Biomedical subjects

Hideo Inoue

Publications and source records attributed to Hideo Inoue.

17 recordsLinked to original sources

PubMed2006

Up-regulation of protease-activated receptor-2 by bFGF in cultured human synovial fibroblasts.

Protease-activated receptors (PARs) have been implicated in the development of acute and chronic inflammatory responses. We have examined the expression of mRNA for PARs and their regulation by growth factors and cytokines in synovial fibroblasts derived from patients with rheumatoid arthritis (RA). Messenger RNA for PAR-1, -2 and -3 was detected in these cells, but not that for PAR-4. Expression of mRNA for PAR-2 was up-regulated by bFGF in a concentration-dependent manner, whereas expression of mRNA for PAR-1 and PAR-3 was not affected. Levels of mRNA encoding PAR-1, PAR-2 and PAR-3 did not increase in response to IL-1beta and TNF-alpha. Expression of mRNA for PAR-2 was maximal 12 h after addition of bFGF, and maximal levels of immunoreactive PAR-2 were reached after 24 h. Furthermore, PAR-2 agonist peptide (SLIGKV-NH(2)), but not the inactive reverse peptide (VKGILS-NH(2)), induced transitory cytosolic Ca(2+) mobilization in cells, and its response was increased by pretreatment with bFGF. An important role could be played by bFGF in the regulation of functional PAR-2 expression in cultured RA synovial fibroblasts.

Calcium↗

PubMed2006

Constitutive expression of thrombospondin 1 in MC3T3-E1 osteoblastic cells inhibits mineralization.

Thrombospondin 1 (TSP1) is a multifunctional extracellular glycoprotein present mainly in the fetal and adult skeleton. Although an inhibitory effect of TSP1 against pathological mineralization in cultured vascular pericytes has been shown, its involvement in physiological mineralization by osteoblasts is still unknown. To determine the role of TSP1 in biomineralization, mouse osteoblastic MC3T3-E1 cells were cultured in the presence of antisense phosphorothioate oligodeoxynucleotides complementary to the TSP1 sequence. The 18- and 24-mer antisense oligonucleotides caused concentration-dependent increases in the number of mineralized nodules, acid-soluble calcium deposition in the cell/matrix layer, and alkaline phosphatase activity within 9 days, without affecting cell proliferation. The corresponding sense or scrambled oligonucleotides did not affect these parameters. In the antisense oligonucleotide-treated MC3T3-E1 cells, thickened extracellular matrix, well-developed cell processes, increased intracellular organelles, and collagen fibril bundles were observed. On the other hand, the addition of TSP1 to the culture decreased the production of a mineralized matrix by MC3T3-E1 cells. Furthermore, MC3T3-E1 clones overexpressing mouse TSP1 were established and assayed for TSP1 protein and their capacity to mineralize. TSP1 dose-dependently inhibited mineralization by these cells both in vitro and in vivo. These results indicate that TSP1 functions as an inhibitory regulator of bone mineralization and matrix production by osteoblasts to sustain bone homeostasis.

Alkaline Phosphatase↗

PubMed2006

Synthesis and ribonuclease activity of oligonucleotides with N3-terpyridine x Cu(II)-linked thymine residues.

In previous studies of RNA cleavage, we prepared 2'-O-methyloligonucleotide-terpyridine x Cu(II) complex(es) as sequence-specific artificial ribonucleases, in which the terpyridine group was attached to the nucleoside-sugar moiety. We found that the antisense 2'-O-methyloligomer with two terpyridine x Cu(II) complexes at contiguous internal sites was most active, and precisely cleaved the target RNA oligomer at the predetermined site. In order to explore more efficient cleavers, we have constructed a structurally similar 18-mer, but with two modified thymidine residues, in which the complex was attached to the base-N(3) position by a hydroxyethyl linker. The reaction of the target RNA 24-mer with a 10-fold excess of the new agent, in the presence of Cu(II) ions, and at pH 7.5 and 37 degrees C, revealed that the substrate was almost completely cleaved after 10 hr, and that the cleavage occurred at two contiguous sites. The activity of the new cleaver was more than half of that of the previous cleaver.

Oligonucleotides↗

PubMed2005

Inhibition by licochalcone A, a novel flavonoid isolated from liquorice root, of IL-1beta-induced PGE2 production in human skin fibroblasts.

Licochalcone A, a novel flavonoid isolated from the root of Glycyrrhiza inflata, has been reported to exhibit anti-inflammatory activity in animal models. In this study, we examined the effect of licochalcone A on the production of chemical mediators such as prostaglandin (PG)E2 and cytokines by interleukin (IL)-1beta in human skin fibroblasts. Licochalcone A (IC50 15.0 nM) inhibited PGE2 production, but not IL-6 and IL-8 production, in response to IL-1beta. NS-398 (IC50 1.6 nM), a COX-2 selective inhibitor, also suppressed the PGE2 production. Furthermore, licochalcone A and NS-398 suppressed PGF(2alpha) production by IL-1beta. However, licochalcone A (1 microM) had no effect on increased levels of cyclooxygenase (COX)-2 mRNA and protein in cells. Dexamethasone (100 nM) not only inhibited PGE2, PGF(2alpha), IL-6 and IL-8 production but also strongly suppressed the expression of COX-2 mRNA and protein. Licochalcone A had no effect on COX-1-dependent PGE2 production, whereas indometacin (100 nM), a dual inhibitor of COX-1 and COX-2, was very effective. These results suggest that licochalcone A induces an anti-inflammatory effect through the inhibition of COX-2-dependent PGE2 production. Furthermore, it appears that the inhibitory effect of licochalcone A on PGE2 production in response to IL-1beta is quite different from that of the steroid.

Base Sequence↗

PubMed2005

Synthesis and inhibitory effect of novel glycyrrhetinic acid derivatives on IL-1 beta-induced prostaglandin E(2) production in normal human dermal fibroblasts.

Olean-11,13(18)-dien-3beta,30-diol dihemiphthalate (3), which was derived from glycyrrhetinic acid (GA), has been reported to produce a potent of anti-inflammatory effect in in vivo assays. Using 3 as a lead compound, we attempted to synthesize some modified compounds which varied in the following; i) the position of a carboxyl group in the phthalate moiety, ii) the number of carboxyls attached to the benzoyl group, iii) conversion of benzene ring to another ring system, iv) the linkage form between the benzene ring and oleanene skeleton at position 3 and/or 30. These were screened for their inhibitory activity against interleukin-1 beta (IL-1 beta)-induced prostaglandin E(2) (PGE(2)) production in normal human dermal fibroblasts (NHDF). Although conversion of the ortho-carboxyl group of 3 into the meta-position or the para-position led to an increase in inhibitory activity, the elimination or increase of the carboxyl group resulted in loss of the inhibitory activity. Conversion of the ester bond to the amide bond at position 3 and/or 30 of 3 did not contribute to a significant increase in inhibitory activity. On the other hand, among the derivatives possessing an anthranilic acid moiety at position 30 of 3beta-O-acetyl-olean-11,13(18)-dien-30-oic acid (20), 3beta-hydroxy-30-nor-olean-11,13(18)-dien-20 beta-[N-(2-carboxyphenyl)]carboxamide (30) showed the most potent inhibitory activity (IC(50) 1.0 microM) in this series.

Cells, Cultured↗

PubMed2004

Guillain-Barré syndrome and hemophagocytic lymphohistiocytosis in a patient with severe chronic active Epstein-Barr virus infection syndrome.

Epstein-Barr virus (EBV) infection causes a wide range of neurologic and hematologic manifestations. We report a 72-year-old Japanese male patient with severe chronic active EBV infection syndrome (SCAEBV) who presented with Guillain-Barré syndrome (GBS) and developed hemophagocytic lymphohistiocytosis (HLH) several months after the onset of GBS. He showed acute onset of distal muscle weakness, ophthalmoplegia and bulbar palsy. Results of nerve conduction study revealed acute motor-sensory axonal neuropathy (AMSAN). His serum was positive for anti-LM1 IgG and anti-GM1b IgM. Titers of antibodies to EBV-related antigens indicated chronic reactivated EBV infection. Treatment with IVIg resolved the acute ophthalmoplegia, but there was no notable improvement in the AMSAN and bulbar palsy despite repeated. Finally, he developed refractory HLH resulting in a fatal outcome. In the present patient, it seems that SCAEBV was associated with the development of GBS and fatal HLH via parainfectious autoimmunity and direct infectious immune mechanisms, respectively.

Aged↗

PubMed2004

Neuro-Behçet's disease with chorea after remission of intestinal Behçet's disease.

Neuro-Behçet's disease shows various neuropsychiatric symptoms, but chorea has rarely been reported. We report a case of neuro-Behçet's disease in a 67-year-old woman with depression and chorea that occurred 22 years after the onset of intestinal Behçet's disease. Brain magnetic resonance imaging (MRI) using a fluid-attenuated inversion-recovery (FLAIR) sequence demonstrated lesions more clearly than did T2-weighted MRI. Some of the lesions appeared as small ring-like foci, i.e. low-intensity spots rimmed with remarkable hyperintense signals, in the periventricular white matter and basal ganglia. A review of the literature revealed that the onset of chorea in cases of Behçet's disease varied from the time of onset of Behçet's disease to 31 years after onset of the disease. Psychiatric manifestations have often been associated with neuro-Behçet's disease. In the present patient, treatment with prednisolone resolved the chorea, suggesting that the chorea was caused by an autoimmune mechanism. It seems likely that the long-term development of vasculitis in patients with Behçet's disease results in the formation of these particular brain lesions on FLAIR MR images. Chorea should be taken into consideration as one of the manifestations of Behçet's disease, even many years after remission of the disease.

Aged↗

PubMed2004

Cloning and expression of the enolase gene from Desulfovibrio vulgaris (Miyazaki F).

The gene encoding an enolase from Desulfovibrio vulgaris (Miyazaki F) was cloned and overexpressed in Escherichia coli. A 2.1-kb DNA fragment, isolated from D. vulgaris (Miyazaki F) by double digestion with PstI and BamHI, contained an enolase gene (eno) and part of the methylenetetrahydrofolate dehydrogenase gene (folD). The nucleotide sequence of eno indicates that the protein monomer is composed of 434 amino acids. An expression system for eno under control of the T7 promoter was constructed in E. coli. The purified His-tagged enolase formed a homooctamer and was active in the formation of phosphoenolpyruvate (PEP) as well as in the reverse reaction, the formation of D-(+)-2-phosphoglyceric acid (2-PGA). The pH dependence and kinetic properties of the recombinant enolase from the sulfate-reducing bacterium were also studied. The amounts of eno mRNA when the bacterium was grown on glycerol or glucose were compared to that when D. vulgaris was grown on lactate.

Amino Acid Sequence↗

PubMed2004

Synthesis and hybridization studies on oligonucleotide-metal complex conjugates.

We synthesized a DNA oligomer with a terpyridine x Cu(II) complex-linked thymidine residue at phosphors the 5'-end. The attachment of the 5'-complex slightly enhanced the thermal stability of the DNA match-duplex. We also determined the thermal stabilities of the modified DNA duplexes and their unmodified counterparts, with a mismatched base-pair, to examine the effect of the complex on the stability.

Base Pair Mismatch↗

PubMed2003

Highly efficient catalytic RNA cleavage by the cooperative action of two Cu(II) complexes embodied within an antisense oligonucleotide.

Based on our recent studies of RNA cleavage by oligonucleotide-terpyridine.Cu(II) complex 5'- and/or 3'-conjugates, we designed 2'-O-methyloligonucleotides with two terpyridine-attached nucleosides at contiguous internal sites. To connect the 2'-terpyridine-modified uridine residue at the 5'-side to the 5'-O-terpyridyl nucleoside residue at the 3'-side, a dimethoxytrityl derivative of 5-hydroxypropyl-5'-O-terpyridyl-2'-deoxyuridine-3'-phosphoramidite was newly synthesized. Using this unit, we constructed two terpyridine conjugates, with either an unusual phophodiester bond or the bond extended by a propanediol(s)-containing linker. Cleavage reactions of the target RNA oligomer, under the conditions of conjugate excess in the presence of Cu(II), indicated that the conjugates precisely cleaved the RNA at the predetermined site and that one propanediol-containing linker was the most appropriate for inducing high cleavage activity. Furthermore, a comparison of the activity of the propanediol agent with those of the control conjugates with one complex confirmed that the two complexes are required for efficient RNA cleavage. The reaction of the novel cleaver revealed a bell-shaped pH-rate profile with a maximum at pH approximately 7.5, which is a result of the cooperative action of the complexes. In addition, we demonstrated that the agent catalytically cleaves an excess of the RNA, with the kinetic parameter kcat/K(m) = 0.118 nM(-1) x h(-1).

Base Sequence↗

PubMed2003

Cloning and expression of the superoxide dismutase gene from the obligate anaerobic bacterium Desulfovibrio vulgaris (Miyazaki F).

We identified the SOD gene in the obligate anaerobic bacterium Desulfovibrio vulgaris (Miyazaki F) and constructed a high-level expression system in Escherichia coli. A 2.6-kbp DNA fragment isolated from D. vulgaris (Miyazaki F) by double digestion with EcoRI and SmaI contained the SOD gene and part of another open reading frame. The amino acid sequence deduced from the SOD gene, which was composed of 238 amino acid residues, showed high homogeneity with iron-containing SOD (Fe-SOD) and predicted that the amino terminus of this protein would carry an export signal peptide. We produced the precursor form of SOD (PSOD) and the mature form of SOD (MSOD), which lacked the putative signal peptide. In E. coli, PSOD was present in insoluble inclusion bodies, and its putative signal peptide was not cleaved. In contrast, MSOD contained one iron per mononer and formed a dimer, which exhibited an SOD activity of 850 U/mg. Furthermore, D. vulgaris soluble extract showed a band of SOD activity on native polyacrylamide gel that migrated to the same point as MSOD. The intracellular localization of SOD and its role in D. vulgaris are also discussed.

Amino Acid Sequence↗

PubMed2003

Oligonucleotide duplex stabilization by end-linked terpyridine x Cu(II) complexes.

As a part of our recent studies of RNA cleavage using antisense oligonucleotide-metal complex(es) conjugates, we prepared self-complementary 2'-O-methyloligonucleotides with a terpyridine x Cu(II) complex, which was attached to the sugar portion of the 5'-end or 3'-end. The thermal stabilities of the formed duplexes, as compared with those of the unmodified 2'-O-methyl duplexes, revealed that both the 5'- and 3'-complexes greatly enhanced the duplex stability.

Base Sequence↗

PubMed2003

Effects of glycyrrhetinic acid derivatives on hepatic and renal 11beta-hydroxysteroid dehydrogenase activities in rats.

The purpose of this study was to examine the structure and activity relationships of glycyrrhetinic acid derivatives on the inhibition of hepatic and renal 11beta-hydroxysteroid dehydrogenases (HSDs) in rats. Furthermore, we explored whether inflammatory effect of the derivatives is involved in the inhibition of 11beta-HSD activity. 18beta-Glycyrrhetinic acid (Ia) potently inhibited 11beta-HSD activity of hepatic (IC50 (concentration giving 50% inhibition of cortisone production) = 0.09 microM) and renal (IC50 = 0.36 microM) homogenate. The inhibitory effect of 18beta-glycyrrhetol (Id) modified at the 30-position of glycyrrhetinic acid was weaker than that of glycyrrhetinic acid itself. 18beta-24-Hydroxyglycyrrhetinic acid (Ie), oxidized at the 24-position, remarkably reduced the inhibitory activity for both enzymes. 18beta-11-Deoxoglycyrrhetinic acid (IIc) showed the same inhibitory effect as glycyrrhetinic acid on hepatic 11beta-HSD activity, but less effect on renal 11beta-HSD activity. Furthermore, the inhibitory activity of 18beta-deoxoglycyrrhetol (IIa), modified at the 11- and 30-position, was markedly decreased. Dihemiphthalate derivatives (IIb, IIIb and IVb) of deoxoglycyrrhetol (IIa), 18beta-olean-9(11), 12-diene-3beta, 30-diol (IIIa) and olean-11, 13(18)-diene-3beta, 30-diol (IVa), which are anti-inflammatory agents, also showed weak inhibition against both hepatic and renal 11beta-HSDs. While glycyrrhetinic acid (200 mg kg(-1), p.o.) significantly inhibited 11beta-HSD activity in rat liver and kidney at 3 h after administration, compound IVb (100 mg kg(-1), p.o.) had no effect on either enzyme activity. In addition, the circulating corticosterone level was slightly increased by glycyrrhetinic acid but not by compound IVb. These results suggest that the anti-inflammatory effects of compound IVb, derived from glycyrrhetinic acid, are not due to accumulation of steroids induced by the inhibition of 11beta-HSD activity. Our data also showed that the 11-, 24- and 30-positions of glycyrrhetinic acid may play important roles in the differential inhibitory effects on 11beta-HSD isozyme activity.

11-beta-Hydroxysteroid Dehydrogenases↗

PubMed2002

Synthesis, characterization and platelet adhesion of segmented polyurethanes grafted phospholipid analogous vinyl monomer on surface.

New segmented polyurethanes (SPUs) grafted phospholipid analogous vinyl monomer, 2-(methacryloyloxy)ethyl phosphorylcholine (MPC) on surface were synthesized. The soft segment of these polyurethanes was hydroxylated poly(isoprene) diol and the hard segments were 4,4'-methylenediphenyl diisocyanate (MDI) and 1,4-butanediol (BD). SPUs were hydroxylated by potassium peroxodisulfate and MPC was grafted on the surface of hydroxylated SPUs using di-ammonium cerium (IV) nitrate (ceric ammonium nitrate, CAN) as a radical initiator. The bulk characterization of synthesized SPUs was investigated by infrared spectroscopy (IR) and gel-permeation chromatography (GPC). The existence of phospholipid analogous groups on the surface of these SPUs was revealed by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS) and contact angle measurements. The surfaces of MPC-grafted SPUs showed decreased water contact angles compared to non-grafted SPU and the presence of phosphorylcholine groups. The blood compatibilities of the new polymers were evaluated by platelet rich plasma (PRP) contact studies and viewed by scanning electron microscopy (SEM) using BioSpan and non-grafted polyurethane as references. We found that fewer platelets adhered to the MPC-grafted surfaces and that they showed less shape variation than the references. These results suggest that these grafted polymers may have the possibility of the usage for biomaterials.

Biocompatible Materials↗

PubMed2002

Regulation by PGE2 of the production of interleukin-6, macrophage colony stimulating factor, and vascular endothelial growth factor in human synovial fibroblasts.

1. We examined the effects of endogenous prostaglandin E(2) (PGE(2)) on the production of interleukin-6 (IL-6), macrophage colony stimulating factor (M-CSF), and vascular endothelial growth factor (VEGF) by interleukin-1beta (IL-1beta)-stimulated human synovial fibroblasts. 2. NS-398 (1 microM), a cyclo-oxygenase-2 (COX-2) inhibitor, inhibited IL-6 and VEGF production (35+/-4% and 26+/-2%, respectively) but enhanced M-CSF production (38+/-4%) by IL-1beta (1 ng ml(-1)) in synovial fibroblasts isolated from patients with osteoarthritis (OA) and rheumatoid arthritis (RA). Exogenous PGE(2) completely abolished the effects of NS-398 on the production of each mediator by OA fibroblasts stimulated with IL-1beta. 3. 8-Bromo cyclic AMP and dibutyryl cyclic AMP, cyclic AMP analogues, mimicked the effects of PGE(2) on IL-6, M-CSF, and VEGF production by OA fibroblasts. 4. The EP(2) selective receptor agonist ONO-AE1-259 (2 nM) and the EP(4) selective receptor agonist ONO-AE1-329 (2 or 20 nM), but not the EP(1) selective receptor agonist ONO-DI-004 (1 microM) and the EP(3) selective receptor agonist ONO-AE-248 (1 microM), replaced the effects of PGE(2) on IL-6, M-CSF, and VEGF production by OA and RA fibroblasts stimulated with IL-1beta in the presence of NS-398. 5. Both OA and RA fibroblasts expressed mRNA encoding EP(2) and EP(4) but not EP(1) receptors. In addition, up-regulation of EP(2) and EP(4) receptor mRNAs was observed at 3 h after IL-1beta treatment. 6. These results suggest that endogenous PGE(2) regulates the production of IL-6, M-CSF, and VEGF by IL-1beta-stimulated human synovial fibroblasts through the activation of EP(2) and EP(4) receptors with increase in cyclic AMP.

Cells, Cultured↗

PubMed2002

RNA cleavage efficiency and catalytic turnover of an oligonucleotide-two copper complexes conjugate.

We synthesized a 2'-O-methyloligonucleotide with two terpyridine-Cu(II) complexes in the central part, and found that this antisense agent cleaves an RNA oligomer more efficiently than an analogous agent with one Cu(II) complex. Determination of the pH dependence of the activity revealed the existence of an optimum pH for the two complexes-agent, which may be a result of the cooperative action of the complexes. In addition, we demonstrated that the novel agent catalytically cleaved an excess of the RNA substrate.

Base Sequence↗

PubMedDate not supplied

Mutant ICR mouse, kuru2, manifests hearing impairment and abnormal behavior.

BACKGROUND: Establishment of mutant animals presents valuable information on corresponding human diseases. We established a mutant mouse, kuru2, from the previously reported Ascites ICR Mouse. MATERIALS AND METHODS: Epileptic individuals of Ascites ICR Mouse were mated and maintained with sibling mating. This mouse was characterized by hearing impairment and abnormal behavior such as ataxic gait, disturbance of positional sense, hyperirritability, head-tossing and circling movement. RESULTS: No detectable auditory brain stem response was evoked from an early stage of life. Abnormal behavior started from 4 to 12 weeks of age. Microscopic examination revealed no major abnormalities in the central nervous system. In the inner ear, the vestibule and cochlea were well developed, however degeneration of the spiral ganglions was observed at a late age. The genetic mode was autosomal recessive. DISCUSSION: Since this mouse has a distinctive phenotype, the animal may provide an understanding of hereditary hearing impairment and abnormal behavior.

Animals↗