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Biomedical subjects

Koichi Ito

Publications and source records attributed to Koichi Ito.

At least 19 recordsLinked to original sources

Loss and Gain of Aqp10 Paralogs With Broad Solute Selectivity in Anguillid Eels.

Aquaporin (Aqp) 10 is a member of the aquaglyceriporin family, which transports small, uncharged solutes in addition to water. Although the solute selectivity of aquaglyceroporins varies, the mechanism of solute selectivity has not yet been fully elucidated. The common ancestor of ray-finned fish possessed two paralogous genes for aquaporin 10, aqp10.1 and aqp10.2, which produce Aqps with different solute selectivities. Most teleosts possess one or more ohnologs derived from aqp10.1 and aqp10.2; however, the common ancestor of Anguilliformes species lost all aqp10.1-derived ohnologs. Anguilliformes species, except Anguilla species, have one aqp10.2b, but recent tandem duplications in the European eel have generated three aqp10.2b paralogs (aqp10.2b1-aqp10.2b3), whose activities remain ambiguous. In this study, we found that the four sites forming the aromatic/arginine (ar/R) selectivity filter in European eel Aqp10.2b1 were identical to those in Aqp10.2b of other species. However, the Y residue at position 3 was replaced with G in the ar/R selectivity filter of Aqp10.2b2 and b3. When expressed in Xenopus oocytes, Aqp10.2b2 and b3 showed higher permeability to urea and boric acid than Aqp10.2b1, indicating that Aqp10.2b2 and b3 acquired broad solute selectivity similar to that of Aqp10.1, which was lost in the ancestral Anguilliformes species. Urea and boric acid permeabilities of Aqp10.2b1 increased when the Y residue at position 3 of the ar/R selectivity filter was replaced with G. Overall, our results outline the history of the loss and gain of Aqp10 paralogs with broad solute selectivity in anguillid eels.

Animals↗

Management of differentiated thyroid carcinoma with radioiodine and recombinant human TSH.

Recombinant human TSH (rhTSH) brought revolutionary change in the management of patients with differentiated thyroid cancer since it was first approved for clinical use in the United States and Europe. Follow-up management of differentiated thyroid cancer is based on the detection of recurrent or residual cancer, traditionally achieved by measurement of serum thyroglobulin level and various imaging techniques including 131I whole body scan. Previously, TSH stimulation was achieved only by induction of hypothyroidism following withdrawal of thyroid hormone. However, hypothyroidism is uncomfortable and is association with a reduction in quality of life. RhTSH can provide elevated TSH without making patients hypothyroid. In the United States and Europe, rhTSH is approved for use only in monitoring of differentiated thyroid cancer. In this article, we reviewed the role of rhTSH in the diagnosis and management of differentiated thyroid cancer.

Carcinoma↗

Effects of low-intensity pulsed ultrasound on the differentiation of C2C12 cells.

Low-intensity pulsed ultrasound (LIPUS) is known to accelerate bone regeneration, but the precise cellular mechanism is still unclear. The purpose of this study was to determine the effect of LIPUS on the differentiation of pluripotent mesenchymal cell line C2C12. The cells were cultured in differentiation medium with or without the addition of LIPUS stimulation. The ultrasound signal consisted of 1.5 MHz at an intensity of 70 mW/cm2 for 20 min for all cultures. To verify the cell lineage after LIPUS stimulation, mRNA expression of cellular phenotype-specific markers characterizing osteoblasts (Runx2, Msx2, Dlx5, AJ18), chondroblasts (Sox9), myoblasts (MyoD), and adipocytes (C/EBP, PPARgamma) was studied using real-time polymerase chain reaction analysis. The protein expression of Runx2 and activated phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinase (p38 MAPK) were performed using Western blotting. The mRNA expression of Runx2, Msx2, Dlx5, AJ18, and Sox9 was increased markedly by the LIPUS stimulation, whereas the expression of MyoD, C/EBP, and PPARgamma was drastically decreased. In the Western blot analysis, LIPUS stimulation increased Runx2 protein expression and phosphorylation of ERK1/2 and p38 MAPK. Our study demonstrated that LIPUS stimulation converts the differentiation pathway of C2C12 cells into the osteoblast and/or chondroblast lineage via activated phosphorylation of ERK1/2 and p38 MAPK.

Animals↗

Prevalence of hepatitis E virus infection in Japanese children.

BACKGROUND: Recently, sporadic cases of acute hepatitis and fulminant hepatitis caused by hepatitis E virus (HEV) have been reported in Japan. However, few reports have addressed the issue of HEV infection during childhood. METHODS: This study included 5 patients with fulminant hepatitis, 30 patients with acute hepatitis, and 309 patients without history of hepatic dysfunction or hepatitis in childhood as control. RNA was extracted from each serum sample, and HEV specific reverse-transcriptase polymerase chain reaction was performed. Anti-HEV immunoglobulin (Ig)M and IgG were measured by enzyme-linked immunoadsorbent assay. RESULTS: HEV RNA, anti-HEV IgM, and anti-HEV IgG were not detected in the sera of any of the five patients with fulminant hepatitis. In the 30 patients with acute hepatitis, only one (3.3%) was positive for anti-HEV IgG, and all were negative for anti-HEV IgM and HEV RNA. Of the 309 control patients, 8 (2.6%) were positive for anti-HEV IgG, and 2 (0.6%) were positive for anti-HEV IgM, respectively. CONCLUSION: The result of patients with fulminant hepatitis suggests that HEV is an unlikely cause of fulminant hepatitis in children. However, the detection rate of anti-HEV IgG shows that a history of HEV infection is not so rare among children in Japan.

Acute Disease↗

Ribosomal protein L11 mutations in two functional domains equally affect release factors 1 and 2 activity.

Bacterial release factors (RFs) 1 and 2 catalyse translation termination at UAG/UAA and UGA/UAA stop codons respectively. It has been shown that limiting the amount of ribosomal protein L11 affects translation termination at UAG and UGA differently. To understand the functional interplay between L11 and RF1/RF2, we isolated 21 distinct mutations in L11 as suppressors of either temperature-sensitive (ts) RF1/RF2 strains or read-through mutants of lacZ nonsense (UAG or UGA) strains. 10 of 21 mutants restored ts lethal growth of RF1 and/or RF2 strains. All the selected L11 mutants, including the RF1ts- and RF2ts-specific suppressors, had the same effect, either enhancing or reducing, on UAG and UGA termination efficiency in vivo. The specific properties of the selected L11 mutations remained unchanged in an RF3 deletion strain. Moreover, ribosomes absent of L11 had equally reduced activity for both RF1- and RF2-mediated peptide release in vitro. These results suggest that, unlike the previous notion, L11 has a common, cooperative role with RF1 and RF2. These L11 mutations were located on the surface of two domains of L11, and interpreted to affect the interaction between L11 and rRNA or the RFs thereby leading to the altered translation termination.

Codon, Terminator↗

Effects of bioactive glass on bone augmentation within a titanium cap in rabbit parietal bone.

BACKGROUND: The condition of alveolar bone influences the success and subsequent esthetics of implant treatment. This study investigated the early effects of bioactive glass (BG) on bone augmentation within a hemispherical titanium cap in rabbit parietal bone. METHODS: Twelve adult male Japanese white rabbits were used. One titanium cap (test site) was packed with BG in a collagen gel, and the other (control site) was packed with the collagen gel alone. After 1 and 3 months, animals were euthanized, and the experimental area was examined using fluorescence and light microscopy. RESULTS: Newly generated bone was observed at 1 and 3 months of healing. Although bone was also generated without BG, newly mineralized bone was generated sooner with BG present for guided bone augmentation than without BG. At 1 and 3 months, the BG was not bioabsorbed completely, and some particles remained. CONCLUSION: New bone is generated at an early stage of bone formation using BG for bone augmentation.

Animals↗

Profiling the cytokines in gingival crevicular fluid using a cytokine antibody array.

BACKGROUND: Various compounds have been detected in gingival crevicular fluid (GCF) as indicators of periodontal disease activity. Therefore, the analysis of GCF may be especially beneficial for diagnosing current periodontal status and addressing the effects of treatment. Moreover, the identification of new markers in GCF may also contribute to elucidating novel mechanisms involved in periodontal disease. This study sought novel marker proteins specific to chronic periodontitis by profiling cytokines in GCF using a cytokine antibody array system. METHODS: Human cytokine array V, which detects 79 cytokines on one membrane, was used to determine the profile of cytokines in GCF from seven subjects with chronic periodontitis and seven subjects with healthy periodontia. The profile was exposed to x-ray film and quantified using image analysis software. Healthy and diseased sites were compared statistically. RESULTS: We detected 10 cytokines in periodontally healthy sites and 36 cytokines in periodontally diseased sites. Interleukin-8 (IL-8) and transforming growth factor-beta 2 (TGF-beta2) were detected at high levels in healthy and diseased subjects. There were significant differences between healthy and diseased subjects in the levels of tissue inhibitor of metalloproteinases-2 (TIMP-2), tumor necrosis factor-beta (TNF-beta), growth-related oncogene (GRO), interferon-inducible protein-10 (IP-10), angiogenin (Ang), vascular endothelial growth factor (VEGF), insulin-like growth factor binding protein-3 (IGFBP-3), osteoprotegerin (OPG), epidermal growth factor (EGF), glial-derived neurotrophic factor (GDNF), pulmonary and activation-regulated chemokine (PARC), oncostatin M (OSM), fibroblast growth factor-4 (FGF-4), IL-16, homologous to lymphotoxins (LIGHT), and placenta growth factor (PlGF). Of these, the newly detected cytokines were GRO, Ang, IGFBP-3, GDNF, PARC, OSM, FGF-4, IL-16, LIGHT, and PlGF. CONCLUSIONS: In this study, we detected several cytokines in GCF using a cytokine antibody array system, including both inflammatory cytokines and various growth factors. Therefore, periodontal disease may participate in the wound healing process and in tissue destruction via the inflammatory process. Our results suggest that the quantification of these cytokines in GCF provides useful information for the diagnosis of periodontal disease status.

Aged↗

Effects of different sizes of beta-tricalcium phosphate particles on bone augmentation within a titanium cap in rabbit calvarium.

This study evaluated the effects of different sizes of beta-TCP particles on bone augmentation within a titanium cap. In 20 rabbits, the calvarium was exposed and a circular groove was prepared. After marrow penetration, a standardized hemispherical titanium cap was placed in the circular grove. The cap was filled with small-sized (100-250 microm) or medium-sized (250-500 microm) beta-TCP particles for the experimental site and without beta-TCP for the control site. After one and three months of healing, the animals were euthanized and examined histologically. There was a statistically significant difference in the amount of mineralized bone generated between the experimental and control groups in the three-month specimens. Furthermore, the medium-sized particles showed significantly more mineralized bone than did the small-sized particles. Based on these findings, we suggested that beta-TCP might be effective for bone formation and that medium-sized particles are more useful than small-sized particles in bone maturation.

Animals↗

Treatment of Miller Class III recessions with enamel matrix derivative (Emdogain) in combination with subepithelial connective tissue grafting.

This report examines the clinical parameters and radiographic characteristics of two patients with Class III gingival recession who were treated with combined connective tissue grafting and root coverage with enamel matrix derivative. Both patients presented with multiple Class III recessions in the mandibular central incisors. Root coverage was achieved by connective tissue grafting in conjunction with the application of enamel matrix derivative. Soft tissue coverage of the root surfaces was achieved clinically, and radiographs showed improvement in the interproximal vertical bone defects.

Adult↗

Childhood thyroid carcinoma with BRAFT1799A mutation shows unique pathological features of poor differentiation.

The prevalence of BRAFT1799A mutation in papillary thyroid carcinomas (PTCs) displays a marked age association: relatively high in adults and exceedingly low in childhood. We report a case of a 12-year-old Japanese female with PTC, the only one case that harbored BRAFT1799A mutation in a series of 46 childhood thyroid cancer tissues. On histology, the findings were so atypical that pathologists had repeatedly examined tumor sections to agree on the diagnosis of poorly differentiated follicular carcinoma. Upon molecular investigation, BRAFT1799A was detected in DNA extracted from paraffin-embedded material, whereas TP53 was wild-type. Since BRAFT1799A is strictly limited to PTCs, immunohistochemical staining for CD15, a specific marker for papillary carcinoma, was performed to verify the diagnosis. A small tumor area with papillary-like structure was stained positive. These findings strongly suggest that this case is a poorly differentiated carcinoma that arose from PTC and implies a possible association of BRAF mutation with dedifferentiated phenotype of PTCs.

Carcinoma, Papillary↗

The effect of IL-1alpha on the expression of matrix metalloproteinases, plasminogen activators, and their inhibitors in osteoblastic ROS 17/2.8 cells.

Interleukin-1 (IL-1) plays key roles in altering bone matrix turnover. This turnover is regulated by matrix metalloproteinases (MMPs), tissue inhibitor of matrix metalloproteinases (TIMPs), and the plasminogen activation system, including tissue-type plasminogen activator (tPA), urokinase-type plasminogen activator (uPA) , and plasminogen activator inhibitor type-1 (PAI-1). In this study, we examined the effect of IL-1alpha on the expression of the MMPs, TIMPs, tPA, uPA, and PAI-1 genes in osteoblasts derived from the rat osteosarcoma cell line ROS 17/2.8. The cells were cultured in alpha-minimum essential medium containing 10% fetal bovine serum with 0 or 100 U/ml of IL-1alpha for up to 14 days. The levels of MMPs, TIMPs, uPA, tPA, and PAI-1 expression were estimated by determining the mRNA levels using real-time RT-PCR and by determining protein levels using ELISA. In IL-1alpha cultures, the expression levels of MMP-1, -2, -3, -13, and -14 exceeded that of the control through day 14 of culture, and the expression of MMPs increased markedly from the proliferative to the later stages of culture. The TIMP-1, -2, and -3 expression levels increased from the initial to the proliferative stages of culture. The expression of tPA increased greatly during the proliferative stage of culture, and uPA expression increased throughout the culture period, increasing markedly from the proliferative to the later stages of culture. In contrast, PAI-1 expression decreased in the presence of IL-1alpha through day 14. These results suggest that IL-1alpha stimulate bone matrix turnover by increasing MMPs, tPA, and uPA production and decreasing PAI-1 production by osteoblasts, and incline the turnover to the resolution.

Animals↗

Novel hydrogel membrane based on copoly(hydroxyethyl methacrylate/p-vinylbenzyl-poly(ethylene oxide)) for biomedical applications: properties and drug release characteristics.

The aim of this study was to synthesize and characterize a novel biocompatible polymeric membrane system and demonstrate its potential use in various biomedical applications. Synthetic hydrogels based on poly(hydroxyethyl methacrylate), poly(HEMA), have been widely studied and used in biomedical fields. A novel copolymer hydrogel was prepared in the membrane form using 2-hydroxyethyl methacrylate monomer (HEMA) and a macromonomer p-vinylbenzyl-poly(ethylene oxide) (V-PEO) via photoinitiated polymerization. A series of poly(HEMA/V-PEO) copolymer membranes with different compositions was prepared. The membranes were characterized using infrared, thermal and SEM analysis. The thermal stabilities of the copolymer membranes were found to be lowered by an increase in the ratio of macromonomer (V-PEO) in the membrane structure. Because of the incorporation of PEO segments, the copolymers exhibited significantly higher hydrophilic surface properties than pure poly(HEMA), as demonstrated by contact angle measurements. Equilibrium swelling studies were conducted to investigate the swelling behavior of the membranes. The equilibrium water uptake was reached in about 4 h. Moreover, the blood protein adsorption and platelet adhesion were significantly reduced on the surface of the PEO containing copolymer membranes compared to control pure poly(HEMA). Drug release experiments were performed in a continuous release system using model drug (vancomycin) loaded copoly(HEMA/V-PEO) membranes. A specific poly(HEMA/V-PEO) membrane formulation possessing the highest PEO content (with a HEMA:V-PEO (mmol:mmol) feed ratio of 112:1 and loaded with 40 mg antibiotic/g polymer) released about 81% of the total loaded drug in 24 h at pH 7.4. This membrane composition provided the best results and can be considered as a potential candidate for a transdermal antibiotic carrier and various biomedical and biotechnological applications.

Anti-Bacterial Agents↗

Divergent pharmacological activity of novel marine-derived excitatory amino acids on glutamate receptors.

Kainate receptors show a particular affinity for a variety of natural source compounds, including dysiherbaine (DH), a potent agonist derived from the marine sponge Dysidea herbacea. In this study, we characterized the pharmacological activity and structural basis for subunit selectivity of neodysiherbaine (neoDH) and MSVIII-19, which are natural and synthetic analogs of DH, respectively. NeoDH and MSVIII-19 differ from DH in the composition of two functional groups that confer specificity and selectivity for ionotropic glutamate receptors. In radioligand binding assays, neoDH displayed a 15- to 25-fold lower affinity relative to that of DH for glutamate receptor (GluR)5 and GluR6 kainate receptor subunits but a 7-fold higher affinity for kainate (KA)2 subunits, whereas MSVIII-19 displaced [(3)H]kainate only from GluR5 subunits but not GluR6 or KA2 subunits. NeoDH was an agonist for kainate and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in patch-clamp recordings; in contrast, MSVIII-19 acted as a potent antagonist for homomeric GluR5 receptor currents with weaker activity on other kainate and AMPA receptors. Neither neoDH nor MSVIII-19 activated group I metabotropic GluRs. Homology modeling suggests that two critical amino acids confer the high degree of selectivity between the dysiherbaine analogs and the GluR5 and KA2 subunits. In summary, these data describe the pharmacological activity of two new compounds, one of which is a selective GluR5 receptor antagonist that will be of use for understanding native receptor function and designing more selective ligands for kainate receptors.

Alanine↗

IL-1 alpha stimulates the formation of osteoclast-like cells by increasing M-CSF and PGE2 production and decreasing OPG production by osteoblasts.

Interleukin-1alpha (IL-1alpha) is one of the most potent bone-resorbing factors involved in the bone loss that is associated with inflammation. We examined the effect of the inflammatory mediator IL-1alpha on the expression of macrophage colony-stimulating factor (M-CSF), osteoprotegerin (OPG), and prostaglandin E2 (PGE2) in rat osteoblasts, and the indirect effect of IL-1alpha on the formation of osteoclast-like cells. Osteoblasts were cultured in alpha-minimum essential medium containing 10% fetal bovine serum with or without 100 units/ml of IL-1alpha for up to 14 days. The gene and protein expression of M-CSF and OPG were estimated by determining mRNA levels using the real-time polymerase chain reaction and protein levels using Western blot analysis. PGE2 expression was determined using an enzyme-linked immunosorbent assay. The formation of osteoclast-like cells was estimated using tartrate-resistant acid phosphatase (TRAP) staining of osteoclast precursors in culture with conditioned medium from IL-1alpha-treated osteoblasts and the soluble receptor activator of NF-kappaB ligand (RANKL). M-CSF and PGE2 expression in osteoblasts increased markedly in cells cultured with IL-1alpha, whereas OPG expression decreased. The conditioned medium containing M-CSF and PGE2 produced by IL-1alpha-treated osteoblasts and soluble RANKL increased the TRAP staining of osteoclast precursors. These results suggest that IL-1alpha stimulated the formation of osteoclast-like cells via an increase in M-CSF and PGE2 production, and a decrease in OPG production by osteoblasts.

Animals↗

Enamel matrix derivative is a potent inhibitor of breast cancer cell attachment to bone.

This study examined whether enamel matrix derivative (EMD) inhibits the adhesion of cancer cells to bone. A typical breast cancer cell line, MCF-7, was used. Conditioned human osteosarcoma cell (Saos-2) medium was used as extracellular bone matrix (ECBM) to measure cell attachment. MCF-7 cells were incubated on ECBM-coated culture plates with or without soluble EMD, Arg-Gly-Asp (RGD) sequence blocking peptides, recombinant bone sialoprotein (rBSP), or specific integrin antibodies, and the attached cells were quantified using toluidine blue staining. EMD markedly reduced the attachment of MCF-7 cells to ECBM in a dose-dependent manner. An RGD peptide (GRGDSP) and recombinant BSP inhibited cell attachment to the same degree as EMD. Similarly, anti-alphavbeta3 integrin antibody strongly reduced cell attachment, whereas anti-alphavbeta5 and anti-beta1 integrin antibodies had less marked effects on cell attachment. These results show that EMD inhibits MCF-7 cell attachment to a bone matrix and that it might be useful as an anti-adhesive agent for breast cancer cells to bone in vivo.

Antibodies, Monoclonal↗