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Biomedical subjects

Makoto Inoue

Publications and source records attributed to Makoto Inoue.

At least 37 records · Page 2Linked to original sources
PubMed2006

Rehmannia glutinosa induces glial cell line-derived neurotrophic factor gene expression in astroglial cells via cPKC and ERK1/2 pathways independently.

Among four herbs of traditional Chinese medicines (TCMs) used in the therapy of dementia, Rehmannia glutinosa (RG) was found to induce the gene expression of glial cell line-derived neurotrophic factor (GDNF) in C6 glioblastoma cells and primary cultured astrocytes. The RG-induced GDNF mRNA up-regulation in C6 glioblastoma cells was completely attenuated by the presence of a pan-specific protein kinase C (PKC) inhibitor (Ro-31-8220) and a MAPK/ERK kinase 1 (MEK1) inhibitor (U0126). A conventional PKC inhibitor (Gö6976) also significantly decreased GDNF gene induction. On the other hand, RG treatment was found to stimulate phosphorylation of extracellular signal-regulated kinase 1 and 2 (ERK1/2), which preceded GDNF mRNA induction in C6 glioblastoma cells. However, none of the PKC inhibitors significantly changed RG-stimulated ERK1/2 phosphorylation. Therefore, RG-stimulated GDNF gene expression could be independently up-regulated through cPKC and ERK 1/2 pathways in C6 glioblastoma cells.

Alzheimer Disease↗
PubMed2006

An Arabidopsis SBP-domain fragment with a disrupted C-terminal zinc-binding site retains its tertiary structure.

SQUAMOSA promoter-binding proteins (SBPs) form a major family of plant-specific transcription factors, mainly related to flower development. SBPs share a highly conserved DNA-binding domain of approximately 80 amino acids (SBP domain), which contains two non-interleaved zinc-binding sites formed by eight conserved Cys or His residues. In the present study, an Arabidopsis SPL12 SBP-domain fragment that lacks a Cys residue involved in the C-terminal zinc-binding pocket was found to retain a folded structure, even though only a single Zn2+ ion binds to the fragment. Solution structure of this fragment determined by NMR is very similar to the previously determined structures of the full SBP domains of Arabidopsis SPL4 and SPL7. Considering the previous observations that chelating all the Zn2+ ions of SBPs resulted in the complete unfolding of the structure and that a mutation of the Cys residue equivalent to that described above impaired the DNA-binding activity, we propose that the Zn2+ ion at the N-terminal site is necessary to maintain the overall tertiary structure, while the Zn2+ ion at the C-terminal site is necessary for the DNA binding, mainly by guiding the basic C-terminal loop to correctly fit into the DNA groove.

Amino Acid Sequence↗
PubMed2006

NSAID zaltoprofen possesses novel anti-nociceptive mechanism through blockage of B2-type bradykinin receptor in nerve endings.

Zaltoprofen, a propionic acid derivative of non-steroidal anti-inflammatory drugs (NSAIDs), was shown to have more powerful inhibitory effects to bradykinin (BK)-nociception than other NSAIDs. However, the molecular mechanisms underlying this potent analgesia are not yet fully understood. Here we attempted to clarify the molecular mechanism underlying zaltoprofen-induced analgesia on BK-induced nociception by a novel algogenic-induced paw flexion (APF) test in mice. The intraplantar (i.pl.) injection of zaltoprofen at 1nmol showed strong analgesic action on BK (i.pl.)-induced nociceptive flexor responses, whereas loxoprofen or its active metabolite loxoprofen-SRS did not. Zaltoprofen also inhibited the nociception induced by [Tyr8]-BK, a specific agonist of B2-type BK receptor, but did not affect the nociception by [Lys-des-Arg9]-BK, a specific agonist of B1-type BK receptor. However, zaltoprofen did not affect the substance P-induced nociception, which is mediated by common post-receptor signaling through nociceptive fibers with BK-ones. All these results suggest that NSAID zaltoprofen possesses novel anti-nociceptive mechanism, which inhibits B2-type BK receptor function in nerve endings.

Analgesics↗
PubMed2006

Naked Sendai virus vector lacking all of the envelope-related genes: reduced cytopathogenicity and immunogenicity.

BACKGROUND: Sendai virus (SeV) is a new class of cytoplasmic RNA vector that is free from genotoxicity that infects and multiplies in most mammalian cells, and directs high-level transgene expression. We improved the vector by deleting all of the envelope-related genes from the SeV genome and thus reducing its immunogenicity. METHODS: The matrix (M), fusion (F) and hemagglutinin-neuraminidase (HN) genes-deleted SeV vector (SeV/DeltaMDeltaFDeltaHN) was recovered in a newly established packaging cell line. Then, the generated SeV/DeltaMDeltaFDeltaHN vector was characterised by comparing with single gene-deleted type SeV vectors. RESULTS: This SeV/DeltaMDeltaFDeltaHN vector carrying the green fluorescent protein gene in place of the envelope-related genes could be propagated to a titer of more than 10(8) cell infectious units/ml. This vector showed an efficient transduction capability in vitro and in vivo, and the cytopathic effect and induction of neutralizing antibody in vivo were greatly reduced compared with those of single gene-deleted type SeV vectors. No activity of neutralizing antibody or anti-HN antibody was seen when SeV/DeltaMDeltaFDeltaHN was transduced ex vivo. Additional introduction of amino acid mutations that had been identified from SeV strains causing persistent infections was also effective for the reduction of cytopathic effects. CONCLUSIONS: The deletion of genes from the SeV genome and the additional mutation are very effective for reducing both the immunogenic and cytopathic reactions to the SeV vector. These modifications are expected to improve the safety and broaden the range of clinical applications of this new class of cytoplasmic RNA vector.

Animals↗
PubMed2006

Subjective visual sensation during vitrectomy under retrobulbar anesthesia.

PURPOSE: To report a precise illustrated image of patient's view during vitrectomy. DESIGN: Interventional case report. METHODS: A 56-year-old male with cystoid macular edema (CME) associated with epiretinal membrane underwent vitrectomy. The patient seemed to find gratification in being operated on because he was able to see what was going on. He was a professional artist in computer graphics, and he drew three different pictures depending on the surgical procedures. RESULTS: These pictures depicted the moving of pasty, whirling fluid during core vitrectomy, numerous black or gray spots like snowflakes by intravitrealy injected crystals of triamcinolone acetonide, and a sharp tapered instrument coming into the center and membrane-like material being peeled off. CONCLUSIONS: Patients may see a precise shape in a constant size, scale, and detailed movement as well, focused on the retina without described optics. Further investigation will be required to determine this mechanism.

Anesthesia, Local↗
PubMed2006

Solution structure of the SWIRM domain of human histone demethylase LSD1.

SWIRM is an evolutionarily conserved domain involved in several chromatin-modifying complexes. Recently, the LSD1 protein, which bears a SWIRM domain, was found to be a demethylase for Lys4-methylated histone H3. Here, we report a solution structure of the SWIRM domain of human LSD1. It forms a compact fold composed of 6 alpha helices, in which a 20 amino acid long helix (alpha4) is surrounded by 5 other short helices. The SWIRM domain structure could be divided into the N-terminal part (alpha1-alpha3) and the C-terminal part (alpha4-alpha6), which are connected to each other by a salt bridge. While the N-terminal part forms a SWIRM-specific structure, the C-terminal part adopts a helix-turn-helix (HTH)-related fold. We discuss a model in which the SWIRM domain acts as an anchor site for a histone tail.

Amino Acid Motifs↗
PubMed2006

Solution structures of the SURP domains and the subunit-assembly mechanism within the splicing factor SF3a complex in 17S U2 snRNP.

The SF3a complex, consisting of SF3a60, SF3a66, and SF3a120, in 17S U2 snRNP is crucial to spliceosomal assembly. SF3a120 contains two tandem SURP domains (SURP1 and SURP2), and SURP2 is responsible for binding to SF3a60. We found that the SURP2 fragment forms a stable complex with an SF3a60 fragment (residues 71-107) and solved its structure by NMR spectroscopy. SURP2 exhibits a fold of the alpha1-alpha2-3(10)-alpha3 topology, and the SF3a60 fragment forms an amphipathic alpha helix intimately contacting alpha1 of SURP2. We also solved the SURP1 structure, which has the same fold as SURP2. The protein-binding interface of SURP2 is quite similar to the corresponding surface of SURP1, except for two amino acid residues. One of them, Leu169, is characteristic of SF3a120 SURP2 among SURP domains. Mutagenesis showed that this single Leu residue is the critical determinant for complex formation, which reveals the protein recognition mechanism in the subunit assembly.

Amino Acid Sequence↗
PubMed2006

Solution structure of the kinase-associated domain 1 of mouse microtubule-associated protein/microtubule affinity-regulating kinase 3.

Microtubule-associated protein/microtubule affinity-regulating kinases (MARKs)/PAR-1 are common regulators of cell polarity that are conserved from nematode to human. All of these kinases have a highly conserved C-terminal domain, which is termed the kinase-associated domain 1 (KA1), although its function is unknown. In this study, we determined the solution structure of the KA1 domain of mouse MARK3 by NMR spectroscopy. We found that approximately 50 additional residues preceding the previously defined KA1 domain are required for its proper folding. The newly defined KA1 domain adopts a compact alpha+beta structure with a betaalphabetabetabetabetaalpha topology. We also found a characteristic hydrophobic, concave surface surrounded by positively charged residues. This concave surface includes the highly conserved Glu-Leu-Lys-Leu motif at the C terminus, indicating that it is important for the function of the KA1 domain.

Amino Acid Sequence↗
PubMed2006

Immunohistochemical detection of myogenin and p21 in methylcholanthrene-induced mouse rhabdomyosarcomas.

3-Methylcholanthrene (MC)-induced 10 embryonal (ERSs) and 24 pleomorphic rhabdomyosarcomas (PRSs) of the dermis in mouse were examined immunohistochemically for myogenin, p21 and proliferating cell nuclear antigen (PCNA) nuclear reactivity and myosin reactivity. ERSs had higher expression of myogenin and p21 compared with that of myosin. PRSs were divided into two groups having high (moderate or marked reactivity; HLM) and low (mild reactivity; LLM) levels of myosin expression. Expression of p21 was higher in HLM-PRSs than in LLM-PRSs. Statistically significant association was observed between myosin and p21 expression in PRSs, but not between myosin and myogenin expression. Myogenin and p21 reactivity were observed in myoblast-like cells, but rarely in multinucleated cells. In ERSs, small undifferentiated myogenic precursor cells were also positive for p21. No difference of PCNA reactivity was observed between HLM-PRSs and LLM-PRSs, although its reactivity was higher in PRSs than in ERSs. The results suggest that myogenin is related to myoblast-like cell differentiation in PRSs and that p21 plays essential roles in myotube formation and myosin expression. In ERSs, p21 may be involved in inhibition of myogenic precursor cell proliferation and differentiation.

Animals↗
PubMed2006

Psychological impact of verbal abuse and violence by patients on nurses working in psychiatric departments.

The aim of this study was to assess the psychological impact of verbal abuse or violence by patients on nurses working in psychiatry departments and to identify factors related to their impact. Survey sheets were distributed to a total of 266 nurses working at two hospitals, and replies were obtained from 232 of them. Because three of them had less than 1 month of experience working in the psychiatry department and four of them failed to answer all the questions, valid replies were obtained from 225 nurses. Among the 225 whose replies were valid, 141 nurses who replied that they had experienced verbal abuse or violence that left an impression on them remained as the subjects of the final analysis. The Impact of Event Scale-Revised (IES-R) was used to evaluate psychological impact. Of the nurses who had been exposed to verbal abuse or violence that left an impression, 21% had scores that exceeded the IES-R cut-off point (24/25), and low satisfaction with family support, and neuroticism on the Eysenck Personality Questionnaire-Revised were shown to have contributed to the psychological impact. Nurses working in psychiatry departments were shown to experience a severe psychological impact when exposed to verbal abuse or violence. These results suggest the need for mental care approaches for nurses working in psychiatry departments.

Adaptation, Psychological↗
PubMed2006

Suppression of ocular inflammation in endotoxin-induced uveitis by inhibiting nonproteolytic activation of prorenin.

PURPOSE: A recent study revealed that angiotensin receptor signaling mediates ocular inflammation and neovascularization. It was also found that prorenin undergoes nonproteolytic activation leading to upregulation of the renin-angiotensin system (RAS) when prorenin receptor interacts specifically with the handle region of prorenin. The purpose of the present study was to elucidate the role of the receptor-dependent nonproteolytic activation of prorenin in ocular inflammation in endotoxin-induced uveitis (EIU). METHODS: EIU was induced in Long-Evans rats by a single intraperitoneal injection of 100 microg lipopolysaccharide (LPS). Tissue localization of total prorenin, prorenin receptor, and activated prorenin in the EIU retina was examined by immunohistochemistry. To inhibit the prorenin receptor-mediated upregulation of the RAS, a decoy handle-region peptide (HRP) was intraperitoneally administered 24 hours before and immediately after the injection of LPS. Twenty-four hours after LPS injection, leukocyte adhesion to the retinal vasculature was evaluated with a concanavalin A lectin perfusion-labeling technique. In addition, leukocyte infiltration into the vitreous cavity and protein concentration in the anterior chamber were also measured. Retinal mRNA and protein levels of intercellular adhesion molecule (ICAM)-1, interleukin (IL)-6, and C-C chemokine ligand (CCL) 2/monocyte chemotactic protein (MCP)-1 were examined by RT-PCR and ELISA. RESULTS: Retinal vessels in rats with EIU were strongly positive for total prorenin, prorenin receptor, and activated prorenin. Systemic treatment with HRP resulted in dose- and time-dependent inhibition of the leukocyte adhesion and infiltration and the protein leakage, all of which were increased by the induction of EIU. Retinal mRNA expression and protein levels of ICAM-1, CCL2/MCP-1 and IL-6, induced in rats with EIU, were also significantly suppressed with application of HRP. CONCLUSIONS: These findings demonstrate for the first time that nonproteolytically activated prorenin plays a significant role in the development of ocular inflammation in the EIU model. The present study suggests the potential use of HRP, a decoy peptide binding to the prorenin receptor, as a therapeutic agent to reduce ocular inflammation.

Animals↗
PubMed2006

Neuroprotective effects of angiotensin II type 1 receptor (AT1R) blocker, telmisartan, via modulating AT1R and AT2R signaling in retinal inflammation.

PURPOSE: To investigate the retinal neural damage that occurs during inflammation and the therapeutic effects of the angiotensin II type 1 receptor (AT1R) blocker, telmisartan, using a model of endotoxin-induced uveitis (EIU). METHODS: The localization of AT1R and AT2R was shown by immunohistochemistry. EIU was induced by intraperitoneal injection of lipopolysaccharide (LPS). Animals were treated with telmisartan for 2 days and were evaluated 24 hours later. Expression levels of angiotensin II, STAT3 activation induced by inflammatory cytokines, and retinal proteins essential for neural activities (e.g., synaptophysin, rhodopsin) were analyzed by immunoblot. An AT2R antagonist was administered to evaluate the contribution of AT2R signaling in this therapy. Dark-adapted full-field electroretinography (ERG) was also performed. RESULTS: AT1R and AT2R were expressed in presynaptic terminals in most of the retinal neurons. AT1R was also expressed in Müller glial cells. During inflammation, angiotensin II expression was elevated, STAT3 was activated, and synaptophysin and rhodopsin expression were reduced. The expression of glial fibrillary acidic protein (GFAP), downstream of STAT3 activation, was induced in Müller glial cells. However, treatment with telmisartan successfully avoided all these changes. An AT2R antagonist lowered synaptophysin expression despite the treatment. STAT3 activity was negatively correlated with rhodopsin expression. Furthermore, ERG responses, which were mostly prevented by telmisartan, were disturbed during inflammation. CONCLUSIONS: Retinal protein expression and visual function are both disturbed by inflammation. Treatment with the AT1R blocker telmisartan efficiently prevented these signs of retinal neural damage through the reduction of local angiotensin II expression, the blockade of AT1R, and the relative upregulation of AT2R function.

Angiotensin II Type 1 Receptor Blockers↗
PubMed2006

Islet-like cell clusters: viability, cell types, and subretinal transplantation in pancreatectomized cats.

We investigated a method for isolating sufficient feline islets of Langerhans to restore normoglycaemia following transplantation into the subretinal space of pancreatectomized cats. Collagenase digestate of feline pancreas was maintained in serum-free tissue culture medium for 1-9 days. Viability of islet-like cell clusters (ICC) was assessed with ethidium bromide and fluorescein diacetate staining; cell types were identified immunohistochemically. After nine days, the ICC were transplanted. We estimated viable ICC in tissue culture at nine days as 3800 +/- 2000 (mean +/- SD) per pancreas. While numbers of beta cells decreased over time in culture, ductal cells increased. Bromodeoxyuridine labelling showed no proliferation of beta cells but extensive proliferation of ductal cells. Subretinal transplants of cultured ICC in the diabetic cats maintained normoglycaemia for up to 12 days, while they provoked massive lymphocyte infiltration indicating rejection. Islet transplantation into the feline subretinal space temporarily restored normoglycaemia. Our current method of culture achieved sufficient reduction of acinar cells but an insufficient yield of insulin-producing cells.

Animals↗
PubMed2006

Immunohistochemical detection of Mdm2 and p53 in feline mammary gland tumors.

The objective of this study was to evaluate nuclear reactivity of Mdm2 and p53 proteins by immunohistochemical means in feline mammary gland tumors; 12 adenomas which included 6 adenomatous lesions obtained from the tissue adjacent to adenocarcinomas, and 22 adenocarcinomas. Seven adenomas and 18 adenocarcinomas showed moderate or marked Mdm2 reactivity. Sixteen adenocarcinomas showed moderate to marked p53 reactivity, but 9 adenomas showed none. Discordant Mdm2 overexpression was found in 5 adenomas and 3 adenocarcinomas, although co-overexpression of Mdm2 and p53 was found in 15 adenocarcinomas. These results suggest that nuclear overexpression of Mdm2 is present in the tumors of early stage without p53 overexpression and related to feline mammary gland tumorigenesis. Nuclear overexpression of p53 is more frequent in adenocarcinomas, but not in adenomas.

Adenocarcinoma↗
PubMed2006

Immunohistochemical analysis for Mdm2 and p53 proteins in methylcholanthrene-induced mouse rhabdomyosarcomas.

3-methylcholanthrene (MC)-induced mouse 10 embryonal (ERSs) and 24 pleomorphic rhabdomyosarcomas (PRSs) of the dermis were examined immunohistochemically for nuclear reactivity of Mdm2, p53, and proliferating cell nuclear antigen (PCNA). ERSs were microscopically present in the rhabdium layer of the dermis from 10 to 13 weeks post injection (PI), and PRSs developed from 13 weeks PI. Moderate to marked Mdm2 reactivity was observed in each of the 10 ERSs, and 23 of the 24 PRSs. Moderate to marked p53 reactivity was observed in 5 of the 10 ERSs, and 19 of the 24 PRSs. p53 reactivity increased in PRSs compared with ERSs. The level of Mdm2 expression was significantly higher compared with p53 expression. Discordant Mdm2 overexpression was observed in 5 ERSs and 5 PRSs, and discordant p53 overexpression was observed in 1 PRSs, although co-overexpression of Mdm2 and p53 was observed in 5 ERSs and 18 PRSs. PCNA reactivity significantly increased in PRSs compared with ERSs. These results suggest that Mdm2 overexpression is an important pathogenic event in MC-induced mouse rhabdomyosarcomas, and its expression may be induced by p53-independent pathway.

Animals↗
PubMed2006

Immunohistochemical detection of p27 and p21 proteins in canine hair follicle and epidermal neoplasms.

Trichoblastomas, trichoepitheliomas, and squamous cell carcinomas in the skin of dogs were analysed by immunohistochemistry for the nuclear expression of p27, p21 and proliferating cell nuclear antigen (PCNA). High levels of p27 were present in trichoepitheliomas and trichoblastomas compared with squamous cell carcinomas. Detectable p21 was found in trichoepitheliomas and squamous cell carcinomas, but trichoblastomas had low level of p21 nuclear reactivity. Low levels of PCNA were detected in trichoepitheliomas and trichoblastomas compared with squamous cell carcinomas. The results suggested that nuclear p27 acts as a cyclin-dependent kinase (CDK) inhibitor in trichoepitheliomas and trichoblastomas. Nuclear p21 expression is involved in the induction of epithelial differentiation and seems to be unrelated to CDK inhibition.

Animals↗