37 results on '"Wada, Y."'
Search Results
2. Novel System for Monitoring Autophagy in the Yeast Saccharomyces cerevisiae
- Author
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Noda, T., primary, Matsuura, A., additional, Wada, Y., additional, and Ohsumi, Y., additional
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- 1995
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3. The Molecular Mechanism of the Induction of the Low Density Lipoprotein Receptor by Chenodeoxycholic Acid in Cultured Human Cells
- Author
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Kawabe, Y., primary, Shimokawa, T., additional, Matsumoto, A., additional, Honda, M., additional, Wada, Y., additional, Yazaki, Y., additional, Endo, A., additional, Itakura, H., additional, and Kodama, T., additional
- Published
- 1995
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4. Lupus Anticoagulant Autoantibody Induces Apoptosis in Umbilical Vein Endothelial Cells: Involvement of Annexin V
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Nakamura, N., primary, Shidara, Y., additional, Kawaguchi, N., additional, Azuma, C., additional, Mitsuda, N., additional, Onishi, S., additional, Yamaji, K., additional, and Wada, Y., additional
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- 1994
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5. Sterol-Mediated Regulation of SREBP-1a,1b,1c and SREBP-2 in Cultured Human Cells
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Kawabe, Y., primary, Honda, M., additional, Wada, Y., additional, Yazaki, Y., additional, Suzuki, T., additional, Ohba, Y., additional, Nabata, H., additional, Endo, A., additional, Matsumoto, A., additional, Itakura, H., additional, and Kodama, T., additional
- Published
- 1994
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6. Therapeutic Effects of Imidazolineoxyl N-Oxide Against Endotoxin Shock through Its Direct Nitric Oxide-Scavenging Activity
- Author
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Yoshida, M., primary, Akaike, T., additional, Wada, Y., additional, Sato, K., additional, Ikeda, K., additional, Ueda, S., additional, and Maeda, H., additional
- Published
- 1994
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7. Erratum
- Author
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Saito, H., primary, Nishikawa, A., additional, Gu, J.G., additional, Ihara, Y., additional, Soejima, H., additional, Wada, Y., additional, Sekiya, C., additional, Niikawa, N., additional, and Taniguchi, N., additional
- Published
- 1994
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8. CDNA Cloning and Chromosomal Mapping of Human N-Acetylglucosaminyltransferase-V
- Author
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Saito, H., primary, Nishikawa, A., additional, Gu, J.G., additional, Ihara, Y., additional, Soejima, H., additional, Wada, Y., additional, Sekiya, C., additional, Niikawa, N., additional, and Taniguchi, N., additional
- Published
- 1994
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9. Lysine demethylase 2B regulates angiogenesis via Jumonji C dependent suppression of angiogenic transcription factors.
- Author
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Sasaki Y, Higashijima Y, Suehiro JI, Sugasawa T, Oguri-Nakamura E, Fukuhara S, Nagai N, Hirakawa Y, Wada Y, Nangaku M, and Kanki Y
- Subjects
- Cell Proliferation, Endothelial Cells metabolism, Humans, Jumonji Domain-Containing Histone Demethylases genetics, Jumonji Domain-Containing Histone Demethylases metabolism, Lysine metabolism, Transcription Factors metabolism, Vascular Endothelial Growth Factor A genetics, Vascular Endothelial Growth Factor A metabolism, F-Box Proteins genetics, F-Box Proteins metabolism, Histones metabolism
- Abstract
Vascular endothelial growth factor (VEGF) signaling plays a central role in vascular development and maintenance of vascular homeostasis. In endothelial cells (ECs), VEGF activates the gene expression of angiogenic transcription factors (TFs), followed by induction of downstream angiogenic responsive genes. Recent findings support that histone modification dynamics contribute to the transcriptional control of genes that are important for EC functions. Lysine demethylase 2B (KDM2B) demethylates histone H3K4me3 and H3K36me2/3 and mediates the monoubiquitination of histone H2AK119. KDM2B functions as a transcriptional repressor in somatic cell reprogramming and tumor development. However, the role of KDM2B in VEGF signaling remains to be elucidated. Here, we show that KDM2B knockdown enhances VEGF-induced angiogenesis in cultured human ECs via increased migration and proliferation. In contrast, ectopic expression of KDM2B inhibits angiogenesis. The function of KDM2B may depend on its catalytic Jumonji C domain. Genome-wide analysis further reveals that KDM2B selectively controls the transcription of VEGF-induced angiogenic TFs that are associated with increased H3K4me3/H3K36me3 and decreased H2AK119ub. These findings suggest an essential role of KDM2B in VEGF signaling in ECs. As dysregulation of VEGF signaling in ECs is involved in various diseases, including cancer, KDM2B may be a potential therapeutic target in VEGF-mediated vasculopathic diseases., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2022
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10. First-in-human assessment of the novel LAT1 targeting PET probe 18 F-FIMP.
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Nozaki S, Nakatani Y, Mawatari A, Hume WE, Wada Y, Ishii A, Tanaka M, Tsuyuguchi N, Doi H, and Watanabe Y
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- Adult, Brain Neoplasms diagnostic imaging, Brain Neoplasms pathology, Female, Fluorodeoxyglucose F18 pharmacokinetics, Glioblastoma diagnostic imaging, Glioblastoma metabolism, Glioblastoma pathology, Glioma diagnostic imaging, Glioma metabolism, Glioma pathology, Humans, Male, Molecular Probes pharmacokinetics, Radiopharmaceuticals metabolism, Radiopharmaceuticals pharmacokinetics, Reproducibility of Results, Sensitivity and Specificity, Tissue Distribution, Brain Neoplasms metabolism, Fluorodeoxyglucose F18 metabolism, Large Neutral Amino Acid-Transporter 1 metabolism, Molecular Probes metabolism, Positron-Emission Tomography methods
- Abstract
In the first-in-human PET study, we evaluated the biodistribution and tumor accumulation of the novel PET probe, (S)-2-amino-3-[3-(2-
18 F-fluoroethoxy)-4-iodophenyl]-2-methylpropanoic acid (18 F-FIMP), which targets the tumor-related L-type amino acid transporter 1 (LAT1), and compared it with L-[methyl-11 C]methionine (11 C-MET) and 2-Deoxy-2-18 F-fluoro-D-glucose (18 F-FDG).18 F-FIMP biodistribution was revealed by whole-body and brain scans in 13 healthy controls. Tumor accumulation of18 F-FIMP was evaluated in 7 patients with a brain tumor, and compared with those of11 C-MET and18 F-FDG. None of the subjects had significant problems due to probe administration, such as adverse effects or abnormal vital signs.18 F-FIMP was rapidly excreted from the kidneys to the urinary bladder. There was no characteristic physiological accumulation in healthy controls.18 F-FIMP PET resulted in extremely clear images in patients with suspected glioblastoma compared with11 C-MET and18 F-FDG.18 F-FIMP could be a useful novel PET probe for LAT1-positive tumor imaging including glioblastoma., Competing Interests: Declaration of competing interest All authors declare no competing interests., (Copyright © 2022 Elsevier Inc. All rights reserved.)- Published
- 2022
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11. Activation of α7 nicotinic acetylcholine receptors attenuates monocyte-endothelial adhesion through FUT7 inhibition.
- Author
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Wu CH, Inoue T, Nakamura Y, Uni R, Hasegawa S, Maekawa H, Sugahara M, Wada Y, Tanaka T, Nangaku M, and Inagi R
- Subjects
- Benzylidene Compounds pharmacology, Cell Adhesion drug effects, Cell Adhesion genetics, Down-Regulation drug effects, Down-Regulation genetics, Fucosyltransferases metabolism, Gene Knockdown Techniques, Human Umbilical Vein Endothelial Cells drug effects, Human Umbilical Vein Endothelial Cells metabolism, Humans, Monocytes drug effects, Monocytes metabolism, Pyridines pharmacology, U937 Cells, alpha7 Nicotinic Acetylcholine Receptor antagonists & inhibitors, Fucosyltransferases antagonists & inhibitors, Human Umbilical Vein Endothelial Cells cytology, Monocytes cytology, alpha7 Nicotinic Acetylcholine Receptor metabolism
- Abstract
Cholinergic anti-inflammatory pathway (CAP) describes a neuronal-inflammatory reflex centered on systemic cytokine regulation by α7 nicotinic acetylcholine receptor (α7nAChR) activation of spleen-residue macrophage. However, the CAP mechanism attenuating distal tissue inflammation, inducing a low level of systemic inflammation, is lesser known. In this study, we hypothesized that CAP regulates monocyte accessibility by influencing their adhesion to endothelial cells. Using RNA-seq analysis, we identified that α1,3-Fucosyltransferase 7 (FucT-VII), the enzyme required for processing selectin ligands, was significantly downregulated by α7nAChR agonist among other cell-cell adhesion genes. The α7nAChR agonist inhibited monocytic cell line U-937 binding to P-selectin and adhesion to endothelial cells. Furthermore, α7nAChR agonist selectivity was confirmed by α7nAChR knockdown assays, showing that FUT7 inhibition and adhesion attenuation by the agonist was abolished by siRNA targeting α7nAChR encoding gene. Consistently, FUT7 knockdown inhibited the adhesive properties of U-937 and prevented them to adhere to endothelial cells. Overexpression of FUT7 also abrogated the adhesion attenuation induced by GTS-21 indicating that FUT7 inhibition was sufficient for inhibiting adhesion by α7nAChR activation. Our work demonstrated that α7nAChR activation regulates monocyte adhesion to endothelial cells through FUT7 inhibition, providing a novel insight into the CAP mechanism., Competing Interests: Declaration of competing interest Tsuyoshi Inoue reports financial support was provided by Kyowa Kirin Co Ltd., (Copyright © 2021 Elsevier Inc. All rights reserved.)
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- 2022
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12. PET imaging of 11 C-labeled thiamine tetrahydrofurfuryl disulfide, vitamin B 1 derivative: First-in-human study.
- Author
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Watanabe Y, Mawatari A, Aita K, Sato Y, Wada Y, Nakaoka T, Onoe K, Yamano E, Akamatsu G, Ohnishi A, Shimizu K, Sasaki M, Doi H, and Senda M
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- 2021
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13. Stability of RNA sequences derived from the coronavirus genome in human cells.
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Wakida H, Kawata K, Yamaji Y, Hattori E, Tsuchiya T, Wada Y, Ozaki H, and Akimitsu N
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- Base Sequence, Betacoronavirus chemistry, COVID-19, Conserved Sequence, Coronaviridae genetics, Genome, Viral, HeLa Cells, Humans, Nucleic Acid Conformation, Pandemics, Severe acute respiratory syndrome-related coronavirus chemistry, SARS-CoV-2, Sequence Analysis, RNA, Betacoronavirus genetics, Coronavirus Infections virology, Pneumonia, Viral virology, RNA Stability, RNA, Viral chemistry, Severe acute respiratory syndrome-related coronavirus genetics
- Abstract
Most viruses inhibit the innate immune system and/or the RNA degradation processes of host cells to construct an advantageous intracellular environment for their survival. Characteristic RNA sequences within RNA virus genomes or RNAs transcribed from DNA virus genomes contribute toward this inhibition. In this study, we developed a method called "Fate-seq" to comprehensively identify the RNA sequences derived from RNA and DNA viruses, contributing RNA stability in the cells. We examined the stabilization activity of 5,924 RNA fragments derived from 26 different viruses (16 RNA viruses and 10 DNA viruses) using next-generation sequencing of these RNAs fused 3' downstream of GFP reporter RNA. With the Fate-seq approach, we detected multiple virus-derived RNA sequences that stabilized GFP reporter RNA, including sequences derived from severe acute respiratory syndrome-related coronavirus (SARS-CoV). Comparative genomic analysis revealed that these RNA sequences and their predicted secondary structures are highly conserved between SARS-CoV and the novel coronavirus, SARS-CoV-2, which is responsible for the global outbreak of the coronavirus-associated disease that emerged in December 2019 (COVID-19). These sequences have the potential to enhance the stability of viral RNA genomes, thereby augmenting viral replication efficiency and virulence., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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14. PET imaging of 11 C-labeled coenzyme Q 10 : Comparison of biodistribution between [ 11 C]ubiquinol-10 and [ 11 C]ubiquinone-10.
- Author
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Watanabe K, Nozaki S, Goto M, Kaneko KI, Hayashinaka E, Irie S, Nishiyama A, Kasai K, Fujii K, Wada Y, Mizuno K, Mizuseki K, Doi H, and Watanabe Y
- Subjects
- Animals, Dietary Supplements analysis, Male, Oxidation-Reduction, Positron-Emission Tomography, Rats, Rats, Sprague-Dawley, Tissue Distribution, Ubiquinone pharmacokinetics, Antioxidants pharmacokinetics, Ubiquinone analogs & derivatives
- Abstract
Coenzyme Q
10 (CoQ10 ) plays a key role not only as an essential electron carrier in the mitochondrial electron transport chain, but also as an antioxidant to protect cells from oxidative stress. CoQ10 supplementation is expected to be effective for a variety of diseases. The predominant forms of CoQ10 are the ubiquinol-10 (reduced form) and ubiquinone-10 (oxidized form). Both forms of CoQ10 supplements are commercially available, however, their kinetic difference is still unclear. In order to conduct in vivo analysis of the kinetics of ubiquinol-10 and ubiquinone-10, we succeeded in synthesizing11 C-labeled ubiquinol-10 ([11 C]UQL) and ubiquinone-10 ([11 C]UQN), respectively. In the present study, we aimed to investigate the kinetics of [11 C]UQL and [11 C]UQN, both of which were administered via the tail vein of 8-week-old male Sprague-Dawley rats. Whole-body positron emission tomography (PET) imaging was performed to follow the time course of accumulation in the liver, spleen, brain, and other organs. Then, at the two typical time points at 20 or 90 min after injection, we conducted the biodistribution study. Various organs/tissues and blood were collected, weighed and counted with a gamma counter. Percent injected dose per gram of tissue (%ID/g) was calculated as the indicator of the accumulation of each compound. As the results, at both time points, %ID/g of [11 C]UQL in the cerebrum, cerebellum, white adipose tissue, muscle, kidney, and testis were higher (P < 0.05) than that of [11 C]UQN: at 90-min time point, %ID/g of [11 C]UQL in the brown adipose tissue was higher (P < 0.05) than that of [11 C]UQN: on the contrary, %ID/g of [11 C]UQL in the spleen was lower (P < 0.05) than that of [11 C]UQN at 90 min. In a separate study of the metabolite analysis in the plasma, UQL injected into the tail vein of rats was almost unchanged during the PET scanning time, but UQN was gradually converted to the reduced form UQL. Therefore, the uptake values of UQL into the tissues and organs were rather accurate but those of UQN might be the sum of UQN uptake and partly converted UQL uptake. These studies suggested that the accumulation level of administered CoQ10 differs depending on its redox state, and that CoQ10 redox state could be crucial for optimization of the effective supplementation., (Copyright © 2019 Elsevier Inc. All rights reserved.)- Published
- 2019
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15. Difference in IgA1 O-glycosylation between IgA deposition donors and IgA nephropathy recipients.
- Author
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Nakazawa S, Imamura R, Kawamura M, Kato T, Abe T, Namba T, Iwatani H, Yamanaka K, Uemura M, Kishikawa H, Nishimura K, Oka K, Tajiri M, Wada Y, and Nonomura N
- Subjects
- Adult, Female, Galactosamine metabolism, Glomerulonephritis, IGA blood, Glomerulonephritis, IGA epidemiology, Glycosylation, Humans, Immunoglobulin A blood, Immunoglobulin A chemistry, Male, Polysaccharides chemistry, Polysaccharides metabolism, Prevalence, Tissue Donors, Glomerulonephritis, IGA immunology, Immunoglobulin A immunology, Kidney Transplantation
- Abstract
IgA nephropathy (IgAN) is the most common form of primary glomerulonephritis, and disease recurrence often occurs after transplantation. On the other hands, Asymptomatic IgA deposition (IgAD) is occasionally observed in donated kidney. It is recognized that IgAD does not progress to IgAN, but the mechanism has not demonstrated yet. In IgAN, aberrant IgA1 O-glycan structure in the hinge region (HR) of serum IgA is suggested as one of the most convincing key mediators. However, little is known about IgA1 O-glycan structure in IgAD patients. Herein, we investigated the prevalence of IgAD in living renal transplant donors in our cohort. IgAD was observed in 21(13.0%) among 161 renal transplant donors and have statistically significant blood relationship with IgAN recipients (28.6% in relatives vs. 9.8% in non-relatives, respectively; p = 0.0073). Next, we evaluated the IgA1 O-glycan structure of serum IgA from IgAN recipients (n = 26), IgAD donors (n = 17), and non-IgAD helthy donors (n = 27) using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). The numbers of GalNAc and Gal and the Gal/GalNAc ratio in the HR of the IgAN recipients had significantly lower comparing to the IgAD and non-IgAD healthy donors. The decreased Gal/GalNAc ratio in IgAN recipients means the increased ratio of galactose-deficient IgA1. To the best of our knowledge, this is the first report to compare the O-glycan structures in IgAN recipients and IgAD donors using MALDI-TOF MS. We concluded that IgAD was more common in IgAN related donors. Overall, decreased GalNAc and Gal contents in HR could play a material pathogenic role in IgAN., (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Published
- 2019
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16. Increased interleukin-1β and basic fibroblast growth factor levels in the cerebrospinal fluid during human herpesvirus-6B (HHV-6B) encephalitis.
- Author
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Tamai M, Kobayashi N, Shimada K, Oka N, Takahashi M, Tanuma A, Tanemoto T, Namba H, Saito Y, Wada Y, Okamoto A, Ida H, and Kondo K
- Subjects
- Astrocytes metabolism, Astrocytes virology, Case-Control Studies, Cell Line, Child, Preschool, DNA, Viral cerebrospinal fluid, Encephalitis, Viral cerebrospinal fluid, Encephalitis, Viral pathology, Encephalitis, Viral virology, Female, Fibroblast Growth Factors cerebrospinal fluid, Gene Expression, Herpesvirus 6, Human growth & development, Herpesvirus 6, Human pathogenicity, Host-Pathogen Interactions, Humans, Infant, Interleukin-1beta cerebrospinal fluid, Male, RNA, Messenger cerebrospinal fluid, RNA, Messenger genetics, Seizures, Febrile cerebrospinal fluid, Seizures, Febrile pathology, Seizures, Febrile virology, DNA, Viral genetics, Encephalitis, Viral genetics, Fibroblast Growth Factors genetics, Herpesvirus 6, Human genetics, Interleukin-1beta genetics, Seizures, Febrile genetics
- Abstract
Human herpesvirus 6B (HHV-6B) causes exanthema subitum in infants and is known to be mildly pathogenic. However, HHV-6B infection can induce febrile seizures in a high percentage of patients, and in rare cases, result in encephalitis. We detected higher levels of interleukin (IL)-1β and basic fibroblast growth factor (bFGF) in the cerebrospinal fluid (CFS) of patients with HHV-6B encephalitis when compared to those in patients with non-HHV-6B-induced febrile seizures. In vitro, IL-1β and bFGF enhanced HHV-6B gene expression in infected U373 astrocytes during the initial and maintenance phases of infection, respectively. These findings indicated that IL-1β and bFGF contribute to HHV-6B growth and the onset of encephalitis., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
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17. Positron emission tomography study on pancreatic somatostatin receptors in normal and diabetic rats with 68Ga-DOTA-octreotide: a potential PET tracer for beta cell mass measurement.
- Author
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Sako T, Hasegawa K, Nishimura M, Kanayama Y, Wada Y, Hayashinaka E, Cui Y, Kataoka Y, Senda M, and Watanabe Y
- Subjects
- Animals, Gallium Radioisotopes analysis, Gallium Radioisotopes metabolism, Male, Octreotide analysis, Octreotide metabolism, Organometallic Compounds metabolism, Pancreas cytology, Radioactive Tracers, Rats, Rats, Sprague-Dawley, Cell Count methods, Diabetes Mellitus pathology, Insulin-Secreting Cells pathology, Octreotide analogs & derivatives, Organometallic Compounds analysis, Pancreas pathology, Positron-Emission Tomography methods, Receptors, Somatostatin analysis
- Abstract
Diabetes mellitus (DM) is a metabolic disorder characterized by hyperglycemia, and the loss or dysfunction of pancreatic beta cells has been reported before the appearance of clinical symptoms and hyperglycemia. To evaluate beta cell mass (BCM) for improving the detection and treatment of DM at earlier stages, we focused on somatostatin receptors that are highly expressed in the pancreatic beta cells, and developed a positron emission tomography (PET) probe derived from octreotide, a metabolically stable somatostatin analog. Octreotide was conjugated with 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), a chelating agent, and labeled with (68)Gallium ((68)Ga). After intravenous injection of (68)Ga-DOTA-octreotide, a 90-min emission scan of the abdomen was performed in normal and DM model rats. The PET studies showed that (68)Ga-DOTA-octreotide radioactivity was highly accumulated in the pancreas of normal rats and that the pancreatic accumulation was significantly reduced in the rats administered with an excess amount of unlabeled octreotide or after treatment with streptozotocin, which was used for the chemical induction of DM in rats. These results were in good agreement with the ex vivo biodistribution data. These results indicated that the pancreatic accumulation of (68)Ga-DOTA-octreotide represented specific binding to the somatostatin receptors and reflected BCM. Therefore, PET imaging with (68)Ga-DOTA-octreotide could be a potential tool for evaluating BCM., (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Published
- 2013
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18. Detection of early stage atherosclerotic plaques using PET and CT fusion imaging targeting P-selectin in low density lipoprotein receptor-deficient mice.
- Author
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Nakamura I, Hasegawa K, Wada Y, Hirase T, Node K, and Watanabe Y
- Subjects
- Animals, Antibodies, Monoclonal, Aorta diagnostic imaging, Aorta metabolism, Aorta pathology, Autoradiography, Biomarkers metabolism, Cholesterol, Dietary administration & dosage, Copper Radioisotopes, Disease Models, Animal, Early Diagnosis, Heterocyclic Compounds, 1-Ring, Mice, Mice, Knockout, Multimodal Imaging, P-Selectin immunology, Plaque, Atherosclerotic etiology, Plaque, Atherosclerotic pathology, Positron-Emission Tomography, Radiopharmaceuticals, Receptors, LDL genetics, Tomography, X-Ray Computed, P-Selectin metabolism, Plaque, Atherosclerotic diagnostic imaging, Plaque, Atherosclerotic metabolism, Receptors, LDL deficiency
- Abstract
Background: Sensitive detection and qualitative analysis of atherosclerotic plaques are in high demand in cardiovascular clinical settings. The leukocyte-endothelial interaction mediated by an adhesion molecule P-selectin participates in arterial wall inflammation and atherosclerosis., Methods and Results: A (64)Cu-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid conjugated anti-P-selectin monoclonal antibody ((64)Cu-DOTA-anti-P-selectin mAb) probe was prepared by conjugating an anti-P-selectin monoclonal antibody with DOTA followed by (64)Cu labeling. Thirty-six hours prior to PET and CT fusion imaging, 3MBq of (64)Cu-DOTA-anti-P-selectin mAb was intravenously injected into low density lipoprotein receptor-deficient Ldlr-/- mice. After a 180min PET scan, autoradiography and biodistribution of (64)Cu-DOTA-anti-P-selectin monoclonal antibody was examined using excised aortas. In Ldlr-/- mice fed with a high cholesterol diet for promotion of atherosclerotic plaque development, PET and CT fusion imaging revealed selective and prominent accumulation of the probe in the aortic root. Autoradiography of aortas that demonstrated probe uptake into atherosclerotic plaques was confirmed by Oil red O staining for lipid droplets. In Ldlr-/- mice fed with a chow diet to develop mild atherosclerotic plaques, probe accumulation was barely detectable in the aortic root on PET and CT fusion imaging. Probe biodistribution in aortas was 6.6-fold higher in Ldlr-/- mice fed with a high cholesterol diet than in those fed with a normal chow diet. (64)Cu-DOTA-anti-P-selectin mAb accumulated selectively in aortic atherosclerotic plaques and was detectable by PET and CT fusion imaging in Ldlr-/- mice., Conclusions: P-selectin is a candidate target molecule for early-phase detection by PET and CT fusion imaging of atherosclerotic plaques., (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Published
- 2013
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19. Quantitative change of IgA hinge O-glycan composition is a novel marker of therapeutic responses of IgA nephropathy.
- Author
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Iwatani H, Inoue T, Wada Y, Nagasawa Y, Yamamoto R, Iijima H, Takehara T, Imai E, Rakugi H, and Isaka Y
- Subjects
- Acetylgalactosamine analysis, Administration, Intravenous, Adolescent, Adrenal Cortex Hormones administration & dosage, Adult, Biomarkers blood, Biomarkers chemistry, Female, Glomerulonephritis, IGA blood, Glomerulonephritis, IGA surgery, Glycosylation, Humans, Immunoglobulin A blood, Male, Polysaccharides analysis, Tonsillectomy, Young Adult, Glomerulonephritis, IGA therapy, Immunoglobulin A chemistry
- Abstract
Aberrant O-glycosylation in the hinge region of serum IgA is suggested to be involved in the pathogenesis of IgA nephropathy (IgAN), because the hypoglycosylation including N-acetylneuraminic acid or galactose has been reported in the mucin-type O-glycan of the hinge portion (HP) of IgA deposited in the IgAN patients' kidney. These aberrant glycosylation has been assessed in most of the previous reports by qualitative but not quantitative methods. In the present study, the molar ratios of GalNAc or Gal to HP were analyzed for serum IgA from IgAN patients. The GalNAc/HP ratio was increased in the patients who achieved remission after a combination therapy of tonsillectomy and intravenous corticosteroid, suggesting any non-innate factors to affect the IgA O-glycosylation in IgAN that is thought to be inherently determined. Furthermore, the O-glycosylation status was different among three groups: IgAN patients in the pretreatment stage, IgAN patients in the remission stage after treatment and healthy controls. These results indicated that aberrant O-glycosylation of serum IgA in the IgAN patients would be inherently present and, to some extent, affected by therapeutic intervention. Finally, the quantitative change of O-glycan composition is a novel marker of therapeutic response of IgAN., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
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20. Brd2 is required for cell cycle exit and neuronal differentiation through the E2F1 pathway in mouse neuroepithelial cells.
- Author
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Tsume M, Kimura-Yoshida C, Mochida K, Shibukawa Y, Amazaki S, Wada Y, Hiramatsu R, Shimokawa K, and Matsuo I
- Subjects
- Alleles, Animals, Cell Cycle genetics, Cell Differentiation, Central Nervous System metabolism, Chromosomal Proteins, Non-Histone, E2F1 Transcription Factor genetics, Mice, Mice, Transgenic, Neurogenesis genetics, Protein Serine-Threonine Kinases genetics, Transcription Factors, Cell Cycle physiology, Central Nervous System growth & development, E2F1 Transcription Factor metabolism, Neuroepithelial Cells cytology, Neurogenesis physiology, Neurons cytology, Protein Serine-Threonine Kinases physiology
- Abstract
To understand genetic programs controlling mammalian central nervous system (CNS) development, we have identified one transgene-inserted mutation, which showed embryonic lethality during neurulation. Determination of the transgene integration site and rescue experiments revealed that the Brd2 gene, whose products specifically bind acetylated histone H4 and can mediate transcription, was the cause of this mutation. Expression studies with specific markers demonstrated that cell cycle progression was accelerated and neuronal differentiation as well as cell cycle exit were impaired in Brd2-deficient neruoepithelial cells. To investigate whether Brd2 regulates neuronal differentiation through a E2F1 transcriptional factor, which directly binds Brd2 and controls genes expression for cell cycle progression and exit, we analyzed Brd2;E2F1 double mutant phenotypes and, consequently found that abnormalities in neuronal differentiation and cell cycle progression due to Brd2-deficiency were restored by removing the E2F1 gene. These findings suggest that Brd2 is required for cell cycle exit and neuronal differentiation of neuroepithelial cells through the E2F1 pathway during mouse CNS development., (Crown Copyright © 2012. Published by Elsevier Inc. All rights reserved.)
- Published
- 2012
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21. B7-DC induced by IL-13 works as a feedback regulator in the effector phase of allergic asthma.
- Author
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Matsumoto K, Fukuyama S, Eguchi-Tsuda M, Nakano T, Matsumoto T, Matsumura M, Moriwaki A, Kan-o K, Wada Y, Yagita H, Shin T, Pardoll DM, Patcharee R, Azuma M, Nakanishi Y, and Inoue H
- Subjects
- Animals, Asthma metabolism, B7-1 Antigen metabolism, Dendritic Cells metabolism, Down-Regulation, Feedback, Physiological, Interferon-gamma genetics, Interferon-gamma immunology, Mice, Mice, Inbred BALB C, Mice, Transgenic, Phenotype, Programmed Cell Death 1 Ligand 2 Protein, Up-Regulation, Asthma immunology, B7-1 Antigen immunology, Hypersensitivity immunology, Interleukin-13 metabolism
- Abstract
B7-DC is a costimulatory molecule belonging to the B7 family. We previously found that treatment with anti-B7-DC mAb during the effector phase enhances asthma phenotypes in mice. We investigated the mechanisms of B7-DC induction and how B7-DC regulates asthma phenotypes. In allergen-challenged IFN-gamma-deficient mice, anti-B7-DC mAb failed to enhance the asthma phenotypes although the induction of B7-DC on dendritic cells of the mice was comparable with that on dendritic cells of wild-type mice. B7-DC on dendritic cells was up-regulated by IL-13 in vitro. The induction of B7-DC on dendritic cells after allergen challenge was attenuated by blockade of IL-13 in vivo. The asthma phenotypes were enhanced in B7-DC-deficient mice, more than in wild-type mice. The enhancement was concurrent with the down-regulation of IFN-gamma and up-regulation of IL-13. These results suggest that B7-DC induced by IL-13 works as a feedback regulator by up-regulating IFN-gamma production during the effector phase of allergic asthma.
- Published
- 2008
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22. Fucoidan induces nitric oxide production via p38 mitogen-activated protein kinase and NF-kappaB-dependent signaling pathways through macrophage scavenger receptors.
- Author
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Nakamura T, Suzuki H, Wada Y, Kodama T, and Doi T
- Subjects
- Animals, HSP90 Heat-Shock Proteins metabolism, Ligands, Macrophages, Peritoneal drug effects, Macrophages, Peritoneal metabolism, Mice, Mice, Knockout, Nitric Oxide Synthase Type II genetics, Nitric Oxide Synthase Type II metabolism, Promoter Regions, Genetic drug effects, Receptors, Scavenger genetics, Signal Transduction, Transcription Factor AP-1 metabolism, Antineoplastic Agents pharmacology, NF-kappa B metabolism, Nitric Oxide biosynthesis, Polysaccharides pharmacology, Receptors, Scavenger drug effects, p38 Mitogen-Activated Protein Kinases metabolism
- Abstract
It has been reported that ligands of the macrophage scavenger receptor (MSR) induce a range of cellular responses including urokinase-type plasminogen activator and the production of inflammatory cytokines. Although nitric oxide (NO) is an important regulatory molecule in physiological functions such as vascular homeostasis, neurotransmission, and host defense, the effect of MSR ligands on NO production from macrophages was unknown. Here, we demonstrate that the MSR ligand, fucoidan, but neither oxidized low-density lipoprotein, acetylated LDL, maleylated bovine serum albumin nor dextran sulfate induces activation of inducible nitric oxide synthase (iNOS) promoter or NO production in RAW264.7 cells. Furthermore, we investigated the molecular mechanism by which fucoidan induces iNOS promoter activation. Using different inhibitors, we showed that the stimulation of fucoidan was mediated by both the p38 mitogen-activated protein kinase and the NF-kappaB-dependent pathways. Although these two pathways were independent, heat shock protein 90 (HSP90) played a significant role in both pathways. Our previous study showed that HSP90 directly interacts with the cytoplasmic domain of MSR. These results provide the evidence that HSP90 bound to the cytoplasmic domain of MSR is implicated in MSR-mediated signal transduction. Moreover, fucoidan-induced NO production by peritoneal macrophages from MSR-knockout (MSR-/-) mice significantly decreases compared with those from wild-type mice. This is the first indication that MSR transduces the signal of fucoidan to iNOS gene expression.
- Published
- 2006
- Full Text
- View/download PDF
23. Tyrosine phosphorylation of clathrin heavy chain under oxidative stress.
- Author
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Ihara Y, Yasuoka C, Kageyama K, Wada Y, and Kondo T
- Subjects
- Animals, Cells, Cultured, Endocytosis physiology, Hydrogen Peroxide pharmacology, Islets of Langerhans cytology, Islets of Langerhans drug effects, Mice, Oxidants pharmacology, Phosphorylation, Pyrimidines pharmacology, Transferrin metabolism, src-Family Kinases antagonists & inhibitors, Clathrin Heavy Chains metabolism, Islets of Langerhans metabolism, Oxidative Stress, Protein Transport physiology, Tyrosine metabolism
- Abstract
In mouse pancreatic insulin-producing betaTC cells, oxidative stress due to H(2)O(2) causes tyrosine phosphorylation in various proteins. To identify proteins bearing phosphotyrosine under stress, the proteins were affinity purified using an anti-phosphotyrosine antibody-conjugated agarose column. A protein of 180kDa was identified as clathrin heavy chain (CHC) by electrophoresis and mass spectrometry. Immunoprecipitated CHC showed tyrosine phosphorylation upon H(2)O(2) treatment and the phosphorylation was suppressed by the Src kinase inhibitor, PP2. The phosphorylation status of CHC affected the intracellular localization of CHC and the clathrin-dependent endocytosis of transferrin under oxidative stress. In conclusion, CHC is a protein that is phosphorylated at tyrosine by H(2)O(2) and this phosphorylation status is implicated in the intracellular localization and functions of CHC under oxidative stress. The present study demonstrates that oxidative stress affects intracellular vesicular trafficking via the alteration of clathrin-dependent vesicular trafficking.
- Published
- 2002
- Full Text
- View/download PDF
24. HSP90, HSP70, and GAPDH directly interact with the cytoplasmic domain of macrophage scavenger receptors.
- Author
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Nakamura T, Hinagata J, Tanaka T, Imanishi T, Wada Y, Kodama T, and Doi T
- Subjects
- Animals, Blotting, Western, COS Cells, Cattle, Chromatography, Affinity, Circular Dichroism, Electrophoresis, Polyacrylamide Gel, Glyceraldehyde-3-Phosphate Dehydrogenases genetics, HSP70 Heat-Shock Proteins genetics, HSP90 Heat-Shock Proteins genetics, Humans, Lung chemistry, Lung metabolism, Peptide Fragments chemical synthesis, Peptide Fragments chemistry, Peptide Fragments metabolism, Precipitin Tests, Protein Binding physiology, Protein Structure, Tertiary physiology, Receptors, Immunologic genetics, Receptors, Scavenger, Scavenger Receptors, Class A, Structure-Activity Relationship, Glyceraldehyde-3-Phosphate Dehydrogenases metabolism, HSP70 Heat-Shock Proteins metabolism, HSP90 Heat-Shock Proteins metabolism, Receptors, Immunologic chemistry, Receptors, Immunologic metabolism
- Abstract
The macrophage scavenger receptor (MSR) is a trimeric membrane protein which binds to modified low-density lipoprotein (LDL) and has been indicated in the development of atherosclerosis. It has recently been demonstrated that the N-terminal cytoplasmic domain of MSR has an important role in the efficient internalization and cell-surface expression of the receptor. This study shows that the N-terminal cytoplasmic domain in bovine was constructed using a peptide architecture technique in which the peptide chain was bundled at their C-terminus to yield a trimeric form and that this did not form an ordered structure. Furthermore, the binding proteins to the cytoplasmic domain of MSR were determined for the first time using a peptide affinity column. Sequence analyses of the specific binding proteins in bovine revealed that heat shock protein 90 (HSP90), heat shock protein 70 (HSP70), leucine aminopeptidase (LAP), adenocylhomocysteinase, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were included. GST-pull-down assay and immunoprecipitation analyses on HSP90, HSP70, and GAPDH showed that all these proteins could bind to the cytoplasmic domain of MSR in vitro and in vivo. These proteins interact with the cytoplasmic domain directly and may have an effect on the functions of MSR such as internalization, cell-surface expression, and signal transduction.
- Published
- 2002
- Full Text
- View/download PDF
25. A new locus for a dominant form of multinodular goiter on 3q26.1-q26.3.
- Author
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Takahashi T, Nozaki J, Komatsu M, Wada Y, Utsunomiya M, Inoue K, Takada G, and Koizumi A
- Subjects
- Adult, Child, Child, Preschool, Chromosome Mapping, Female, Goiter, Nodular pathology, Humans, Male, Pedigree, Thyroid Gland pathology, Chromosomes, Human, Pair 3, Goiter, Nodular genetics
- Abstract
A mass screening program for congenital hypothyroidism has markedly improved prognosis of children with congenital hypothyroidism and also revealed several cases with unknown pathogenesis. We here report two independent Japanese multigeneration families with multinodular goiter (MNG) with euthyroidism and with high TSH. The propositi, 3- and 8-year-old girls in two families, were found during a mass screening. An autosomal dominant pattern of inheritance was suggested in both families. The clinical examinations suggested impaired hormonogenesis but discarded known defects in iodine transport, organification, deficiency of hydrogen peroxide, and thyroid peroxidase. Linkage analysis of the two families including 10 members each using 343 microsatellite markers mapped a single locus independently at D3S1618 (theta = 0) on 3q26.1-q26.3 with a two-point LOD score 3.62 (1.81 for each family) and multipoint LOD score of 3.61 (1.80 for each family). Haplotype inspection delimited an 18-cM interval between D3S1565 and D3S3686., (Copyright 2001 Academic Press.)
- Published
- 2001
- Full Text
- View/download PDF
26. Homozygosity mapping to chromosome 5p15 of a gene responsible for Hartnup disorder.
- Author
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Nozaki J, Dakeishi M, Ohura T, Inoue K, Manabe M, Wada Y, and Koizumi A
- Subjects
- Child, Chromosome Mapping, Chromosomes, Human, Pair 4 genetics, Consanguinity, Female, Genes, Recessive, Genetic Markers, Hartnup Disease diagnosis, Hartnup Disease metabolism, Homozygote, Humans, Japan, Lod Score, Male, Middle Aged, Pedigree, Tryptophan metabolism, Chromosomes, Human, Pair 5 genetics, Genetic Linkage genetics, Hartnup Disease genetics
- Abstract
Hartnup disorder is an autosomal recessive phenotype involving a transporter for monoamino-monocarboxylic acids. Genetic analysis of the mouse model mapped its locus to human chromosome 11q13 (8). We report here the results of linkage analysis in two Japanese first cousin-marriage families. In the first family, the proband had Hartnup disorder and his deceased older brother was reported to have had typical Hartnup symptoms. The younger brother of the proband was shown to have decreased tryptophan absorption by oral loading test. In the second family, a 6-year-old girl, the proband, had specific hyperaminoaciduria. DNA was isolated from either blood samples or umbilical cord stumps. Genome-wide screening by homozygosity mapping was conducted. Taking into account that the older brother was affected and the younger brother was a carrier in the first family, homozygosity mapping (LOD score = 3.55) and GENEHUNTER (LOD score = 3.28) locates the locus of the Hartnup disorder on 5p15., (Copyright 2001 Academic Press.)
- Published
- 2001
- Full Text
- View/download PDF
27. Domain- and site-specific phosphorylation of bovine NF-L by Rho-associated kinase.
- Author
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Hashimoto R, Nakamura Y, Goto H, Wada Y, Sakoda S, Kaibuchi K, Inagaki M, and Takeda M
- Subjects
- Amino Acid Sequence, Animals, Cattle, Guanosine 5'-O-(3-Thiotriphosphate) pharmacology, Intracellular Signaling Peptides and Proteins, Kinetics, Microscopy, Electron, Molecular Sequence Data, Nerve Tissue Proteins chemistry, Nerve Tissue Proteins ultrastructure, Neurofilament Proteins ultrastructure, Peptide Fragments chemistry, Phosphates metabolism, Phosphorylation, Sequence Alignment, Sequence Analysis, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Trypsin metabolism, rho-Associated Kinases, Neurofilament Proteins chemistry, Protein Serine-Threonine Kinases metabolism
- Abstract
Rho-associated kinase (Rho-kinase), the putative target of the small GTP-binding protein Rho, phosphorylated neurofilament protein (NF-L) in vitro with approximately 1 mole phosphate per mole NF-L. Phosphorylated NF-L no longer formed the 10 nm filaments, and NF-L filaments were phosphorylated with a result of nearly complete disassembly. NF-L phosphorylated by Rho-kinase was digested with trypsin, and digested fragments were assigned by MALDI/TOF. Unique phosphorylation sites were found at Ser-26 and Ser-57 in the head domain of NF-L. These results indicate that domain- and site-specific phosphorylation by Rho-kinase may regulate the assembly-disassembly of NF-L filaments.
- Published
- 1998
- Full Text
- View/download PDF
28. The production and purification of functional decorin in a baculovirus system.
- Author
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Gu J, Nakayama Y, Nagai K, and Wada Y
- Subjects
- Animals, Baculoviridae genetics, Cell Line, Chromatography, Ion Exchange, Cloning, Molecular, Collagen antagonists & inhibitors, DNA, Complementary, Decorin, Electrophoresis, Polyacrylamide Gel, Extracellular Matrix Proteins, Humans, Proteoglycans isolation & purification, Proteoglycans metabolism, Spodoptera, Proteoglycans genetics
- Abstract
Human decorin was expressed in Spodoptera frugiperda 21 (Sf21) insect cells. A full-length cDNA encoding preprodecorin of 359 amino acids from a human fibroblast library was cloned into baculovirus transfer vector pVL1392, and transfected into Sf21 insect cells. The infected cells secreted the mature decorin into the culture medium. The secreted decorin lacked glycosaminoglycan but was N-glycosylated, whereas the unmodified decorin was present in the cell lysates, suggesting that N-glycosylation is required for decorin secretion from Sf21 cells. The recombinant decorin was then efficiently purified from the conditioned medium by two chromatographic procedures, hydroxyapatite Sepharose and Con A-Agarose, under nondenaturing conditions. The purified decorin was more potent, as evaluated by the inhibition of collagen fibrillogenesis, than that obtained from bovine tissues under denaturing conditions. The final yield of recombinant decorin was 1.5 mg in 200 ml culture medium of 3 x 10(8) cells. The biologically active decorin produced in Sf21 cells is a potentially useful probe for investigating the molecular interactions of this protein with other extracellular matrix proteins and may also have therapeutic applications.
- Published
- 1997
- Full Text
- View/download PDF
29. Effects of BMP-2, BMP-4, and BMP-6 on osteoblastic differentiation of bone marrow-derived stromal cell lines, ST2 and MC3T3-G2/PA6.
- Author
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Yamaguchi A, Ishizuya T, Kintou N, Wada Y, Katagiri T, Wozney JM, Rosen V, and Yoshiki S
- Subjects
- Alkaline Phosphatase metabolism, Animals, Bone Morphogenetic Proteins, Cell Line, Cyclic AMP biosynthesis, Growth Substances pharmacology, Humans, Mice, Mice, Nude, Parathyroid Hormone pharmacology, Stromal Cells cytology, Stromal Cells transplantation, Bone Marrow Cells, Cell Differentiation, Osteoblasts cytology, Proteins pharmacology
- Abstract
The effects of bone morphogenetic protein-2 (BMP-2) on osteoblastic differentiation of bone marrow stromal cells were investigated using two bone marrow stromal cell lines, ST2 and MC3T3-G2/PA6 (PA6). BMP-2 stimulated ALP activity and induced parathyroid hormone (PTH)-dependent production of cAMP in both ST2 and PA6 cells, but these effects were more apparent in ST2 cells than in PA6 cells. BMP-2 induced the production of osteocalcin in ST2 cells, but not in PA6 cells. BMP-4 and BMP-6 stimulated ALP activity in ST2 cells, but the effect of BMP-6 was less marked than that of BMP-2 and BMP-4. BMP-4 induced PTH-dependent cAMP production of cAMP in ST2 cells, but BMP-6 did not. When ST2 cells were transplanted into the peritoneal cavities of athymic mice with BMP-2 in diffusion chambers, these cells generated mineralized bone in the chambers. These results indicate that BMPs induce the differentiation of bone marrow stromal cells into osteoblasts. However, the effects differ among the BMPs and among the types of cell exposed to these proteins.
- Published
- 1996
- Full Text
- View/download PDF
30. Regulation of fatty acid synthase expression by cholesterol in human cultured cells.
- Author
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Kawabe Y, Sato R, Matsumoto A, Honda M, Wada Y, Yazaki Y, Endo A, Takano T, Itakura H, and Kodama T
- Subjects
- Cell Line, Farnesyl-Diphosphate Farnesyltransferase antagonists & inhibitors, Gene Expression Regulation, Neoplastic drug effects, Hepatoblastoma, Humans, Hydroxycholesterols pharmacology, Kinetics, Liver Neoplasms, RNA, Messenger biosynthesis, Receptors, LDL biosynthesis, Tumor Cells, Cultured, fas Receptor biosynthesis, Bridged Bicyclo Compounds, Heterocyclic pharmacology, Cholesterol physiology, Enzyme Inhibitors pharmacology, Fatty Acid Synthases biosynthesis, Gene Expression Regulation, Enzymologic drug effects, Tricarboxylic Acids pharmacology
- Abstract
The regulation of fatty acid synthase (FAS) expression by sterols in a cultured human hepatoblastoma cell line, Hep G2, was studied. When cells were treated with compactin in a medium containing lipoprotein deficient serum, FAS mRNA level increased 1.6-fold. A squalene synthase inhibitor, TAN1607A, decreased both free and esterified cholesterol contents in Hep G2 cells and increased mRNA levels for FAS, HMG-CoA reductase, squalene synthase and LDL receptor. However, for the increment of FAS mRNA, a 10-fold higher concentration of this inhibitor was needed. These results demonstrate that the concentration of cellular cholesterol which regulates FAS expression is necessarily lower than the levels which regulate other sterol sensitive genes. FAS mRNA was also increased by an inhibitor of SREBP degradation as well as chenodeoxycholic acid. These results indicate that FAS mRNA expression is regulated by cholesterol and is mediated through SREBPs. The implications of the different modes of sterol regulation of FAS and LDL receptor expression are discussed.
- Published
- 1996
- Full Text
- View/download PDF
31. Structure of serum transferrin in carbohydrate-deficient glycoprotein syndrome.
- Author
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Wada Y, Nishikawa A, Okamoto N, Inui K, Tsukamoto H, Okada S, and Taniguchi N
- Subjects
- Child, Child, Preschool, Chromatography, Affinity, Chromatography, High Pressure Liquid, Female, Glycosylation, Humans, Male, Mass Spectrometry, Molecular Weight, Sialic Acids analysis, Syndrome, Transferrin isolation & purification, Carbohydrate Metabolism, Inborn Errors blood, Transferrin chemistry
- Abstract
The structure of the defective transferrin in carbohydrate-deficient glycoprotein syndrome was characterized. Structurally abnormal sugar chains were not found in reversed phase chromatograms of pyridylaminated derivatives from the transferrin of two patients in different families. Electrospray ionization mass spectrometry of the whole transferrin molecules revealed an abnormal species that was smaller than normal tetrasialotransferrin by 2,200 daltons, just the size of the disialylated biantennary sugar chain. These data indicated that the disialotransferrin specifically found in this syndrome is missing either of two N-linked sugar chains, suggesting a metabolic error in the early steps of protein glycosylation.
- Published
- 1992
- Full Text
- View/download PDF
32. Distinct clustering of point mutations in mitochondrial DNA among patients with mitochondrial encephalomyopathies and with Parkinson's disease.
- Author
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Ozawa T, Tanaka M, Ino H, Ohno K, Sano T, Wada Y, Yoneda M, Tanno Y, Miyatake T, and Tanaka T
- Subjects
- Adolescent, Adult, Female, Humans, Male, Middle Aged, Phylogeny, DNA, Mitochondrial chemistry, Mitochondria, Muscle pathology, Multigene Family, Mutation, Parkinson Disease genetics
- Abstract
The total sequence data for mitochondrial DNA (mtDNA) revealed distinct clustering of point mutations (pms) in mtDNA among one patient with myoclonus epilepsy with ragged-red fibers (MERRF), two patients with Parkinson's disease (PD), two patients with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS), and one patient with fatal infantile cardiomyopathy (FICM). Among 33 to 62 pms found in each patients, sequentially diverged five clusters of pms were detected and designated as C-1 to C-5. C-1, consisted of fourteen pms, existed in the MERRF patient, C-1 and C-2 (nine pms) in one PD patient, C-1 to C-3 (seven pms) in another PD patient, C-1 to C-4 (one pm) in one MELAS patient and C-1 to C-5 (three pms) in another MELAS patient and the FICM patient. From these clustering of pms, a phylogenetic tree of mitochondrial encephalomyopathies (ME) was constructed. This tree clearly indicated that the ME and PD patients are members of the same gene family, and the MELAS and FICM patients are each others' closest relative. Each patient's unique pms (14 to 28 pms) were detected and, from their characteristic features, the types of the mutations specific for the disease were classified as mit- + syn- for MERRF, mit- + p- for PD, and syn- + mit- for MELAS. An inverse relation was found between the total number of pms and life span of the MELAS and FICM patients.
- Published
- 1991
- Full Text
- View/download PDF
33. Mitochondrial DNA mutations in mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS).
- Author
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Tanaka M, Ino H, Ohno K, Ohbayashi T, Ikebe S, Sano T, Ichiki T, Kobayashi M, Wada Y, and Ozawa T
- Subjects
- Adolescent, Amino Acid Sequence, Base Composition, Base Sequence, Female, Humans, Male, Molecular Sequence Data, NAD(P)H Dehydrogenase (Quinone), Nucleic Acid Conformation, Oligonucleotide Probes, Quinone Reductases deficiency, RNA, Transfer, Leu genetics, Syndrome, DNA, Mitochondrial genetics, Metabolism, Inborn Errors genetics, Mitochondria, Muscle enzymology, Mutation, Quinone Reductases genetics
- Abstract
The total sequences of mitochondrial DNA were determined in two patients with juvenile-onset mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) due to Complex I deficiency. Patients 1 and 2 had three and two unique point mutations, respectively, causing replacement of phylogenically conserved amino acids. A transition from G to A was found at nucleotide position 5601 in the alanine tRNA gene of Patient 2, and a transition from A to G was found at 3243 in the leucine (UUR) tRNA gene of both patients. The latter mutation located at the phylogenically conserved 5' end of the dihydrouridine loop of the tRNA molecule, and was present in two patients with adult-onset MELAS and absent in controls. These results indicate that a mass of mtDNA mutations including the A-to-G transition in the tRNA(Leu) gene is a genetic cause of MELAS.
- Published
- 1991
- Full Text
- View/download PDF
34. Demonstration of a new mouse mammary tumor virus locus in the genome of the mammary tumor prone strain SHN mouse.
- Author
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Koizumi A, Tsukada M, Wada Y, and Kamiyama S
- Subjects
- Animals, Blotting, Southern, DNA isolation & purification, DNA Probes, DNA, Neoplasm isolation & purification, DNA, Viral isolation & purification, Deoxyribonuclease EcoRI, Disease Susceptibility, Female, Mammary Neoplasms, Experimental microbiology, Mammary Tumor Virus, Mouse genetics, Mice, Mice, Inbred Strains, Plasmids, Restriction Mapping, DNA genetics, DNA, Neoplasm genetics, DNA, Viral genetics, Mammary Neoplasms, Experimental genetics, Mammary Tumor Virus, Mouse isolation & purification
- Abstract
Until this report, there have been no detailed analyses of the mouse mammary tumor virus (MMTV) loci in the mammary tumor prone strain SHN. Using a probe, which hybridizes to the env sequence of MMTV, Southern blotting of genomic DNA from the brain after digestion with EcoRI revealed 5 endogenous proviruses: Mtv-1 (4.5kb), Mtv-2 (11 kb), Mtv-8 (6.7kb), Mtv-17 (8.3kb) and a newly-found 6.5-kb fragment. F1-hybrid mice (C3H/He female x SHN male) also possessed the 6.5-kb fragment. Thus, we conclude that the 6.5-kb fragment is unique to SHN mice. Genomic DNA from mammary tumors of SHN mice showed MMTV insertions, suggesting that activation of an oncogene(s) occurred in this strain.
- Published
- 1990
- Full Text
- View/download PDF
35. Cloning and sequence of cDNA encoding a peptide C-terminal alpha-amidating enzyme from Xenopus laevis.
- Author
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Mizuno K, Ohsuye K, Wada Y, Fuchimura K, Tanaka S, and Matsuo H
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Molecular Sequence Data, Skin enzymology, Xenopus laevis, Cloning, Molecular, DNA isolation & purification, Mixed Function Oxygenases, Multienzyme Complexes, Oxygenases genetics
- Abstract
The C-terminal alpha-amide formation of the peptides is one of the most important events of prohormone processing. We have recently isolated an alpha-amidating enzyme, AE-I, from Xenopus laevis skin, which is the only enzyme ever purified to homogeneity. In this study, we report cloning and sequence of cDNA encoding AE-I. Our results indicate that enzyme AE-I is initially synthesized as a precursor with 400 amino acid residues, which is further processed to the mature enzyme consisting of 344 residues. Preliminary expression in E. coli of the cDNA corresponding to AE-I was found to produce an enzyme with appreciable alpha-amidating activity.
- Published
- 1987
- Full Text
- View/download PDF
36. Cloning of cDNA encoding a new peptide C-terminal alpha-amidating enzyme having a putative membrane-spanning domain from Xenopus laevis skin.
- Author
-
Ohsuye K, Kitano K, Wada Y, Fuchimura K, Tanaka S, Mizuno K, and Matsuo H
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, DNA Restriction Enzymes, DNA, Recombinant, Molecular Sequence Data, Nucleic Acid Hybridization, Oxidoreductases Acting on CH-NH Group Donors metabolism, RNA, Messenger genetics, Sequence Homology, Nucleic Acid, Skin analysis, Xenopus laevis, Cell Membrane metabolism, DNA genetics, Deoxyribonucleases, Type II Site-Specific, Enzyme Precursors genetics, Mixed Function Oxygenases, Multienzyme Complexes, Oxidoreductases Acting on CH-NH Group Donors genetics
- Abstract
A cDNA clone encoding a precursor of a peptide C-terminal alpha-amidating enzyme (AE-I) from Xenopus laevis skin was recently isolated and sequenced in our laboratory. In this study, by using the restriction fragment of this clone as a hybridization probe, we have identified the cDNA encoding another new peptide C-terminal alpha-amidating enzyme (tentatively named AE-II) distinct from AE-I. The cDNA encodes a polypeptide of 875 amino acid residues, which contains a region extensively homologous to AE-I precursor at N-terminus. The encoded protein characteristically has a putative membrane-spanning domain near C-terminus. Our results indicate that C-terminal alpha-amide formation of peptides in Xenopus skin is regulated by at least two distinct alpha-amidating enzymes.
- Published
- 1988
- Full Text
- View/download PDF
37. NAD (P) H-dependent reduction of nicotinamide N-oxide by an unique enzyme system consisting of liver microsomal NADPH-cytochrome C reductase and cytosolic aldehyde oxidase.
- Author
-
Kitamura S, Wada Y, and Tatsumi K
- Subjects
- Aldehyde Oxidase, Animals, Cytosol enzymology, Electron Transport, Male, NAD metabolism, NADP metabolism, Niacinamide metabolism, Rabbits, Aldehyde Oxidoreductases metabolism, Microsomes, Liver enzymology, NADPH-Ferrihemoprotein Reductase metabolism, Niacinamide analogs & derivatives
- Abstract
NAD (P) H-dependent reduction of nicotinamide N-oxide was investigated with rabbit liver preparations. Microsomes, microsomal NADPH-cytochrome c reductase or cytosolic aldehyde oxidase alone exhibited no nicotinamide N-oxide reductase activity in the presence of NADPH or NADH. However, when the microsomal preparations were combined with the cytosolic enzyme, a significant N-oxide reductase activity was observed in the presence of the reduced pyridine nucleotide. The activity was enhanced by FAD or methyl viologen. Cytosol alone supplemented with NADPH or NADH exhibited only a slight, but when combined with microsomes, a significant N-oxide reductase activity. Based on these facts, we propose a new electron transfer system consisting of NADPH-cytochrome c reductase and aldehyde oxidase, which exhibits nicotinamide N-oxide reductase activity in the presence of the reduced pyridine nucleotide.
- Published
- 1984
- Full Text
- View/download PDF
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