabbits , Vasodilation/drug effects , Vasodilator Agents/pharmacology
18.
Epilepsia ; 46 Suppl 5: 184-8, 2005.
Article in English | MEDLINE | ID: mdl-15987275

ABSTRACT

PURPOSE: Gamma-aminobutyric acid (GABA)-A/benzodiazepine receptors (BZRs) play an important inhibitory role in epileptogenesis. [123I]Iomazenil (123I-IMZ) is a specific ligand for central-type (or neuronal-type) BNRs and is available for single-photon emission computed tomography (SPECT) in brain disorders. We demonstrated alterations of central-type BZRs in human focal epilepsies and their experimental models. METHODS: We examined interictal 123I-IMZ SPECT in patients with mesial temporal lobe epilepsy (MTLE; n = 19) with hippocampal sclerosis and neocortical epilepsy with focal cortical dysplasia (NE-CD; n = 18), and compared those with magnetic resonance imaging (MRI) and 123I-IMP SPECT (for regional cerebral blood flow). We also investigated in vitro autoradiography with (123)I-IMZ at various time courses in the intraamygdala kainate, amygdala kindling, and in-utero irradiation models. RESULTS: In MTLE patients, the epileptogenic hippocampus often showed decreases in both 123I-IMZ and 123I-IMP SPECT. Consistent with those, marked reduction of 125I-IMZ binding was observed in hippocampal CA1-3 regions of the kainate model, which clearly paralleled pyramidal neuronal loss. In contrast, 125I-IMZ binding was increased in the dentate gyrus at 1 month but returned to the normal level at 3-6 months, when frequent spontaneous seizures appeared. The amygdala-kindling model demonstrated similar increases in 125I-IMZ binding in the dentate gyrus without any changes in other brain regions. In NE-CD patients, the epileptogenic foci showed decreased 123I-IMZ binding with relatively normal 123I-IMP SPECT. 125I-IMZ binding also was decreased in the cerebral cortex, hippocampus (areas CA1, 2, and 4), and caudate/putamen of the in-utero irradiation model. CONCLUSIONS: These results indicate that central-type BZRs neuroimaging is useful for detection of epileptogenic foci, but their alterations differ between epilepsy subtypes and time-courses.


Subject(s)
Epilepsies, Partial/physiopathology , Flumazenil/analogs & derivatives , Receptors, GABA-A/physiology , Adult , Amygdala/physiology , Animals , Autoradiography , Cerebral Cortex/abnormalities , Cerebral Cortex/blood supply , Cerebral Cortex/physiopathology , Dentate Gyrus/diagnostic imaging , Dentate Gyrus/embryology , Dentate Gyrus/physiopathology , Disease Models, Animal , Epilepsies, Partial/diagnostic imaging , Epilepsies, Partial/etiology , Epilepsy, Temporal Lobe/diagnostic imaging , Epilepsy, Temporal Lobe/etiology , Epilepsy, Temporal Lobe/physiopathology , Female , Hippocampus/blood supply , Hippocampus/diagnostic imaging , Hippocampus/physiopathology , Humans , Iodine Radioisotopes , Kainic Acid , Kindling, Neurologic/physiology , Magnetic Resonance Imaging , Male , Neocortex/blood supply , Neocortex/physiopathology , Rats , Regional Blood Flow , Tomography, Emission-Computed, Single-Photon/statistics & numerical data
19.
Exp Neurol ; 177(1): 95-104, 2002 Sep.
Article in English | MEDLINE | ID: mdl-12429214

ABSTRACT

Accumulating evidence suggests that inflammation may play an important part in neurodegenerative diseases such as Alzheimer's disease. Inflammation itself, however, is insufficient to produce acute neurodegeneration in vivo. In this report, we determined whether inflammation increases excitotoxicity in hippocampal neurons. A proinflammagen, bacterial endotoxin lipopolysaccharide, was coinjected with ibotenate, an N-methyl-D-aspartate receptor agonist, into rat hippocampus. One week after coinjection, significant neuronal degeneration and severe tissue collapse were observed in the hippocampus. Astroglial and microglial infiltration were also detected. The neurodegeneration was suppressed by dizocilpine maleate, an N-methyl-D-aspartate receptor antagonist. We then examined whether microglial activation takes part in synergistic neuronal loss. One day after the lipopolysaccharide injection into the rat hippocampus, substantial microglial activation and induction of inducible nitric oxide synthase were observed, while neither neuronal nor astrocytic changes were detected. On the other hand, ibotenate injection at the same place 1 day after lipopolysaccharide injection in the hippocampus produced significant neuronal degeneration and gross microglial activation. These results suggest that inflammation by lipopolysaccharide might play an important role in ibotenate/lipopolysaccharide neurotoxicity.


Subject(s)
Excitatory Amino Acid Agonists/toxicity , Hippocampus/pathology , Acute Disease , Animals , Hippocampus/drug effects , Hippocampus/metabolism , Ibotenic Acid/toxicity , Inflammation/chemically induced , Inflammation/metabolism , Inflammation/pathology , Lipopolysaccharides/adverse effects , Male , Microglia/drug effects , Microglia/metabolism , Microglia/pathology , Nerve Degeneration/chemically induced , Nerve Degeneration/metabolism , Nerve Degeneration/pathology , Rats , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate/agonists , Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors , Receptors, N-Methyl-D-Aspartate/physiology
20.
Epilepsia ; 45(12): 1630-5, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15571522

ABSTRACT

PURPOSE: The driving regulations in Japan were amended in 2002, which lifted the absolute ban on driving by persons with epilepsy (PWE) and granted licenses to PWE after a 2-year seizure-free period. METHODS: To survey the effect of the new driving regulations, we sent questionnaires both to the driving authorities (DAs) and to doctors of the Japan Epilepsy Society (JES). RESULTS: Around 1,400 PWE legally obtained a driving license within 1 year after the amendment, licenses were rejected in 157, and 61 had the license withheld for <6 months. In most cases, the attending doctor assessed fitness for driving; 171 doctors responded to the questionnaire. One third of them commented on a positive change in attitude of PWE with respect to driving. Their main remarks included the need to shorten the seizure-free period to qualify for fitness to drive and the need for special guidelines for conditions such as rare seizure occurrence, recently diagnosed epilepsy, or reflex epilepsy. Problems of assessment identified included difficulty in deciding the time for reassessment, distress of PWE over cancellation of license, cost of the assessment, responsibility of the assessing doctors in case of seizure recurrence, and protection of privacy. They requested the DAs to promote publicity about the information and asked the JES to establish a guideline for assessing fitness to drive. CONCLUSIONS: The results highlighted the need for cooperation between the DAs and the JES for further amendment of the regulations as well as the importance of education for the public, patients, and professionals.


Subject(s)
Automobile Driving/legislation & jurisprudence , Epilepsy , Accidents, Traffic/prevention & control , Accidents, Traffic/statistics & numerical data , Attitude of Health Personnel , Attitude to Health , Automobile Driver Examination/legislation & jurisprudence , Automobile Driving/standards , Epilepsy/epidemiology , Government Regulation , Guidelines as Topic/standards , Health Education/methods , Humans , Japan/epidemiology , Patient Education as Topic/legislation & jurisprudence , Physician's Role , Public Opinion , Social Control, Formal , Surveys and Questionnaires , Voluntary Health Agencies/standards
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