154 results on '"Yukiko, Yoshida"'
Search Results
2. Sugar-Recognizing Ubiquitin Ligases: Action Mechanisms and Physiology
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Yukiko Yoshida, Tsunehiro Mizushima, and Keiji Tanaka
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E3 ubiquitin ligase ,ERAD ,F-box protein ,glycoprotein ,N-glycan ,SCF complex ,Physiology ,QP1-981 - Abstract
F-box proteins, the substrate recognition subunits of SKP1–CUL1–F-box protein (SCF) E3 ubiquitin ligase complexes, play crucial roles in various cellular events mediated by ubiquitination. Several sugar-recognizing F-box proteins exist in both mammalian and plant cells. Although glycoproteins generally reside outside of cells, or in organelles of the secretory pathway, these lectin-type F-box proteins reside in the nucleocytoplasmic compartment. Mammalian sugar-recognizing F-box proteins commonly bind to the innermost position of N-glycans through a unique small hydrophobic pocket in their loops. Two cytosolic F-box proteins, Fbs1 and Fbs2, recognize high-mannose glycans synthesized in the ER, and SCFFbs1 and SCFFbs2 ubiquitinate excess unassembled or misfolded glycoproteins in the ERAD pathway by recognizing the innermost glycans, which serve as signals for aberrant proteins. On the other hand, endomembrane-bound Fbs3 recognizes complex glycans as well as high-mannose glycans, and SCFFbs3 ubiquitinates exposed glycoproteins in damaged lysosomes fated for elimination by selective autophagy. Plants express stress-inducible lectin-type F-box proteins recognizing a wider range of N- and O-glycans, suggesting that the roles of mammalian and plant lectin-type F-box proteins have diverged over the course of evolution to recognize species-specific targets with distinct functions. These sugar-recognizing F-box proteins interpret glycans in the cytosol as markers of unwanted proteins and organelles, and degrade them via the proteasome or autophagy.
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- 2019
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3. Freshly isolated retinal capillaries to determine efflux transporter function at the inner BRB
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Kosuke, Tajima, Shin-Ichi, Akanuma, Yuki, Ohishi, Yukiko, Yoshida, Björn, Bauer, Yoshiyuki, Kubo, Masahiko, Inouye, and Ken-Ichi, Hosoya
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ATP Binding Cassette Transporter, Subfamily G, Member 2 ,Animals ,Pharmaceutical Science ,Biological Transport ,ATP Binding Cassette Transporter, Subfamily B, Member 1 ,Multidrug Resistance-Associated Proteins ,Rats, Wistar ,Retina ,Capillaries ,Neoplasm Proteins ,Rats - Abstract
Since it has been known that in vitro cell lines for analyzing drug transport at the inner blood-retinal barrier (BRB) do not completely retain several in vivo functions, new ex vivo/in vitro methods to evaluate drug transport across the inner BRB help us understand the role of this barrier in maintaining the homeostasis of vision and regulating drug distribution to the retina. To expand the limitations of existing in vitro approaches, we established a protocol to isolate fresh rat retinal capillaries as ex vivo model of the inner BRB. Fresh retinal capillaries were prepared by applying serial filtration steps and using density gradient centrifugation. We performed mRNA and protein analyses by reverse transcription-polymerase chain reaction and immunostaining that indicated expression of marker proteins such as facilitative glucose transporter 1 and claudin-5 in freshly isolated rat retinal capillaries. We also used fluorescent transporter substrates to characterize functional activity of organic anion transporter (Oat) 3, P-glycoprotein (P-gp), breast cancer resistance protein (Bcrp), and multidrug resistance-associated protein (Mrp) 4 in isolated retinal capillaries. Capillary luminal accumulation of fluorescent substrates of P-glycoprotein and Bcrp was decreased in the presence of transporter inhibitors. Moreover, luminal accumulation of the Oat3 and Mrp4 substrate, 8-(2-[fluoresceinyl]aminoethylthio) adenosine-3',5'-cyclic monophosphate (8-[fluo]-cAMP), was reduced by substrates/inhibitors of Oat3 and Mrp4. In conclusion, our study shows that freshly isolated retinal capillaries retain marker protein expression and transporter functional activity. It is suggested that isolated retinal capillaries are a useful tool to study transport across the inner BRB. Using freshly isolated retinal capillaries, we anticipate applying this approach to determine the role of transporters at the inner BRB during pathophysiological states of the eye and evaluate the drug delivery to the retina.
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- 2022
4. Luminescence detection of peptide:N-glycanase activity using engineered split inteins
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Tsuyoshi Takahashi, Tatsuya Uchibayashi, Nozomi Ishii, Ichiro Matsuo, Yukiko Yoshida, and Tadashi Suzuki
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Materials Chemistry ,Metals and Alloys ,Ceramics and Composites ,General Chemistry ,Catalysis ,Surfaces, Coatings and Films ,Electronic, Optical and Magnetic Materials - Abstract
A convenient method to detect peptide:N-glycanase activity was developed using engineered split inteins in which the N-intein sequences contain N,N′-diacetylchitobiose.
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- 2022
5. Ever-expanding NGLY1 biology
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Tadashi, Suzuki and Yukiko, Yoshida
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Congenital Disorders of Glycosylation ,Glycosylation ,Humans ,Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase ,Endoplasmic Reticulum-Associated Degradation ,General Medicine ,Biology ,Molecular Biology ,Biochemistry - Abstract
The cytosolic peptide:N-glycanase (PNGase; NGLY1 in humans) is a deglycosylating enzyme that is widely conserved in eukaryotes. This enzyme is involved in the degradation of misfolded N-glycoproteins that are destined for proteasomal degradation in the cytosol, a process that is called endoplasmic reticulum-associated degradation. Although the physiological significance of NGLY1 remained unknown until recently, the discovery of NGLY1 deficiency, a human genetic disorder bearing mutations in the NGLY1 gene, has led to explosive research progress regarding the functional characterization of this enzyme. For example, it is now known that NGLY1 can also act as an ‘editing enzyme’ to convert N-glycosylated asparagine residues to aspartate residues, thus introducing negative charges into a core peptide and modulating the function of the target molecule. Diverse biological processes have also been found to be affected by compromised NGLY1 activity. In this special issue, recent research progress on the functional characterization of NGLY1 and its orthologues in worm/fly/rodents, assay methods/biomarkers useful for the development of therapeutics and the comprehensive transcriptome/proteome of NGLY1-KO cells as well as patient-derived cells are discussed.
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- 2021
6. Mining Interesting Patterns Using Estimated Frequencies from Subpatterns and Superpatterns.
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Yukiko Yoshida, Yuiko Ohta, Kenichi Kobayashi 0001, and Nobuhiro Yugami
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- 2003
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7. Inflammation-Induced Attenuation of Prostaglandin D2 Elimination across Rat Blood–Brain Barrier: Involvement of the Downregulation of Organic Anion Transporter 3 and Multidrug Resistance-Associated Protein 4
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Kahori Hashimoto, Yukiko Yoshida, Ken Ichi Hosoya, Shin Ichi Akanuma, and Yoshiyuki Kubo
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0301 basic medicine ,Organic anion transporter 1 ,Lipopolysaccharide ,Pharmaceutical Science ,Prostaglandin ,Inflammation ,Pharmacology ,Blood–brain barrier ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Downregulation and upregulation ,medicine ,Neuroinflammation ,integumentary system ,biology ,Chemistry ,General Medicine ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,biology.protein ,lipids (amino acids, peptides, and proteins) ,Prostaglandin D2 ,medicine.symptom - Abstract
Prostaglandin (PG) D2 is a lipid mediator, and in the brain, overproduction of PGD2 is reportedly involved in the progression and exacerbation of neuroinflammation. The objective of this study was to elucidate PGD2 efflux transport, under normal and inflammatory conditions, across the blood-brain barrier (BBB), which is formed by brain capillaries. Elimination of [3H]PGD2 across the BBB of normal and lipopolysaccharide (LPS)-induced inflammatory rats was examined by the intracerebral microinjection technique. After intracerebral injection, the percentage of [3H]PGD2 remaining in the ipsilateral cerebrum decreased with time, with a half-life of 13 min. This [3H]PGD2 elimination across the BBB was significantly inhibited by the co-administration of unlabeled PGD2, which suggests carrier-mediated PGD2 efflux transport at the BBB. In isolated rat brain capillaries, mRNA expression of organic anion transporter (Oat) 3, organic anion-transporting polypeptide (Oatp) 1a4, and multidrug resistance-associated protein (Mrp) 4 was observed. In addition, co-administration of substrates/inhibitors for Oat3, Oatp1a4, and/or Mrp4, such as benzylpenicillin and cefmetazole, reduced [3H]PGD2 elimination across the BBB. Data suggest that Oat3 and Mrp4, but not Oatp1a4 are involved in PGD2 elimination across the BBB, as Oatp1a4-expressing Xenopus (X.) oocytes did not show the significant [3H]PGD2 uptake compared with water-injected X. oocytes. In LPS-treated rats, [3H]PGD2 elimination across the BBB and mRNA expression levels of Oat3 and Mrp4 were significantly decreased. Our data suggest that Oat3- and Mrp4-mediated PGD2 elimination across the BBB is attenuated under inflammatory conditions.
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- 2020
8. Gaview - A Visualization Tool for Supporting GA Simulations and Analyses.
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Yukiko Yoshida, Nobue Adachi, and Koji Tajima
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- 1996
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9. A Diploid Genetic Algorithm for Preserving Population Diversity - pseudo-Meiosis GA.
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Yukiko Yoshida and Nobue Adachi
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- 1994
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10. Loss of peptide
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Yukiko, Yoshida, Makoto, Asahina, Arisa, Murakami, Junko, Kawawaki, Meari, Yoshida, Reiko, Fujinawa, Kazuhiro, Iwai, Ryuichi, Tozawa, Noriyuki, Matsuda, Keiji, Tanaka, and Tadashi, Suzuki
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Cell Nucleus ,Mice, Knockout ,Proteasome Endopeptidase Complex ,SKP Cullin F-Box Protein Ligases ,Behavior, Animal ,Cell Death ,Nuclear Respiratory Factor 1 ,Ubiquitination ,Motor Activity ,Biological Sciences ,HCT116 Cells ,Models, Biological ,Mice, Inbred C57BL ,Protein Transport ,Cytosol ,Polysaccharides ,Mutation ,Animals ,Humans ,Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase ,Sugars ,Cell Proliferation ,HeLa Cells - Abstract
Mutations in the human peptide:N-glycanase gene (NGLY1), which encodes a cytosolic de–N-glycosylating enzyme, cause a congenital autosomal recessive disorder. In rodents, the loss of Ngly1 results in severe developmental delay or lethality, but the underlying mechanism remains unknown. In this study, we found that deletion of Fbxo6 (also known as Fbs2), which encodes a ubiquitin ligase subunit that recognizes glycoproteins, rescued the lethality-related defects in Ngly1-KO mice. In NGLY1-KO cells, FBS2 overexpression resulted in the substantial inhibition of proteasome activity, causing cytotoxicity. Nuclear factor, erythroid 2–like 1 (NFE2L1, also known as NRF1), an endoplasmic reticulum–associated transcriptional factor involved in expression of proteasome subunits, was also abnormally ubiquitinated by SCF(FBS2) in NGLY1-KO cells, resulting in its retention in the cytosol. However, the cytotoxicity caused by FBS2 was restored by the overexpression of “glycan-less” NRF1 mutants, regardless of their transcriptional activity, or by the deletion of NRF1 in NGLY1-KO cells. We conclude that the proteasome dysfunction caused by the accumulation of N-glycoproteins, primarily NRF1, ubiquitinated by SCF(FBS2) accounts for the pathogenesis resulting from NGLY1 deficiency.
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- 2021
11. Loss of peptide: N -glycanase causes proteasome dysfunction mediated by a sugar-recognizing ubiquitin ligase
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Arisa Murakami, Tadashi Suzuki, Noriyuki Matsuda, Kazuhiro Iwai, Reiko Fujinawa, Yukiko Yoshida, Keiji Tanaka, Ryuichi Tozawa, Makoto Asahina, Meari Yoshida, and Junko Kawawaki
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0303 health sciences ,Multidisciplinary ,biology ,Chemistry ,Endoplasmic reticulum ,Endoplasmic-reticulum-associated protein degradation ,NFE2L1 ,Ubiquitin ligase ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,Ubiquitin ,Proteasome ,biology.protein ,NRF1 ,NGLY1 ,030217 neurology & neurosurgery ,030304 developmental biology - Abstract
Significance Cytosolic peptide: N -glycanase (NGLY1) is a widely conserved enzyme involved in de– N -glycosylation of N -glycosylated proteins. Mutations in the human NGLY1 gene cause global developmental delay and multisystemic symptoms, but the molecular mechanism underlying pathogenesis remains poorly understood. FBXO6/FBS2, a subunit of the SCF (SKP1–CUL1–F-box protein) ubiquitin ligase complex, recognizes N -glycans of cytosolic glycoproteins in the endoplasmic reticulum–associated degradation (ERAD) pathway. This paper reports that high levels of ERAD glycoprotein substrates abnormally ubiquitinated by SCF FBS2 in the absence of NGLY1 impair the proteasome, contributing to the pathogenesis of NGLY1 deficiency. Importantly, knockout of Fbxo6/Fbs2 rescued the lethality of NGLY1 deficiency in mice, suggesting a strategy for developing therapeutics for this intractable disease.
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- 2021
12. The Structural Differences between a Glycoprotein Specific F-Box Protein Fbs1 and Its Homologous Protein FBG3.
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Taichi Kumanomidou, Kazuya Nishio, Kenji Takagi, Tomomi Nakagawa, Atsuo Suzuki, Takashi Yamane, Fuminori Tokunaga, Kazuhiro Iwai, Arisa Murakami, Yukiko Yoshida, Keiji Tanaka, and Tsunehiro Mizushima
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Medicine ,Science - Abstract
The Skp1-Cul1-F-box protein (SCF) complex catalyzes protein ubiquitination in diverse cellular processes and is one of the best-characterized ubiquitin ligases. F-box proteins determine the substrate specificities of SCF ubiquitin ligases. Among these, Fbs1/FBG1/FBXO2, Fbs2/FBG2/FBXO6, and Fbs3/FBG5/FBXO27 recognize the N-glycans of glycoproteins, whereas FBG3/FBXO44 has no sugar-binding activity, despite the high sequence homology and conservation of the residues necessary for oligosaccharide binding between Fbs1-3 and FBG3. Here we determined the crystal structure of the Skp1-FBG3 complex at a resolution of 2.6 Å. The substrate-binding domain of FBG3 is composed of a 10-stranded antiparallel β-sandwich with three helices. Although the overall structure of FBG3 is similar to that of Fbs1, the residues that form the Fbs1 carbohydrate-binding pocket failed to be superposed with the corresponding residues of FBG3. Structure-based mutational analysis shows that distinct hydrogen bond networks of four FBG3 loops, i.e., β2-β3, β5-β6, β7-β8, and β9-β10, prevent the formation of the carbohydrate-binding pocket shown in Fbs1.
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- 2015
- Full Text
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13. Inflammation-Induced Attenuation of Prostaglandin D
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Shin-Ichi, Akanuma, Kahori, Hashimoto, Yukiko, Yoshida, Yoshiyuki, Kubo, and Ken-Ichi, Hosoya
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Inflammation ,Lipopolysaccharides ,Male ,Brain Diseases ,Microinjections ,Prostaglandin D2 ,Cefmetazole ,Down-Regulation ,Penicillin G ,Organic Anion Transporters, Sodium-Independent ,Rats ,Disease Models, Animal ,Xenopus laevis ,Blood-Brain Barrier ,Oocytes ,Animals ,Humans ,Multidrug Resistance-Associated Proteins - Abstract
Prostaglandin (PG) D
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- 2020
14. A substrate-trapping strategy to find E3 ubiquitin ligase substrates identifies Parkin and TRIM28 targets
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Yasushi Saeki, Fumiaki Ohtake, Takeshi Kondo, Hiroaki Yaguchi, Masashi Watanabe, Yusuke Kasuga, Shigetsugu Hatakeyama, Hiroki Ishida, Hidehisa Takahashi, Masanobu Suzuki, Yukiko Yoshida, and Keiji Tanaka
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Proteomics ,TRIM28 ,Ubiquitylation ,Ubiquitin-Protein Ligases ,Proteomic analysis ,Medicine (miscellaneous) ,Plasma protein binding ,Tripartite Motif-Containing Protein 28 ,Article ,General Biochemistry, Genetics and Molecular Biology ,Parkin ,Substrate Specificity ,03 medical and health sciences ,0302 clinical medicine ,Ubiquitin ,Protein Interaction Mapping ,Gene Knockdown Techniques ,Humans ,Protein Interaction Domains and Motifs ,lcsh:QH301-705.5 ,030304 developmental biology ,0303 health sciences ,biology ,Protein Stability ,Chemistry ,Cell Cycle ,Ubiquitination ,Reproducibility of Results ,Proteases ,Cell biology ,Ubiquitin ligase ,lcsh:Biology (General) ,030220 oncology & carcinogenesis ,biology.protein ,General Agricultural and Biological Sciences ,Transcription factor II B ,Cyclin A2 ,Protein Binding - Abstract
The identification of true substrates of an E3 ligase is biologically important but biochemically difficult. In recent years, several techniques for identifying substrates have been developed, but these approaches cannot exclude indirect ubiquitination or have other limitations. Here we develop an E3 ligase substrate-trapping strategy by fusing a tandem ubiquitin-binding entity (TUBE) with an anti-ubiquitin remnant antibody to effectively identify ubiquitinated substrates. We apply this method to one of the RBR-type ligases, Parkin, and to one of the RING-type ligases, TRIM28, and identify previously unknown substrates for TRIM28 including cyclin A2 and TFIIB. Furthermore, we find that TRIM28 promotes cyclin A2 ubiquitination and degradation at the G1/S phase and suppresses premature entry into S phase. Taken together, the results indicate that this method is a powerful tool for comprehensively identifying substrates of E3 ligases., Watanabe et al. combine two previously developed strategies to identify E3 ubiquitin ligase substrates into a method, TR-TUBE that is subsequently used to identify substrates of the Parkin and TRIM28 ligases. They identify known substrates, validating the utility of the approach, and find that TRIM28 targets Cyclin A and TFIIB for degradation.
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- 2020
15. Participation of Monocarboxylate Transporter 8, But Not P-Glycoprotein, in Carrier-Mediated Cerebral Elimination of Phenytoin across the Blood-Brain Barrier
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Ken Ichi Hosoya, Ryuta Jomura, Björn Bauer, Yoshiyuki Kubo, Shin-Ichi Akanuma, and Yukiko Yoshida
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Phenytoin ,Male ,Monocarboxylic Acid Transporters ,ATP Binding Cassette Transporter, Subfamily B ,Pharmaceutical Science ,Mice, Transgenic ,02 engineering and technology ,Pharmacology ,Blood–brain barrier ,030226 pharmacology & pharmacy ,Article ,03 medical and health sciences ,Mice ,0302 clinical medicine ,In vivo ,medicine ,otorhinolaryngologic diseases ,Animals ,Pharmacology (medical) ,heterocyclic compounds ,P-glycoprotein ,Monocarboxylate transporter ,biology ,Symporters ,Chemistry ,Organic Chemistry ,digestive, oral, and skin physiology ,Transporter ,Isolated brain ,021001 nanoscience & nanotechnology ,nervous system diseases ,Rats ,stomatognathic diseases ,medicine.anatomical_structure ,Blood-Brain Barrier ,Models, Animal ,biology.protein ,Molecular Medicine ,Anticonvulsants ,Female ,Efflux ,0210 nano-technology ,Biotechnology ,medicine.drug ,Half-Life - Abstract
PURPOSE: In this study, we investigated in detail the transport of phenytoin across the blood-brain barrier (BBB) to identify the transporter(s) involved in BBB-mediated phenytoin efflux from the brain. METHODS: We evaluated the brain-to-blood efflux transport of phenytoin in vivo by determining the brain efflux index (BEI) and uptake in brain slices. We additionally conducted brain perfusion experiments and BEI studies in P-glycoprotein (P-gp)-deficient mice. In addition, we determined the mRNA expression of monocarboxylate transporter (MCT) in isolated brain capillaries and performed phenytoin uptake studies in MCT-expressing Xenopus oocytes. RESULTS: [(14)C]Phenytoin brain efflux was time-dependent with a half-life of 17 min in rats and 31 min in mice. Intracerebral pre-administration of unlabeled phenytoin attenuated BBB-mediated phenytoin efflux transport, suggesting carrier-mediated phenytoin efflux transport across the BBB. Pre-administration of P-gp substrates in rats and genetic P-gp deficiency in mice did not affect BBB-mediated phenytoin efflux transport. In contrast, pre-administration of MCT8 inhibitors attenuated phenytoin efflux. Moreover, rat MCT8-expressing Xenopus oocytes exhibited [(14)C]phenytoin uptake, which was inhibited by unlabeled phenytoin. CONCLUSION: Our data suggest that MCT8 at the BBB participates in phenytoin efflux transport from the brain to the blood.
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- 2020
16. Lectin-Type Ubiquitin Ligase Subunits: Fbs Proteins and Their Applications for Use
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Yukiko, Yoshida
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F-Box Proteins ,Ubiquitination ,Cell Cycle Proteins ,Nerve Tissue Proteins ,Protein Engineering ,Recombinant Proteins ,Substrate Specificity ,Cytosol ,Polysaccharides ,Mutation ,Humans ,Mannose ,Glycoproteins ,HeLa Cells - Abstract
Three lectin-type F-box proteins called Fbs (F-box protein-recognizing sugar chains) are found in mammals, and function as substrate-binding subunits in the SCF (Skp1/Cullin1/F-box protein) complex ubiquitin ligases. The SCF
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- 2020
17. Lectin-Type Ubiquitin Ligase Subunits: Fbs Proteins and Their Applications for Use
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Yukiko Yoshida
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chemistry.chemical_classification ,0303 health sciences ,Glycan ,biology ,Chemistry ,F-box protein ,Cell biology ,Ubiquitin ligase ,03 medical and health sciences ,Cytosol ,0302 clinical medicine ,Ubiquitin ,Organelle ,Skp1 ,biology.protein ,Glycoprotein ,030217 neurology & neurosurgery ,030304 developmental biology - Abstract
Three lectin-type F-box proteins called Fbs (F-box protein-recognizing sugar chains) are found in mammals, and function as substrate-binding subunits in the SCF (Skp1/Cullin1/F-box protein) complex ubiquitin ligases. The SCFFbs recognizes cytosolic N-glycans as a signal for an adverse cellular state, and ubiquitinates glycoproteins which appear in the cytosol to remove them from cells. Although Fbs proteins recognize innermost Man3GlcNAc2 structure that is commonly found in most N-glycan structures, they preferentially bind high-mannose-type glycans. Recently, the recombinant Fbs1 derivative protein has been developed as a tool for comprehensive enrichment of N-glycopeptides. The labeled Fbs3 is also available as a tool for detecting organelle damage in cells as it has characteristic properties which cause it to quickly accumulate in damaged organelles. In this chapter, we introduce two applications of use for Fbs proteins: the unbiased N-glycopeptide capture method and the detection of damaged organelles in living cells.
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- 2020
18. The Generation of Human γδT Cell-Derived Induced Pluripotent Stem Cells from Whole Peripheral Blood Mononuclear Cell Culture
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Daisuke Watanabe, Yukiko Yoshida, Mariko Taniguchi-Ikeda, Takashi Aoi, Takeshi Azuma, and Michiyo Koyanagi-Aoi
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Pluripotent Stem Cells ,0301 basic medicine ,Cell- and Tissue-Based Therapy ,gamma-delta TCR ,T‐lymphocytes ,Biology ,Lymphocyte Activation ,Sendai virus ,Zoledronic Acid ,Peripheral blood mononuclear cell ,03 medical and health sciences ,0302 clinical medicine ,Translational Research Articles and Reviews ,gamma‐delta TCR ,Neoplasms ,Humans ,Cytotoxic T cell ,Progenitor cell ,Induced pluripotent stem cell ,Intraepithelial Lymphocytes ,Cells, Cultured ,T-lymphocytes ,Gene Transfer Techniques ,Receptors, Antigen, T-Cell, gamma-delta ,Reprogramming ,Cell Biology ,General Medicine ,Cell sorting ,Cellular Reprogramming ,Cell biology ,Induced pluripotent stem cells ,Haematopoiesis ,030104 developmental biology ,Hematopoietic Stem/Progenitor Cells ,030220 oncology & carcinogenesis ,Interleukin-2 ,immunotherapy ,Stem cell ,T-Lymphocytes, Cytotoxic ,Developmental Biology - Abstract
γδT cells constitute a small proportion of lymphocytes in peripheral blood. Unlike αβT cells, the anti-tumor activities are exerted through several different pathways in a MHC-unrestricted manner. Thus, immunotherapy using γδT cells is considered to be effective for various types of cancer. Occasionally, however, ex vivo expanded cells are not as effective as expected due to cell exhaustion. To overcome the issue of T-cell exhaustion, researchers have generated induced pluripotent stem cells (iPSCs) that harbor the same T-cell receptor (TCR) genes as their original T-cells, which provide nearly limitless sources for antigen-specific cytotoxic T lymphocytes (CTLs). However, these technologies have focused on αβT cells and require a population of antigen-specific CTLs, which are purified by cell sorting with HLA-peptide multimer, as the origin of iPS cells. In the present study, we aimed to develop an efficient and convenient system for generating iPSCs that harbor rearrangements of the TCRG and TCRD gene regions (γδT-iPSCs) without cell-sorting. We stimulated human whole peripheral blood mononuclear cell (PBMC) culture using Interleukin-2 and Zoledronate to activate γδT cells. Gene transfer into those cells with the Sendai virus vector resulted in γδT cell-dominant expression of exogenous genes. The introduction of reprogramming factors into the stimulated PBMC culture allowed us to establish iPSC lines. Around 70% of the established lines carried rearrangements at the TCRG and TCRD gene locus. The γδT-iPSCs could differentiate into hematopoietic progenitors. Our technology will pave the way for new avenues toward novel immunotherapy that can be applied for various types of cancer.
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- 2017
19. Analysis of the impact of regional temperature pattern on the energy consumption in the commercial sector in Japan
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Yujiro Hirano, Yukiko Yoshida, Tsuyoshi Fujita, Kei Gomi, Daisuke Narumi, and Shogo Nakamura
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Offset (computer science) ,Meteorology ,Mechanical Engineering ,0208 environmental biotechnology ,02 engineering and technology ,Building and Construction ,Estimating equations ,Energy consumption ,010501 environmental sciences ,01 natural sciences ,020801 environmental engineering ,Climatology ,Environmental science ,Electrical and Electronic Engineering ,0105 earth and related environmental sciences ,Civil and Structural Engineering - Abstract
We conducted this study to clarify the impact of regional climatic conditions on the energy consumption in the commercial sector, in Japan. For this research, we utilized the Database for the Energy Consumption of Commercial Buildings (DECC), which provided data on the individual buildings. The existing energy consumption pattern derived from the individual building data seems to have large variations depending on climatic conditions. Therefore, in this research, connections were made between the seasonal fluctuations in energy consumption as well as the seasonal temperature changes, in order to extract the components of variation due to temperature, and make a regional comparison. In addition, we prepared estimating equations for the energy consumption based on temperature. By applying the regional temperature data to these equations, the impact of temperature alone was extracted for comparison. The results showed that, for the types of buildings addressed in this research, even though there is a regionality to the ratio of increments during summer and winter with regard to heating and cooling, it is offset by summing them up. In this research, we were able to roughly determine the level of fluctuations in the energy consumption as an impact of temperature.
- Published
- 2017
20. Ubiquitination of exposed glycoproteins by SCF FBXO27 directs damaged lysosomes for autophagy
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Sayaka Yasuda, Toshiharu Fujita, Keiji Tanaka, Yukiko Yoshida, Yasushi Saeki, Maho Hamasaki, Tamotsu Yoshimori, Arisa Murakami, Junko Kawawaki, Noriyuki Matsuda, and Kazuhiro Iwai
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0301 basic medicine ,VAMP3 ,chemistry.chemical_classification ,Multidisciplinary ,LAMP2 ,biology ,LAMP1 ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,medicine.anatomical_structure ,Ubiquitin ,chemistry ,Biochemistry ,Lysosome ,Ubiquitin ligase complex ,medicine ,biology.protein ,CUL1 ,Glycoprotein - Abstract
Ubiquitination functions as a signal to recruit autophagic machinery to damaged organelles and induce their clearance. Here, we report the characterization of FBXO27, a glycoprotein-specific F-box protein that is part of the SCF (SKP1/CUL1/F-box protein) ubiquitin ligase complex, and demonstrate that SCFFBXO27 ubiquitinates glycoproteins in damaged lysosomes to regulate autophagic machinery recruitment. Unlike F-box proteins in other SCF complexes, FBXO27 is subject to N-myristoylation, which localizes it to membranes, allowing it to accumulate rapidly around damaged lysosomes. We also screened for proteins that are ubiquitinated upon lysosomal damage, and identified two SNARE proteins, VAMP3 and VAMP7, and five lysosomal proteins, LAMP1, LAMP2, GNS, PSAP, and TMEM192. Ubiquitination of all glycoproteins identified in this screen increased upon FBXO27 overexpression. We found that the lysosomal protein LAMP2, which is ubiquitinated preferentially on lysosomal damage, enhances autophagic machinery recruitment to damaged lysosomes. Thus, we propose that SCFFBXO27 ubiquitinates glycoproteins exposed upon lysosomal damage to induce lysophagy.
- Published
- 2017
21. Strategies for a sustainable campus in Osaka University
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Yoshiyuki Shimoda, Takumi Ohashi, and Yukiko Yoshida
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Engineering ,business.industry ,Energy management ,020209 energy ,Mechanical Engineering ,02 engineering and technology ,Building and Construction ,010501 environmental sciences ,Environmental economics ,01 natural sciences ,Energy engineering ,Renewable energy ,Outsourcing ,Unit (housing) ,Energy conservation ,Fiscal year ,0202 electrical engineering, electronic engineering, information engineering ,Operations management ,Electrical and Electronic Engineering ,business ,0105 earth and related environmental sciences ,Civil and Structural Engineering ,Efficient energy use - Abstract
This paper reports strategies toward achieving a sustainable campus at Osaka University, Japan. We advance a daily energy-use schedule for each building type to provide effective energy-saving strategies. We classify the facilities into three categories according to department type to reveal their energy-use patterns and identify their strategies for saving energy. These strategies will contribute to a sustainable campus using photovoltaic generation. Not only the implementation of energy-saving technologies but also the installation of renewable energy technologies will realize a sustainable campus at Osaka University. Category I (liberal arts) buildings have low energy density. The energy efficiency strategy for Category I is to develop zero-energy buildings by tuning the energy system to people's daily routines. Category II (science and engineering) buildings have high energy density. The energy efficiency strategy for Category II is effective energy conservation using the plan–do–check–act cycle. Category III buildings are large-scale facilities with experimental laboratories. The energy efficiency strategy for Category III is outsourcing energy management to an energy service company. Compared with fiscal year 2010, in 2015 Osaka University decreased energy use per unit floor space by 22%. It achieved these savings despite an increase in total floor area during that period.
- Published
- 2017
22. Assessing the effects of CO2 reduction strategies on heat islands in urban areas
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Yujiro Hirano and Yukiko Yoshida
- Subjects
Renewable Energy, Sustainability and the Environment ,business.industry ,020209 energy ,Geography, Planning and Development ,Photovoltaic system ,Environmental engineering ,Climate change ,Transportation ,02 engineering and technology ,Building design ,Thermal insulation ,Environmental protection ,Greenhouse gas ,0202 electrical engineering, electronic engineering, information engineering ,Environmental science ,Urban heat island ,business ,Roof ,Built environment ,Civil and Structural Engineering - Abstract
There has been a wide range of low-carbon solutions proposed to mitigate climate change. However, such measures must be compatible with the local environment and living standards of residents to be brought to fruition. Measures that adversely affect residential environments will be difficult to implement, so the impacts of measures on the local environment must be taken into consideration during implementation. This study assessed the effects on urban heat islands of efforts to reduce CO2 emissions, as one environmental impact associated with climate change. A simulated assessment was conducted, using an urban canopy model coupled with a building energy model (CM-BEM), to evaluate the effects of five specific measures: solar shading of windows using curtains and blinds, improvement of the thermal insulation of building walls and roof surfaces, implementation of energy-saving measures related to indoor appliances, installation of solar photovoltaic (PV) panels, and adjustment of preset cooling temperatures. The study focused on these effects as they occur within typical urban districts of office buildings, fire-resistant housing, and wooden housing. Results indicated that many of the energy-saving measures have slight temperature lowering effects, but solar panel installation and improved heat insulation, both associated with changes in surface heat balances, tend to raise daytime temperatures to some extent. However, effects on daytime temperatures were in the range of 0.1–0.2 °C and, as such, none of the CO2 reduction measures considered was deemed a significant factor in raising urban temperatures.
- Published
- 2016
23. Detection of ubiquitination activity and identification of ubiquitinated substrates using TR-TUBE
- Author
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Yukiko, Yoshida, Yasushi, Saeki, Hikaru, Tsuchiya, and Keiji, Tanaka
- Subjects
Deubiquitinating Enzymes ,Ubiquitin ,Ubiquitin-Protein Ligases ,Ubiquitination ,Animals ,Humans ,Trypsin ,Polyubiquitin ,Ubiquitinated Proteins ,Enzyme Assays ,Substrate Specificity - Abstract
Ubiquitination is a transient posttranslational modification; polyubiquitin chains are removed from proteins by deubiquitinating enzymes (DUBs) and many ubiquitinated proteins are degraded by the proteasome. Exogenously expressed trypsin-resistant tandem ubiquitin-binding entity (TR-TUBE) protects polyubiquitin chains from DUBs and inhibits proteasomal degradation in cells. TR-TUBE effectively binds to substrates ubiquitinated by an exogenously expressed ubiquitin ligase, and enables detection of the specific activity of a given ubiquitin ligase and isolation of its substrates. In this chapter, we describe methods for the detection of ubiquitin ligase activity as well as the identification of substrates of a given ubiquitin ligase using two enrichment tools, TR-TUBE and anti-diGly antibody, coupled with mass spectrometry (MS).
- Published
- 2019
24. Sugar-Recognizing Ubiquitin Ligases: Action Mechanisms and Physiology
- Author
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Keiji Tanaka, Tsunehiro Mizushima, and Yukiko Yoshida
- Subjects
glycoprotein ,0301 basic medicine ,F-box protein ,Physiology ,Mini Review ,SCF complex ,Endoplasmic-reticulum-associated protein degradation ,sugar chain ,lcsh:Physiology ,03 medical and health sciences ,0302 clinical medicine ,Physiology (medical) ,ERAD pathway ,Secretory pathway ,chemistry.chemical_classification ,lcsh:QP1-981 ,biology ,ERAD ,Ubiquitin ligase ,Cell biology ,030104 developmental biology ,E3 ubiquitin ligase ,Proteasome ,chemistry ,N-glycan ,biology.protein ,Glycoprotein ,030217 neurology & neurosurgery - Abstract
F-box proteins, the substrate recognition subunits of SKP1–CUL1–F-box protein (SCF) E3 ubiquitin ligase complexes, play crucial roles in various cellular events mediated by ubiquitination. Several sugar-recognizing F-box proteins exist in both mammalian and plant cells. Although glycoproteins generally reside outside of cells, or in organelles of the secretory pathway, these lectin-type F-box proteins reside in the nucleocytoplasmic compartment. Mammalian sugar-recognizing F-box proteins commonly bind to the innermost position of N-glycans through a unique small hydrophobic pocket in their loops. Two cytosolic F-box proteins, Fbs1 and Fbs2, recognize high-mannose glycans synthesized in the ER, and SCFFbs1 and SCFFbs2 ubiquitinate excess unassembled or misfolded glycoproteins in the ERAD pathway by recognizing the innermost glycans, which serve as signals for aberrant proteins. On the other hand, endomembrane-bound Fbs3 recognizes complex glycans as well as high-mannose glycans, and SCFFbs3 ubiquitinates exposed glycoproteins in damaged lysosomes fated for elimination by selective autophagy. Plants express stress-inducible lectin-type F-box proteins recognizing a wider range of N- and O-glycans, suggesting that the roles of mammalian and plant lectin-type F-box proteins have diverged over the course of evolution to recognize species-specific targets with distinct functions. These sugar-recognizing F-box proteins interpret glycans in the cytosol as markers of unwanted proteins and organelles, and degrade them via the proteasome or autophagy.
- Published
- 2019
25. Detection of ubiquitination activity and identification of ubiquitinated substrates using TR-TUBE
- Author
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Hikaru Tsuchiya, Yukiko Yoshida, Keiji Tanaka, and Yasushi Saeki
- Subjects
biology ,Proteasome ,Ubiquitin ,Ubiquitinated Proteins ,Ubiquitin ligase activity ,Chemistry ,biology.protein ,Posttranslational modification ,Specific activity ,Ubiquitin ligase ,Cell biology ,Deubiquitinating enzyme - Abstract
Ubiquitination is a transient posttranslational modification; polyubiquitin chains are removed from proteins by deubiquitinating enzymes (DUBs) and many ubiquitinated proteins are degraded by the proteasome. Exogenously expressed trypsin-resistant tandem ubiquitin-binding entity (TR-TUBE) protects polyubiquitin chains from DUBs and inhibits proteasomal degradation in cells. TR-TUBE effectively binds to substrates ubiquitinated by an exogenously expressed ubiquitin ligase, and enables detection of the specific activity of a given ubiquitin ligase and isolation of its substrates. In this chapter, we describe methods for the detection of ubiquitin ligase activity as well as the identification of substrates of a given ubiquitin ligase using two enrichment tools, TR-TUBE and anti-diGly antibody, coupled with mass spectrometry (MS).
- Published
- 2019
26. Loss of peptide:N-glycanase causes proteasome dysfunction mediated by a sugar-recognizing ubiquitin ligase.
- Author
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Yukiko Yoshida, Makoto Asahina, Arisa Murakami, Junko Kawawaki, Meari Yoshida, Reiko Fujinawa, Kazuhiro Iwai, Ryuichi Tozawa, Noriyuki Matsuda, Keiji Tanaka, and Tadashi Suzuki
- Subjects
- *
UBIQUITIN , *GLYCOPROTEINS , *DEVELOPMENTAL delay , *HUMAN genes , *PATHOGENESIS , *COMMERCIAL products - Abstract
Mutations in the human peptide:N-glycanase gene (NGLY1), which encodes a cytosolic de-N-glycosylating enzyme, cause a congenital autosomal recessive disorder. In rodents, the loss of Ngly1 results in severe developmental delay or lethality, but the underlying mechanism remains unknown. In this study, we found that deletion of Fbxo6 (also known as Fbs2), which encodes a ubiquitin ligase subunit that recognizes glycoproteins, rescued the lethality-related defects in Ngly1-KO mice. In NGLY1-KO cells, FBS2 overexpression resulted in the substantial inhibition of proteasome activity, causing cytotoxicity. Nuclear factor, erythroid 2-like 1 (NFE2L1, also known as NRF1), an endoplasmic reticulum-associated transcriptional factor involved in expression of proteasome subunits, was also abnormally ubiquitinated by SCFFBS2 in NGLY1-KO cells, resulting in its retention in the cytosol. However, the cytotoxicity caused by FBS2 was restored by the overexpression of "glycan-less" NRF1 mutants, regardless of their transcriptional activity, or by the deletion of NRF1 in NGLY1-KO cells. We conclude that the proteasome dysfunction caused by the accumulation of N-glycoproteins, primarily NRF1, ubiquitinated by SCFFBS2 accounts for the pathogenesis resulting from NGLY1 deficiency. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
27. Structural analysis of a function-associated loop mutant of the substrate-recognition domain of Fbs1 ubiquitin ligase
- Author
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Kazuya Nishio, Tsunehiro Mizushima, Yukiko Yoshida, and Keiji Tanaka
- Subjects
Models, Molecular ,Protein Conformation, alpha-Helical ,0301 basic medicine ,RBX1 ,Mutant ,Biophysics ,Gene Expression ,Cell Cycle Proteins ,Nerve Tissue Proteins ,Crystallography, X-Ray ,Biochemistry ,F-box protein ,Research Communications ,Substrate Specificity ,Structure-Activity Relationship ,03 medical and health sciences ,0302 clinical medicine ,Structural Biology ,Skp1 ,Escherichia coli ,Genetics ,Humans ,Protein Interaction Domains and Motifs ,Amino Acid Sequence ,Cloning, Molecular ,chemistry.chemical_classification ,DNA ligase ,Binding Sites ,SKP Cullin F-Box Protein Ligases ,Sequence Homology, Amino Acid ,biology ,Chemistry ,F-Box Proteins ,Endoplasmic reticulum ,Hydrogen Bonding ,Condensed Matter Physics ,Recombinant Proteins ,Ubiquitin ligase ,Cell biology ,030104 developmental biology ,030220 oncology & carcinogenesis ,Mutation ,biology.protein ,Protein Conformation, beta-Strand ,CUL1 ,Sequence Alignment ,Plasmids ,Protein Binding - Abstract
The SCF ubiquitin ligase comprises four components: Skp1, Cul1, Rbx1 and a variable-subunit F-box protein. The F-box protein Fbs1, which recognizes the N-linked glycoproteins, is involved in the endoplasmic reticulum-associated degradation pathway. Although FBG3, another F-box protein, shares 51% sequence identity with Fbs1, FBG3 does not bind glycoproteins. To investigate the sequence–structure relationship of the substrate-binding pocket, the crystal structure of a mutant substrate-binding domain of Fbs1 in which the six nonconserved regions (β1, β2–β3, β3–β4, β5–β6, β7–β8 and β9–β10) of Fbs1 were substituted with those of FBG3 was determined. The substrate-binding pocket of this model exhibits structural features that differ from those of Fsb1.
- Published
- 2016
28. Estimating residential CO2 emissions based on daily activities and consideration of methods to reduce emissions
- Author
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Yujiro Hirano, Yukiko Yoshida, and Tomohiko Ihara
- Subjects
Consumption (economics) ,Environmental Engineering ,Natural resource economics ,020209 energy ,Geography, Planning and Development ,Environmental engineering ,Sample (statistics) ,02 engineering and technology ,Building and Construction ,Energy consumption ,Family income ,Environmental studies ,Work (electrical) ,Secondary sector of the economy ,0202 electrical engineering, electronic engineering, information engineering ,Production (economics) ,Business ,Civil and Structural Engineering - Abstract
To achieve a low-carbon society, it is crucial to make the transition away from wasteful lifestyles that are shaped by mass production/mass consumption towards low-carbon lifestyles. Research in the field of urban environmental studies examining human behavioral factors affecting energy consumption, has mostly focused on CO 2 emissions relating to direct energy consumption, such as the use of air conditioners and water heaters. As CO 2 emissions from the industrial sector are greater than those from the residential and transportation sectors in Japan, it would be prudent for consumers to work on reducing CO 2 emissions relating to products and services. In an effort to provide some basic research to achieve the aforementioned objectives, this study looked at statistical data from sources such as the Japanese Family Income and Expenditure Survey and the Japanese National Survey of Family Income and Expenditure to estimate direct and indirect CO 2 emissions relating to daily expenditures and consumption. As a case study, we selected a sample denizen to calculate the CO 2 emissions relating to daily living. Based on the obtained results, we examined the lifestyle and consumption patterns and their relation to CO 2 emissions. The results suggest that while it is still important for consumers to reduce their direct energy consumption in an effort to create a low-carbon society, there is still potential for indirect CO 2 emission reductions in areas of production and consumption.
- Published
- 2016
29. Phenobarbital reduces blood glucose and gluconeogenesis through down-regulation of phosphoenolpyruvate carboxykinase (GTP) gene expression in rats
- Author
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Hiroaki Oda, Yukiko Yoshida, Noriko Kimura, Atsushi Kakinuma, and Yuji Okuda
- Subjects
Blood Glucose ,Male ,endocrine system ,medicine.medical_specialty ,endocrine system diseases ,GTP' ,Biophysics ,Down-Regulation ,Biology ,digestive system ,Biochemistry ,Gene Expression Regulation, Enzymologic ,Diabetes Mellitus, Experimental ,Tyrosine aminotransferase ,Downregulation and upregulation ,Internal medicine ,Gene expression ,medicine ,Animals ,Homeostasis ,Humans ,Insulin ,Rats, Wistar ,Molecular Biology ,Gene Expression Profiling ,Gluconeogenesis ,nutritional and metabolic diseases ,Hep G2 Cells ,Cell Biology ,Rats ,Glucose ,medicine.anatomical_structure ,Endocrinology ,Phenobarbital ,Hepatocyte ,Hepatocytes ,Phosphoenolpyruvate Carboxykinase (GTP) ,Phosphoenolpyruvate carboxykinase ,hormones, hormone substitutes, and hormone antagonists - Abstract
The regulatory mechanism of phosphoenolpyruvate carboykinase (GTP) (EC 4.1.1.32) (PEPCK) gene expression and gluconeogenesis by phenobarbital (PB), which is known to induce drug-metabolizing enzymes, was investigated. Higher level of PEPCK mRNA was observed in spherical rat primary hepatocytes on EHS-gel than monolayer hepatocytes on TIC (type I collagen). We found that PB directly suppressed PEPCK gene expression in spherical hepatocytes on EHS-gel, but not in those on TIC. PB strongly suppressed cAMP-dependent induction of PEPCK gene expression. Tyrosine aminotransferase (TAT), another gluconeogenic enzyme, was induced by cAMP, but not suppressed by PB. Chronic administration of PB reduced hepatic PEPCK mRNA in streptozotocin-induced diabetic and nondiabetic rats, and PB reduced blood glucose level in diabetic rats. Increased TAT mRNA in diabetic rats was not suppressed by PB. These results indicated that PB-dependent reduction is specific to PEPCK. From pyrvate challenge test, PB suppressed the increased gluconeogenesis in diabetic rats. PEPCK gene promoter activity was suppressed by PB in HepG2 cells. In conclusion, we found that spherical hepatocytes cultured on EHS-gel are capable to respond to PB to suppress PEPCK gene expression. Moreover, our results indicate that hypoglycemic action of PB result from transcriptional repression of PEPCK gene and subsequent suppression of gluconeogenesis.
- Published
- 2015
30. High expression of FOXM1 critical for sustaining cell proliferation in mitochondrial DNA-less liver cancer cells
- Author
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Ken-ichi Fujita, Motoko Shibanuma, Kazunori Mori, Tsuyoshi Maruyama, Yusuke Nogami, Yukiko Yoshida, Masato Higurashi, and Fumihiro Ishikawa
- Subjects
0301 basic medicine ,Mitochondrial DNA ,Carcinoma, Hepatocellular ,DNA Copy Number Variations ,Cell ,Biology ,DNA, Mitochondrial ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,medicine ,Humans ,Gene ,Transcription factor ,Cells, Cultured ,Cell Proliferation ,Cell growth ,Forkhead Box Protein M1 ,Liver Neoplasms ,Cell Biology ,Up-Regulation ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,medicine.anatomical_structure ,Cell culture ,030220 oncology & carcinogenesis ,Cancer cell ,Hepatocytes ,FOXM1 ,Cancer research - Abstract
The copy number of mitochondrial DNA (mtDNA) is decreased in most cancer types, including hepatocellular carcinoma (HCC), compared to normal counterparts. However, a decrease in mtDNA usually leads to defects in cell proliferation, which contradicts the robustness of cancer cell proliferation. In this study, we found that four out of seven HCC cell lines were of the mtDNA-less type. Interestingly, FOXM1, a member of the FOX transcription factor family, was highly expressed in a subset of them with proliferative potential maintained. B-MYB, a partner of FOXM1, was also expressed in the same cell lines. RNAi-mediated experiments demonstrated that when FOXM1/B-MYB was silenced in the cell lines, cell cycle-related genes were downregulated, while p21Cip1 was induced with senescence-associated β-galactosidase, resulting in G1/S cell cycle arrest. These results suggest that high expression of FOXM1/B-MYB is critical for sustaining cell proliferation in mtDNA-less cells. In addition, we found that high expression of FOXM1 was mediated by the deubiquitinating enzyme, OTUB1, in one cell line. Thus, interference with FOXM1/B-MYB expression, such as through OTUB1 inhibition, may induce a dormant state of senescence-like proliferation arrest in mtDNA-less cancer cells. This finding may be utilized for the development of precision medicine for relevant cancers.
- Published
- 2020
31. A diploid genetic algorithm for preserving population diversity — Pseudo-Meiosis GA
- Author
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Yukiko, Yoshida, primary and Nobue, Adachi, additional
- Published
- 1994
- Full Text
- View/download PDF
32. Assessment of urine partial oxygen pressure to predict postoperative acute kidney injury in major surgical patients.
- Author
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Naoto IKEMOTO, Hiroshi KATAYAMA, Naomasa ISHIDA, Masaaki MICHIDA, Yukiko YOSHIDA, Yoko OCHIAI, Masao HAYASHI, Munenori TAKAOKA, Tomoki YAMATSUJI, Ichiro OHASHI, and Hideki NAKATSUKA
- Published
- 2021
- Full Text
- View/download PDF
33. Ubiquitination of exposed glycoproteins by SCF
- Author
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Yukiko, Yoshida, Sayaka, Yasuda, Toshiharu, Fujita, Maho, Hamasaki, Arisa, Murakami, Junko, Kawawaki, Kazuhiro, Iwai, Yasushi, Saeki, Tamotsu, Yoshimori, Noriyuki, Matsuda, and Keiji, Tanaka
- Subjects
SKP Cullin F-Box Protein Ligases ,Autophagy ,Ubiquitination ,Humans ,Biological Sciences ,Lysosomes ,Glycoproteins ,HeLa Cells - Abstract
Although lysosomes play a crucial role in autophagy, damaged lysosomes are eliminated by autophagy. The molecular mechanisms that recognize lysosomal damage in cells remain poorly understood, but ubiquitination is a known prerequisite for directing autophagic machinery to damaged lysosomes. FBXO27, a substrate-recognition subunit of the SCF (SKP1/CUL1/F-box protein) ubiquitin ligase complex, localizes to the cytosolic surface of endomembranes and binds glycoproteins. This paper reports that SCFFBXO27 ubiquitinates exposed glycoproteins normally sequestered on the lumenal surface of lysosomes following lysosomal damage, resulting in accelerated recruitment of autophagic machinery.
- Published
- 2017
34. Case of non-mosaic trisomy 20 in amniotic fluid cultures without anomalies in the fetus: Cytogenetic discrepancy between amniocytes and fetal blood
- Author
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Minoru Irahara, Soichiro Nakayama, Issei Imoto, Kazuhisa Maeda, Yukiko Yoshida, and Takashi Kaji
- Subjects
Gynecology ,medicine.medical_specialty ,Fetus ,Amniotic fluid ,medicine.diagnostic_test ,business.industry ,Obstetrics and Gynecology ,Gestational age ,Prenatal diagnosis ,Karyotype ,Andrology ,medicine.anatomical_structure ,Placenta ,medicine ,Amniocentesis ,Confined placental mosaicism ,business - Abstract
We report a case of non-mosaic trisomy 20 detected prenatally by amniocentesis during the 16th week of pregnancy. Fetal blood sampling showed a normal karyotype and no fetal, neonatal or infant abnormalities were observed. Amniotic fluid cell karyotyping revealed a trisomy 20 (47,XY,+20) with 100% trisomic cells (38/38); however, a subsequent cordocentesis revealed a normal male karyotype. Moreover, a follow-up ultrasonographic examination did not reveal any major congenital malformations, and a healthy male infant was delivered subsequently at an appropriate gestational age without obvious anomalies. Cytogenetic analysis of blood lymphocytes from the infant revealed a normal karyotype, but cultured cells from the term placenta showed a mosaic karyotype 47,XY,+20/46,XY with 88% trisomic cells (44 of 50). Furthermore, no anomalies or developmental delays were observed in the neonatal period, thus suggesting two possibilities: confined placental mosaicism with the presence of normal and abnormal cell lineages, or generalized mosaicism affecting a limited number of tissues in both the placenta and fetus.
- Published
- 2014
35. Multinuclear and Ground Glass-Formed Cells Detected in the Peritoneal Dialysate
- Author
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Naoki Washida, Matsuhiko Hayashi, Kentaro Fujii, Kaori Kameyama, Hirobumi Tokuyama, Yukiko Yoshida, Shu Wakino, Ayumi Yoshifuji, Yujiro Machida, Hiroshi Itoh, and Takahiro Kasai
- Subjects
Microbiology (medical) ,Pathology ,medicine.medical_specialty ,business.industry ,medicine.medical_treatment ,Immunosuppression ,medicine.disease_cause ,medicine.disease ,Peritoneal dialysate ,Peritoneal dialysis ,03 medical and health sciences ,0302 clinical medicine ,Infectious Diseases ,Herpes simplex virus ,030220 oncology & carcinogenesis ,medicine ,030211 gastroenterology & hepatology ,Bullous pemphigoid ,business - Published
- 2018
36. Occupational burnout and work factors in community and hospital midwives: A survey analysis
- Author
-
Yukiko Yoshida and Jane Sandall
- Subjects
Adult ,Male ,Working hours ,medicine.medical_specialty ,Nurse Midwives ,media_common.quotation_subject ,Protective factor ,Stress recognition ,Workload ,Burnout ,Midwifery ,Occupational burnout ,Young Adult ,Nursing ,Residence Characteristics ,Surveys and Questionnaires ,Maternity and Midwifery ,Humans ,Medicine ,Statistical analysis ,Burnout, Professional ,media_common ,business.industry ,Obstetrics and Gynecology ,Middle Aged ,Health Surveys ,Hospitals ,England ,Work (electrical) ,Family medicine ,Female ,business ,psychological phenomena and processes ,Autonomy - Abstract
Background: community-based midwifery practice has been promoted in the UK maternity policy over the last decade as a means of increasing continuity of care. However, there have been growing concerns to suggest that the community-based continuity model may not be sustainable due to the high levels of occupational burnout in midwives resulted by increased on-call work. Aims: this paper attempted to identify work factors associated with the levels of burnout in community midwives as compared to hospital midwives, aiming at contributing to the debate of organising sustainable midwifery care. Methods: a statistical analysis was conducted drawing on data from a survey of all midwives working at one Hospital Trust in England (n¼ 238). Occupational burnout was measured using the Maslach Burnout Inventory (MBI). Results: the sample midwives (n¼ 128, 54%) had significantly higher levels of burnout compared to the reference groups. Multiple regression analysis identified as follows: (1) high levels of occupational autonomy were a key protective factor of burnout, and more prevalent in the community, (2) working hours were positively associated with burnout, and community midwives were more likely to have higher levels of stress recognition, and (3) support for work-life-balance from the Trust had a significant protective effect on the levels of burnout. Conclusion: the results should be taken into account in the maternity policy in order to incorporate continuity of care and sustainable organisation of midwifery care.
- Published
- 2013
37. LED Lighting and Energy Saving in Nagoya University(<Special Issue>Tokai Branch, Create the New Technology Fits the Occasion with the Succession of Arts)
- Author
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Satoru Kuno and Yukiko Yoshida
- Subjects
LED lamp ,Architectural engineering ,Engineering ,Low energy ,law ,business.industry ,Ecological succession ,Electrical and Electronic Engineering ,business ,The arts ,Simulation ,Energy (signal processing) ,law.invention - Published
- 2013
38. [Development of a method for identifying the substrates of specific ubiquitin ligases]
- Author
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Yukiko, Yoshida
- Subjects
Ubiquitin ,Ubiquitin-Protein Ligases ,Humans ,Substrate Specificity - Published
- 2016
39. Cloning and sequencing of the 3' half of a potato virus Y (O strain) genome encoding the 5k protein, protease, polymerase and coat protein.
- Author
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Makoto Hidaka, Yukiko Yoshida, Haruhiko Masaki, Shigetou Namba, Shuichi Yamashita, Tsuneo Tsuchizaki, and Takeshi Uozumi
- Published
- 1992
- Full Text
- View/download PDF
40. STUDY ON THE REDUCTION OF COOLING LOAD IN MODERATE SEASONS USING NATURAL VENTILATION AT OFFICE BUILDINGS
- Author
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Yukiko Yoshida
- Subjects
Reduction (complexity) ,Environmental Engineering ,Waste management ,Cooling load ,Environmental science ,Natural ventilation ,Urban heat island - Published
- 2011
41. DEVELOPMENT OF ENERGY MONITORING SYSTEM FOR RESIDENTS
- Author
-
Yukiko Yoshida
- Subjects
Architectural engineering ,Development (topology) ,Computer science ,Energy management ,Architecture ,Systems engineering ,Monitoring system ,Building and Construction ,Energy (signal processing) - Published
- 2011
42. SCFFbx2-E3-ligase-mediated degradation of BACE1 attenuates Alzheimer’s disease amyloidosis and improves synaptic function
- Author
-
Yukiko Yoshida, Fei Chen, Bing Gong, Yong Pan, Giulio Maria Pasinetti, Isabel Arrieta-Cruz, and Vahram Haroutunian
- Subjects
Genetically modified mouse ,Aging ,Endoplasmic reticulum ,Long-term potentiation ,Cell Biology ,Biology ,F-box protein ,Cell biology ,Ubiquitin ligase ,Biochemistry ,mental disorders ,biology.protein ,Amyloid precursor protein ,Amyloid precursor protein secretase ,Intracellular - Abstract
BACE1 (β-secretase) plays a central role in the β-amyloidogenesis of Alzheimer's disease (AD). The ubiquitin-proteasome system, a major intracellular protein quality control system, has been implicated recently in BACE1 metabolism. We report that the SCF(Fbx2) -E3 ligase is involved in the binding and ubiquitination of BACE1 via its Trp 280 residue of F-box-associated domain. Physiologically, we found that Fbx2 was expressed in various intracellular organelles in brain neurons and that BACE1 is colocalized with Fbx2 and the amyloid precursor protein (APP), mainly at the early endosome and endoplasmic reticulum. The former are believed to be the major intracellular compartments where the APP is cleaved by BACE1 and β-amyloid is produced. Importantly, we found that overexpression of Fbx2 in the primary cortical and hippocampal neurons derived from Tg2576 transgenic mice significantly promoted BACE1 degradation and reduced β-amyloid production. In the search for specific endogenous modulators of Fbx2 expression, we found that PPARγ coactivator-1α (PGC-1α) was capable of promoting the degradation of BACE1 through a mechanism involving Fbx2 gene expression. Interestingly, we found that the expression of both Fbx2 and PGC-1α was significantly decreased in the brains of aging Tg2576 mice. Our in vivo studies using a mouse model of AD revealed that exogenous adenoviral Fbx2 expression in the brain significantly decreased BACE1 protein levels and activity, coincidentally reducing β-amyloid levels and rescuing synaptic deficits. Our study is the first to suggest that promoting Fbx2 in the brain may represent a novel strategy for the treatment of AD.
- Published
- 2010
43. A Pilot Study for the Effects of Donepezil Therapy on Cerebral and Optic Nerve Head Blood Flow, Visual Field Defect in Normal-Tension Glaucoma
- Author
-
Yasuharu Ogura, Tetsuya Sugiyama, Keita Utsunomiya, Tsunehiko Ikeda, and Yukiko Yoshida
- Subjects
Male ,medicine.medical_specialty ,Intraocular pressure ,genetic structures ,Optic Disk ,Vision Disorders ,Administration, Oral ,Glaucoma ,Pilot Projects ,Piperidines ,Donepezil Hydrochloride ,Ophthalmology ,Normal tension glaucoma ,Laser-Doppler Flowmetry ,medicine ,Humans ,Donepezil ,Pharmacology (medical) ,Low Tension Glaucoma ,Intraocular Pressure ,Aged ,Tomography, Emission-Computed, Single-Photon ,Pharmacology ,business.industry ,Brain ,Blood flow ,Iofetamine ,medicine.disease ,eye diseases ,Cerebral blood flow ,Regional Blood Flow ,Cerebrovascular Circulation ,Anesthesia ,Indans ,Optic nerve ,Female ,Cholinesterase Inhibitors ,sense organs ,Radiopharmaceuticals ,Visual Fields ,business ,Blood Flow Velocity ,medicine.drug - Abstract
To evaluate the effect of donepezil hydrochloride, an agent for the treatment of Alzheimer's disease (AD), on the cerebral and optic nerve head (ONH) blood flow, visual field defect in normal-tension glaucoma (NTG) patients with decreased cerebral blood flow (CBF) that demonstrates an AD-like perfusion pattern.The subjects were 5 NTG patients who exhibited AD-like decreased CBF upon (123)I-iodoamphetamine single photon emission computed tomography ((123)I-IMP SPECT). Donepezil hydrochloride (5 mg/day) was prescribed for each patient during a period of 12 months. Intraocular pressure (IOP), mean deviation (MD) of the Humphrey visual field, ONH blood flow determined by a laser speckle flowgraphy, and regional CBF (rCBF) determined by (123)I-IMP SPECT were measured before and every 6 months during the treatment.MD, ONH blood flow, and rCBF were improved significantly after 6 months of the treatment, although IOP did not change significantly. No deterioration of NTG morbidity was found in any of the measured parameters after 12 months of the treatment.Oral administration of donepezil hydrochloride in NTG patients might prevent deterioration of visual field defect, ONH blood flow, and rCBF in the temporal, parietal, and posterior lobes. This pilot study suggested the possibility that donepezil hydrochloride might ameliorate glaucomatous optic neuropathy in NTG.
- Published
- 2010
44. Cytosolic N-Glycans: Triggers for Ubiquitination Directing Proteasomal and Autophagic Degradation
- Author
-
Keiji Tanaka and Yukiko Yoshida
- Subjects
0301 basic medicine ,Proteasome Endopeptidase Complex ,Galectins ,Ubiquitin-Protein Ligases ,Cellular homeostasis ,General Biochemistry, Genetics and Molecular Biology ,Host-Parasite Interactions ,Tripartite Motif Proteins ,03 medical and health sciences ,Cytosol ,Polysaccharides ,Phagosomes ,Organelle ,Autophagy ,Humans ,Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase ,Secretory pathway ,030102 biochemistry & molecular biology ,biology ,Chemistry ,Macrophages ,Ubiquitination ,Nuclear Proteins ,Lectin ,Mycobacterium tuberculosis ,Cell biology ,DNA-Binding Proteins ,030104 developmental biology ,Secretory protein ,Proteasome ,biology.protein ,Protein Processing, Post-Translational ,Signal Transduction ,Transcription Factors - Abstract
Proteins on the cell surface and secreted proteins are modified with sugar chains that generate and modulate biological complexity and diversity. Sugar chains not only contribute physically to the conformation and solubility of proteins, but also exert various functions via sugar-binding proteins (lectins) that reside on the cell surface or in organelles of the secretory pathway. However, some glycosidases and lectins are found in the cytosol or nucleus. Recent studies of cytosolic sugar-related molecules have revealed that sugar chains on proteins in the cytosol act as signals of adverse cellular conditions. In this review, we summarize recent reports that cytosolic sugar chains can trigger ubiquitination, followed by proteasomal and autophagic degradation to maintain cellular homeostasis. In addition, we discuss the functions of sugar-binding proteins revealed to date, along with possibilities not yet explored.
- Published
- 2018
45. Prediction of Japanese children at risk for complications of childhood obesity : gender differences for intervention approaches
- Author
-
Yoichi Sato, Takuro Nakano, Hokuma Munakata, Mayumi Umeno, Masako Sei, Kozue Sakamoto, Ashraf A. Ewis, Chiemi Onishi, Yukiko Yoshida, and Yutaka Nakahori
- Subjects
Gerontology ,Male ,Risk ,medicine.medical_specialty ,life habits ,General Biochemistry, Genetics and Molecular Biology ,Childhood obesity ,Body Mass Index ,Intervention (counseling) ,medicine ,Humans ,Obesity ,Child ,intervention ,Sex Characteristics ,business.industry ,metabolic complications ,Public health ,Alanine Transaminase ,General Medicine ,Anthropometry ,medicine.disease ,Pedometer ,Female ,business ,Body mass index ,childhood obesity ,Sex characteristics - Abstract
Childhood obesity is one of the most serious public health problems in Japan, especially in Tokushima compared with other prefectures. This study was designed to clarify the life habits which predispose to development of obesity and can be modified through an appropriate intervention program to combat childhood obesity and its lifestyle-related diseases. A total of 216 school children from Itano Town, a municipality of Tokushima Prefecture, Japan, who are attending the fourth grade (9-10 years) of elementary schools, participated in the study from 2004 to 2007. The study included child’s life habits questionnaire, investigating physical activity by recording the daily steps using a pedometer, anthropometric measurements, hematological examination and hemodynamometry in a cross-sectional survey during a two-month period from June to July every year. We conclude that there are considerable gender-related differences for developing obesity and other lifestyle-related diseases ; and all intervention strategies against obesity must consider such gender differences. For example, restriction of television watching hours must be intervened for controlling obesity in boys, however for girls, promotion of exercise practice or making more steps per day with adequate sleeping periods should be intervened as the proper approaches for preventing and controlling obesity and other lifestyle-related diseases.
- Published
- 2010
46. STUDY ON THE IMPROVEMENT OF AIR CONDITIONING SYSTEM USING MATHEMATICAL PRINCIPLE METHOD CONTROL TECHNOLOGY
- Author
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Yasumi Fujinuma and Yukiko Yoshida
- Subjects
Engineering ,Environmental Engineering ,business.industry ,Energy consumption ,Optimal control ,Automotive engineering ,Energy management system ,Climate change mitigation ,Real-time simulation ,Air conditioning ,Control system ,Electricity ,business ,Simulation - Abstract
Building and Energy Management System (BEMS) has being focused on climate change mitigation in the IPCC 4th report. In order to reduce CO2 emissions in commercial sector in Japan, the project driven by NIES, National Institute for Environmental Studies, has developed the next generation BEMS that is configured with the environment measurement, the calculation of building energy and mathematical planning, and equipment control. This study is performed as a part of air conditioning control system development by real time heat load and system simulation in the real building. Regional forecast weather data, number of persons staying in the room and electricity consumption are fed to this simulation for input data. Several sensors are set in this building for feeding input data of real time simulation. We studied the energy consumption of the office to simulate and forecast the energy saving and evaluated an office model using indoor air-conditioning control system. We tried to optimal control for the indoor office model.
- Published
- 2010
47. Crystallization and preliminary X-ray characterization of the Skp1–Fbg3 complex
- Author
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Keiji Tanaka, Tomomi Nakagawa, Yukiko Yoshida, Tsunehiro Mizushima, Fuminori Tokunaga, Kazuhiro Iwai, Takashi Yamane, Atsuo Suzuki, and Taichi Kumanomidou
- Subjects
Biophysics ,Biology ,Crystallography, X-Ray ,medicine.disease_cause ,Biochemistry ,F-box protein ,law.invention ,Structural Biology ,law ,Skp1 ,Genetics ,medicine ,Humans ,Molecule ,Crystallization ,Escherichia coli ,chemistry.chemical_classification ,SKP Cullin F-Box Protein Ligases ,F-Box Proteins ,X-ray ,Condensed Matter Physics ,Ubiquitin ligase ,Crystallography ,chemistry ,Crystallization Communications ,biology.protein ,Glycoprotein - Abstract
F-box proteins are the substrate-recognition components of Skp1-Cullin1-F-box protein-Rbx1 (SCF) ubiquitin ligase complexes. Fbs1, an F-box protein, binds specifically to proteins modified with high-mannose oligosaccharides. Fbg3, another F-box protein, has 51% sequence identity to Fbs1. Although the residues that are necessary for binding to oligosaccharides are conserved between Fbs1 and Fbg3, Fbg3 does not bind glycoproteins. Skp1 and Fbg3 were co-expressed in Escherichia coli and their complex was purified to homogeneity and crystallized. Microseeding combined with the sandwiched hanging-drop technique improved the quality of the resulting crystals. The plate-shaped crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 34.1, b = 76.6, c = 193.9 A and one molecule per asymmetric unit.
- Published
- 2009
48. Cell shape, cell–cell contact, cell–extracellular matrix contact and cell polarity are all required for the maximum induction of CYP2B1 and CYP2B2 gene expression by phenobarbital in adult rat cultured hepatocytes
- Author
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Yukiko Yoshida, Akino Kawamura, Hiroaki Oda, and Atsushi Kakinuma
- Subjects
Collagen Type IV ,Male ,Cytoskeleton organization ,Cell Count ,Cell Communication ,Biochemistry ,Collagen Type I ,Extracellular matrix ,Type IV collagen ,Cell–cell interaction ,Laminin ,Cell polarity ,Gene expression ,Animals ,Rats, Wistar ,Cell Shape ,Cells, Cultured ,Pharmacology ,biology ,Cell Polarity ,Extracellular Matrix ,Rats ,Cell biology ,Gene Expression Regulation ,Cell culture ,Phenobarbital ,Cytochrome P-450 CYP2B1 ,Steroid Hydroxylases ,Hepatocytes ,biology.protein ,Aryl Hydrocarbon Hydroxylases ,Gels - Abstract
The effect of cell shape, cell density, contact with extracellular matrix and cell polarity on the phenobarbital (PB)-induced gene expression of CYP2B1 and CYP2B2 (CYP2B1/2B2) in adult rat hepatocytes was investigated. High cell density enhanced the induction of CYP2B1/2B2 gene expression by PB. Hepatocytes cultured on EHS gel showed a spherical cell shape and highly enhanced the induction of CYP2B1/2B2 gene expression by PB. Although monolayer hepatocytes cultured on type I collagen (TIC) and type IV collagen exhibited poor induction of CYP2B1/2B2 gene expression by PB, monolayer cells on laminin showed substantial induction. The addition of soluble laminin to media did not show any effect on induction in monolayer hepatocytes cultured on TIC. Dishes coated with different concentrations of immovable laminin demonstrated complicated effects. Coating with higher concentrations of laminin resulted in greater induction of CYP2B1/2B2 gene expression by PB. On the other hand, when hepatocytes were cultured on dishes coated with lower concentrations of laminin, they became round and greater induction of CYP2B1/2B2 gene expression by PB was observed. Spherical hepatocytes cultured on low concentrations of TIC also showed highly enhanced induction of CYP2B1/2B2 gene expression by PB. EHS gel overlay to hepatocytes cultured on TIC and collagen sandwich configurations that are known to induce cell polarity enhanced the induction by PB. The induction of CYP2B1/2B2 gene expression needed cytoskeleton organization, such as actin filament, microtubule filament and intermediate filament. These results demonstrate that cell shape, cell density, contact with extracellular matrix and cell polarity all play critical roles in the induction of CYP2B1/2B2 gene expression by PB.
- Published
- 2008
49. F-Box Proteins That Contain Sugar-Binding Domains
- Author
-
Yukiko Yoshida
- Subjects
Proteasome Endopeptidase Complex ,GTPase-activating protein ,Ubiquitin-Protein Ligases ,Molecular Sequence Data ,Carbohydrates ,Cell Cycle Proteins ,Nerve Tissue Proteins ,Applied Microbiology and Biotechnology ,Biochemistry ,F-box protein ,Analytical Chemistry ,Protein structure ,Ubiquitin ,Humans ,Amino Acid Sequence ,Molecular Biology ,Peptide sequence ,Glycoproteins ,chemistry.chemical_classification ,biology ,F-Box Proteins ,Organic Chemistry ,General Medicine ,Protein Structure, Tertiary ,Ubiquitin ligase ,Cell biology ,Membrane protein ,chemistry ,biology.protein ,Carbohydrate Metabolism ,Glycoprotein ,Biotechnology - Abstract
F-box proteins are the substrate-recognition subunits of the SCF (Skp1-Cul1-F-box protein) complex, which is the largest known class of E3 ubiquitin ligases. They play important roles in ubiquitin-dependent proteolysis in eukaryotes. The human genome contains about 70 genes for F-box proteins, and at least five homologous F-box proteins containing a conserved motif in their C-termini are thought to recognize sugar chain of N-linked glycoproteins. Among theses, Fbs1 and Fbs2 are perhaps involved in the endoplasmic reticulum-associated degradation pathway. In this review, I focus on the in vivo function of Fbs1 and homologous proteins, novel intracellular oligosaccharide recognition molecules involved in the quality control system.
- Published
- 2007
50. Structural basis for the selection of glycosylated substrates by SCF Fbs1 ubiquitin ligase
- Author
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Takashi Yamane, Tsunehiro Mizushima, Atsuo Suzuki, Yukiko Yoshida, Taichi Kumanomidou, Keiji Tanaka, and Yuko Hasegawa
- Subjects
Models, Molecular ,Glycosylation ,Protein family ,Molecular Sequence Data ,Static Electricity ,Oligosaccharides ,Ubiquitin-conjugating enzyme ,Crystallography, X-Ray ,Endoplasmic Reticulum ,Protein Structure, Secondary ,Substrate Specificity ,Ribonucleases ,Protein structure ,Ubiquitin ,SKP Cullin F-Box Protein Ligases ,Carbohydrate Conformation ,Escherichia coli ,Animals ,Humans ,Amino Acid Sequence ,Glycoproteins ,chemistry.chemical_classification ,DNA ligase ,Binding Sites ,Multidisciplinary ,biology ,Hydrogen Bonding ,Biological Sciences ,Protein Structure, Tertiary ,Ubiquitin ligase ,Carbohydrate Sequence ,Models, Chemical ,chemistry ,Biochemistry ,Ubiquitin ligase complex ,biology.protein ,Mannose ,Protein Binding - Abstract
The ubiquitin ligase complex SCF Fbs1 , which contributes to the ubiquitination of glycoproteins, is involved in the endoplasmic reticulum-associated degradation pathway. In SCF ubiquitin ligases, a diverse array of F-box proteins confers substrate specificity. Fbs1/Fbx2, a member of the F-box protein family, recognizes high-mannose oligosaccharides. To elucidate the structural basis of SCF Fbs1 function, we determined the crystal structures of the Skp1–Fbs1 complex and the sugar-binding domain (SBD) of the Fbs1–glycoprotein complex. The mechanistic model indicated by the structures appears to be well conserved among the SCF ubiquitin ligases. The structure of the SBD–glycoprotein complex indicates that the SBD primarily recognizes Man 3 GlcNAc 2 , thereby explaining the broad activity of the enzyme against various glycoproteins. Comparison of two crystal structures of the Skp1–Fbs1 complex revealed the relative motion of a linker segment between the F-box and the SBD domains, which might underlie the ability of the complex to recognize different acceptor lysine residues for ubiquitination.
- Published
- 2007
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