21 results on '"Sakurai, Keisuke"'
Search Results
2. Hyper BOLD Activation in Dorsal Raphe Nucleus of APP/PS1 Alzheimer's Disease Mouse during Reward-Oriented Drinking Test under Thirsty Conditions.
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Sakurai, Keisuke, Shintani, Teppei, Jomura, Naohiro, Matsuda, Takeshi, Sumiyoshi, Akira, and Hisatsune, Tatsuhiro
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ALZHEIMER'S disease , *HYPERPHAGIA , *BRAIN physiology , *PHENOTYPES , *VISUALIZATION - Abstract
Alzheimer's disease (AD), a neurodegenerative disease, causes behavioural abnormalities such as disinhibition, impulsivity, and hyperphagia. Preclinical studies using AD model mice have investigated these phenotypes by measuring brain activity in awake, behaving mice. In this study, we monitored the behavioural alterations of impulsivity and hyperphagia in middle-aged AD model mice. As a behavioural readout, we trained the mice to accept a water-reward under thirsty conditions. To analyse brain activity, we developed a measure for licking behaviour combined with visualisation of whole brain activity using awake fMRI. In a water-reward learning task, the AD model mice showed significant hyperactivity of the dorsal raphe nucleus in thirsty conditions. In summary, we successfully visualised altered brain activity in AD model mice during reward-oriented behaviour for the first time using awake fMRI. This may help in understanding the causes of behavioural alterations in AD patients. [ABSTRACT FROM AUTHOR]
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- 2020
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- View/download PDF
3. Total Synthesis of (−)‐Enigmazole A.
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Sakurai, Keisuke, Sasaki, Makoto, and Fuwa, Haruhiko
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MACROLIDE antibiotics synthesis , *MOIETIES (Chemistry) , *MACROCYCLIC compounds , *ALKENES - Abstract
Abstract: Total synthesis of (−)‐enigmazole A, a marine macrolide natural product with cytotoxic activity, has been accomplished. The tetrahydropyran moiety was constructed by means of a domino olefin cross‐metathesis/intramolecular oxa‐Michael addition of a δ‐hydroxy olefin. After coupling of advanced intermediates, the macrocycle was formed through gold‐catalyzed rearrangement of a propargylic benzoate, followed by ring‐closing metathesis of the resultant α,β‐unsaturated ketone. [ABSTRACT FROM AUTHOR]
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- 2018
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4. Total Synthesis of (-)-Enigmazole A.
- Author
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Sakurai, Keisuke, Sasaki, Makoto, and Fuwa, Haruhiko
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CHEMICAL synthesis , *NATURAL products , *NUCLEAR magnetic resonance spectroscopy , *ANTINEOPLASTIC agents , *METATHESIS reactions - Abstract
Total synthesis of (−)‐enigmazole A, a marine macrolide natural product with cytotoxic activity, has been accomplished. The tetrahydropyran moiety was constructed by means of a domino olefin cross‐metathesis/intramolecular oxa‐Michael addition of a δ‐hydroxy olefin. After coupling of advanced intermediates, the macrocycle was formed through gold‐catalyzed rearrangement of a propargylic benzoate, followed by ring‐closing metathesis of the resultant α,β‐unsaturated ketone. [ABSTRACT FROM AUTHOR]
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- 2018
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- View/download PDF
5. Structural basis for the inhibition of bacterial multidrug exporters.
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Nakashima, Ryosuke, Sakurai, Keisuke, Yamasaki, Seiji, Hayashi, Katsuhiko, Nagata, Chikahiro, Hoshino, Kazuki, Onodera, Yoshikuni, Nishino, Kunihiko, and Yamaguchi, Akihito
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MULTIDRUG transporters , *CRYSTAL structure , *GRAM-negative bacteria , *PSEUDOMONAS aeruginosa , *PYRIMIDINE derivatives , *MULTIDRUG resistance , *PROTEINS , *PHENYLALANINE - Abstract
The multidrug efflux transporter AcrB and its homologues are important in the multidrug resistance of Gram-negative pathogens. However, despite efforts to develop efflux inhibitors, clinically useful inhibitors are not available at present. Pyridopyrimidine derivatives are AcrB- and MexB-specific inhibitors that do not inhibit MexY; MexB and MexY are principal multidrug exporters in Pseudomonas aeruginosa. We have previously determined the crystal structure of AcrB in the absence and presence of antibiotics. Drugs were shown to be exported by a functionally rotating mechanism through tandem proximal and distal multisite drug-binding pockets. Here we describe the first inhibitor-bound structures of AcrB and MexB, in which these proteins are bound by a pyridopyrimidine derivative. The pyridopyrimidine derivative binds tightly to a narrow pit composed of a phenylalanine cluster located in the distal pocket and sterically hinders the functional rotation. This pit is a hydrophobic trap that branches off from the substrate-translocation channel. Phe 178 is located at the edge of this trap in AcrB and MexB and contributes to the tight binding of the inhibitor molecule through a π-π interaction with the pyridopyrimidine ring. The voluminous side chain of Trp 177 located at the corresponding position in MexY prevents inhibitor binding. The structure of the hydrophobic trap described in this study will contribute to the development of universal inhibitors of MexB and MexY in P. aeruginosa. [ABSTRACT FROM AUTHOR]
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- 2013
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6. Cone Phosphodiesterase-6α' Restores Rod Function and Confers Distinct Physiological Properties in the Rod Phosphodiesterase-6β-Deficient rd1O Mouse.
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Wen-Tao Deng, Sakurai, Keisuke, Kolandaivelu, Saravanan, Kolesnikov, Alexander V., Dinculescu, Astra, Jie Li, Ping Zhu, Xuan Liu, Jijing Pang, Chiodo, Vince A., Boye, Sanford L., Bo Chang, Ramamurthy, Visvanathan, Kefalov, Vladimir J., and Hauswirth, William W.
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PHOSPHODIESTERASES , *GENETIC transduction , *RETINAL rod photoreceptor cells , *PHOTORECEPTORS , *ADENO-associated virus - Abstract
Phosphodiesterase-6 (PDE6) is the key effector enzyme of the vertebrate phototransduction pathway in rods and cones. Rod PDE6 catalytic core is composed of two distinct subunits, PDE6α and PDE6β, whereas two identical PDE6α' subunits form the cone PDE6 catalytic core. It is not known whether this difference in PDE6 catalytic subunit identity contributes to the functional differences between rods and cones. To address this question, we expressed cone PDE6α' in the photoreceptor cells of the retinal degeneration 10 (rdlO) mouse that carries a mutation in rod PDEβ subunit. We show that adeno-associated virus-mediated subretinal delivery of PDE6α' rescues rod electroretinogram responses and preserves retinal structure, indicating that cone PDE6α' can couple effectively to the rod phototransduction pathway. We also show that restoration of light sensitivity in rdlO rods is attributable to assembly of PDE6α' with rod PDE6γ. Single-cell recordings revealed that, surprisingly, rods expressing cone PDE6α' are twofold more sensitive to light than wild-type rods, most likely because of the slower shutoff of their light responses. Unlike in wild-type rods, the response kinetics in PDE6α' -treated rdl 0 rods accelerated with increasing flash intensity, indicating a possible direct feedback modulation of cone PDE6α' activity. Together, these results demonstrate that cone PDE6α' can functionally substitute for rod PDEαβ in vivo, conferring treated rods with distinct physiological properties. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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7. Structures of the multidrug exporter AcrB reveal a proximal multisite drug-binding pocket.
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Nakashima, Ryosuke, Sakurai, Keisuke, Yamasaki, Seiji, Nishino, Kunihiko, and Yamaguchi, Akihito
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GRAM-negative bacteria , *DRUG tolerance , *RIFAMPIN , *ERYTHROMYCIN , *DOXORUBICIN , *MONOMERS - Abstract
AcrB and its homologues are the principal multidrug transporters in Gram-negative bacteria and are important in antibiotic drug tolerance. AcrB is a homotrimer that acts as a tripartite complex with the outer membrane channel TolC and the membrane fusion protein AcrA. Minocycline and doxorubicin have been shown to bind to the phenylalanine cluster region of the binding monomer. Here we report the crystal structures of AcrB bound to the high-molecular-mass drugs rifampicin and erythromycin. These drugs bind to the access monomer, and the binding sites are located in the proximal multisite binding pocket, which is separated from the phenylalanine cluster region (distal pocket) by the Phe-617 loop. Our structures indicate that there are two discrete multisite binding pockets along the intramolecular channel. High-molecular-mass drugs first bind to the proximal pocket in the access state and are then forced into the distal pocket in the binding state by a peristaltic mechanism involving subdomain movements that include a shift of the Phe-617 loop. By contrast, low-molecular-mass drugs, such as minocycline and doxorubicin, travel through the proximal pocket without specific binding and immediately bind to the distal pocket. The presence of two discrete, high-volume multisite binding pockets contributes to the remarkably broad substrate recognition of AcrB. [ABSTRACT FROM AUTHOR]
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- 2011
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8. Role of Guanylyl Cyclase Modulation in Mouse Cone Phototransduction.
- Author
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Sakurai, Keisuke, Jeannie Chen, and Kefalov, Vladimir J.
- Abstract
A negative phototransduction feedback in rods and cones is critical for the timely termination of their light responses and for extending their function to a wide range of light intensities. The calcium feedback mechanisms that modulate phototransduction in rods have been studied extensively. However, the corresponding modulation mechanisms that enable cones to terminate rapidly their light responses and to adapt in bright light, properties critical for our daytime vision, are still not understood. In cones, calcium feedback to guanylyl cyclase is potentially a key step in phototransduction modulation. The guanylyl cyclase activity is modulated by the calcium-binding guanylyl cyclase activating proteins (GCAP1 and GCAP2). Here, we used single-cell andtransretinal recordings from mouseto determine how GCAPs modulate dark-adapted responses as well as light adaptation in mammalian cones. Deletion of GCAPs increasedthreefoldthe amplitude and dramatically prolonged the light responses in dark-adapted mouse cones. It also reduced the operating range of mouse cones in background illumination and severely impaired their light adaptation. Thus, GCAPs exert powerful modulation on the mammalian cone phototransduction cascade and play an important role in setting the functional properties of cones in darkness and during light adaptation. Surprisingly, despite their better adaptation capacity and wider calcium dynamic range, mammalian cones were modulated by GCAPsto a lesser extentthan mammalian rods.We concludethat a disparity inthe strength of GCAP modulation cannot explain the differences in the dark-adapted properties or in the operating ranges of mammalian rods and cones. [ABSTRACT FROM AUTHOR]
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- 2011
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9. Deletion of GRK1 Causes Retina Degeneration through a Transducin-Independent Mechanism.
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Fan, Jie, Sakurai, Keisuke, Ching-Kang Chen, Rohrer, Baerbel, Wu, Bill X., King-Wai Yau, Kefalov, Vladimir, and Crouch, Rosalie K.
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RETINAL degeneration , *RETINOIDS , *PHOSPHORYLATION , *LABORATORY mice , *OPSINS - Abstract
Rpe65-/- mice are unable to produce 11-cis-retinal, the chromophore of visual pigments. Consequently, the pigment is present as the apoprotein opsin with a minute level of pigment containing 9-cis-retinal as chromophore. Notably, a 10-20% fraction of this opsin is mono-phosphorylated independently of light conditions. To determine the role of rhodopsin kinase (GRK1) in phosphorylating this opsin and to test whether eliminating this phosphorylation would accelerate photoreceptor degeneration, we generated the Rpe65-/-Grk1-/- mouse. The retinae of Rpe65-/-Grk1-/- mice had negligible opsin phosphorylation, extensive degeneration with decreased opsin levels, and diminished light-evoked rod responses relative to Rpe65-/- mice. These data show that opsin phosphorylation in the Rpe65-/-mouse is due to the action of GRK1 and is neuroprotective. However, despite the higher activity of unphosphorylated opsin, the severe loss of opsin in the rapidly degenerating Rpe65-/-Grk1-/- mice resulted in lower overall opsin activity and in higher rod sensitivity compared with Rpe65-/- mice. In Rpe65-/-Grk1-/-Gnat1-/- mice where transduction activation was blocked, degeneration was only partially prevented. Therefore, increased opsin activity in the absence of phosphorylation was not the only mechanism for the accelerated retinal degeneration. Finally, the deletion of GRK1 triggered retinal degeneration in Grk1-/- mice after 1 month, even in the absence of apo-opsin. This degeneration was independent of light conditions and occurred even in the absence of transducin in Grk1-/-Gnat1-/- mice. Taken together, our results demonstrate a light-independent mechanism for retinal degeneration in the absence of GRK1, suggesting a second, not previously recognized role for that kinase. [ABSTRACT FROM AUTHOR]
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- 2010
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10. Substrate binding induces structural changes in cytochrome P450cam.
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Sakurai, Keisuke, Shimada, Hideo, Hayashi, Takashi, and Tsukihara, Tomitake
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CYTOCHROME P-450 , *HYDROGEN bonding , *X-ray crystallography , *OXIDATION-reduction reaction , *BIOCHEMICAL substrates - Abstract
The binding of (+)-camphor to cytochrome P450cam (P450cam) expels a cluster of waters at the active site, raising the redox potential of the haem to an extent that allows reduction by the electron-transfer system. This binding was reported to involve no significant structural changes in the protein. Here, two ferric P450cam structures partially complexed with (+)-camphor were determined by X-ray crystallography at 1.30-1.35 Å resolution, revealing the structures of the substrate-free and substrate-bound forms. (+)-Camphor binding induces rotation of Thr101 to form a hydrogen bond that acts as a hydrogen donor to a peripheral haem propionate. This bonding contributes to the redox-potential change. [ABSTRACT FROM AUTHOR]
- Published
- 2009
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11. Physiological Properties of Rod Photoreceptor Cells in Green-sensitive Cone Pigment Knock-in Mice.
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Sakurai, Keisuke, Onishi, Akishi, Imai, Hiroo, Chisaka, Osamu, Ueda, Yoshiki, Usukura, Jiro, Nakatani, Kei, and Shichida, Yoshinori
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PHOTORECEPTORS , *LABORATORY mice , *VISUAL pigments , *RETINA , *RHODOPSIN , *MICROBIAL genetics - Abstract
Rod and cone photoreceptor cells that are responsible for scotopic and photopic vision, respectively, exhibit photo-responses different from each other and contain similar phototransduction proteins with distinctive molecular properties. To investigate the contribution of the different molecular properties of visual pigments to the responses of the photoreceptor cells, we have generated knock-in mice in which rod visual pigment (rhodopsin) was replaced with mouse green-sensitive cone visual pigment (mouse green). The mouse green was successfully transported to the rod outer segments, though the expression of mouse green in homozygous retina was ∼11% of rhodopsin in wild-type retina. Single-cell recordings of wild-type and homozygous rods suggested that the flash sensitivity and the single-photon responses from mouse green were three to fourfold lower than those from rhodopsin after correction for the differences in cell volume and levels of several signal transduction proteins. Subsequent measurements using heterozygous rods expressing both mouse green and rhodopsin E122Q mutant, where these pigments in the same rod cells can be selectively irradiated due to their distinctive absorption maxima, clearly showed that the photo-response of mouse green was threefold lower than that of rhodopsin. Noise analysis indicated that the rate of thermal activations of mouse green was 1.7 X 10-7 s-1, about 860-fold higher than that of rhodopsin. The increase in thermal activation of mouse green relative to that of rhodopsin results in only 4% reduction of rod photosensitivity for bright lights, but would instead be expected to severely affect the visual threshold under dim-light conditions. Therefore, the abilities of rhodopsin to generate a large single photon response and to retain high thermal stability in darkness are factors that have been necessary for the evolution of scotopic vision. [ABSTRACT FROM AUTHOR]
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- 2007
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12. Relationship between elevated impulsivity and cognitive declines in elderly community-dwelling individuals.
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Sakurai, Keisuke, Li, Haowei, Inamura, Noriko, Masuoka, Nobutaka, and Hisatsune, Tatsuhiro
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DEMENTIA , *COGNITION in old age , *AGGRESSION (Psychology) , *AGITATION (Psychology) , *PSYCHOMETRICS - Abstract
Impulse control disorders are recognized as one of the behavioral and psychological symptoms of dementia (BPSD). Majority of studies on the treatment of BPSD related to impulsivity have rather focused on the aggression and agitation. In particular, it has not been investigated how cognitive declines are associated with impulsivity in community-dwelling elderly people. Here, we have measured the cognitive and memory functions and impulsivity of 212 elderly community-dwelling people using a psychometric test battery and analyzed the correlation between their level of impulsivity and cognitive functions by multiple regression analysis. We found an elevation of impulsivity, which was evaluated by the Barratt Impulsiveness Scale-11, closely related to decline of cognitive functions, which were evaluated by the Montreal Cognitive Assessment and the Mini-Mental State Examination, and Logical Memory function, which were evaluated by the Wechsler Memory Scale-Delayed Recall. Then we have divided them into groups based on the severity of cognitive decline and conducted an analysis of each group, the result of which showed that as this tendency was particularly noticeable in the suspected dementia group. Therefore, we have concluded that heightened impulsivity is negatively associated with cognitive and memory functions in community-dwelling elderly people. [ABSTRACT FROM AUTHOR]
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- 2020
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13. One-pot concise syntheses of benzimidazo[2,1-a]isoquinolines by a microwave-accelerated tandem process
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Okamoto, Noriko, Sakurai, Keisuke, Ishikura, Minoru, Takeda, Kei, and Yanada, Reiko
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ORGANIC synthesis , *BENZIMIDAZOLES , *ISOQUINOLINE , *MICROWAVES , *ALDEHYDES , *ALKYNES , *PHENYLENEDIAMINES , *RING formation (Chemistry) - Abstract
Abstract: Direct, efficient syntheses of the benzimidazo[2,1-a]isoquinoline ring system have been achieved with 2-bromoarylaldehydes, terminal alkynes, and 1,2-phenylenediamines by a microwave-accelerated tandem process in which a Sonogashira coupling, 5-endo cyclization, oxidative aromatization, and 6-endo cyclization can be performed in a single synthetic operation. [Copyright &y& Elsevier]
- Published
- 2009
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14. The Na+/Ca2+, K+ exchanger 2 modulates mammalian cone phototransduction.
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Sakurai, Keisuke, Vinberg, Frans, Wang, Tian, Chen, Jeannie, and Kefalov, Vladimir J.
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- 2016
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15. Evaluation of the Functional Role of the Heme-6-propionate Side Chain in Cytochrome P450cam.
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Harada, Katsuyoshi, Sakurai, Keisuke, Ikemura, Kenichiro, Ogura, Takashi, Hirota, Shun, Shimada, Hideo, and Hayashi, Takashi
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CHEMICAL research , *HEME , *PROPIONATES , *CYTOCHROME P-450 , *COORDINATION compounds , *PHYSICAL & theoretical chemistry - Abstract
The article presents a study which examines the functional role of the heme-6-propionate side chain in cytochrome P450cam. Results revealed that heme-6-propionate is significant in the fixation of the putidaredoxin (Pdx)-binding site by the Argon 112 residue and the stabilization of the iron (Fe)-sulphur (S) coordination to prevent the development of the inactive P420 species. Also, 6-propionate side chain interaction of the amino acid is essential to retain the enzyme activity of P450cam.
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- 2008
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16. An incremental online semi-supervised active learning algorithm based on self-organizing incremental neural network.
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Shen, Furao, Yu, Hui, Sakurai, Keisuke, and Hasegawa, Osamu
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COMPUTER algorithms , *SUPERVISED learning , *SELF-organizing systems , *ARTIFICIAL neural networks , *DATA structures , *EXPERIMENTS , *STATISTICAL sampling - Abstract
An incremental online semi-supervised active learning algorithm, which is based on a self-organizing incremental neural network (SOINN), is proposed. This paper describes improvement of the two-layer SOINN to a single-layer SOINN to represent the topological structure of input data and to separate the generated nodes into different groups and subclusters. We then actively label some teacher nodes and use such teacher nodes to label all unlabeled nodes. The proposed method can learn from both labeled and unlabeled samples. It can query the labels of some important samples rather than selecting the labeled samples randomly. It requires neither prior knowledge, such as the number of nodes, nor the number of classes. It can automatically learn the number of nodes and teacher vectors required for a current task. Moreover, it can realize online incremental learning. Experiments using artificial data and real-world data show that the proposed method performs effectively and efficiently. [ABSTRACT FROM AUTHOR]
- Published
- 2011
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17. Molecular Properties of Rhodopsin and Rod Function.
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Imai, Hiroo, Kefalov, Vladimir, Sakurai, Keisuke, Chisaka, Osamu, Uedam, Yoshiki, Onishi, Akishi, Morizumi, Takefumi, Yingbin Fu, Ichikawa, Kazuhisa, Nakatani, Kei, Hondam, Yoshihito, Chen, Jeannie, King-Wai Yau, and Shichida, Yoshinori
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CELLS , *RHODOPSIN , *PHOTONS , *CELLULAR signal transduction , *GENETIC mutation - Abstract
Signal transduction in rod cells begins with photon absorption by rhodopsin and leads to the generation of an electrical response. The response profile is determined by the molecular properties of the phototransduction components. To examine how the molecular properties of rhodopsin correlate with the rod-response profile, we have generated a knock-in mouse with rhodopsin replaced by its E122Q mutant, which exhibits properties different from those of wild-type (WT) rhodopsin. Knock-in mouse rods with E122Q rhodopsin exhibited a photo-sensitivity about 70% of WT. Correspondingly, their single-photon response had an amplitude about 80% of WT, and a rate of decline from peak about 1.3 times of WT. The overall 30% lower photosensitivity of mutant rods can be explained by a lower pigment photosensitivity (0.9) and the smaller single-photon response (0.8). The slower decline of the response, however, did not correlate with the 10-fold shorter lifetime of the meta-!! state of E122Q rhodopsin. This shorter lifetime became evident in the recovery phase of rod cells only when arrestin was absent. Simulation analysis of the photoresponse profile indicated that the slower decline and the smaller amplitude of the single-photon response can both be explained by the shift in the meta-I/meta-II equilibrium of E122Q rhodopsin toward meta-I. The difference in meta-Ill lifetime between WT and E122Q mutant became obvious in the recovery phase of the dark current after moderate photobleaching of rod cells. Thus, the present study clearly reveals how the molecular properties of rhodopsin affect the amplitude, shape, and kinetics of the rod response. [ABSTRACT FROM AUTHOR]
- Published
- 2007
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18. A Role of the Heme-7-Propionate Side Chain in Cytochrome P450cam as a Gate for Regulating the Access of Water Molecules to the Substrate-Binding Site.
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Hayashi, Takashi, Harada, Katsuyoshi, Sakurai, Keisuke, Shimada, Hideo, and Hirota, Shun
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CYTOCHROME P-450 , *SUBSTRATES (Materials science) , *WATER , *MOLECULES , *MONOOXYGENASES , *PROPIONATES , *CAMPHOR - Abstract
The article focuses on the role of heme-7-propionate side chain for controlling the access of water molecules in Cytochrome P450cam. Cytochrome P450cam is a monooxygenase that uses thiolate binding heme b as a prosthetic group. It proposes the use of 7-propionate side chain that can be used to repel water molecules to increase high d-camphor affinity and to regulate water expulsion system at the starting point of the catalytic cycle of P450cam.
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- 2009
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19. Crystal structure of the multidrug resistance regulator RamR complexed with bile acids.
- Author
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Yamasaki, Suguru, Nakashima, Ryosuke, Sakurai, Keisuke, Baucheron, Sylvie, Giraud, Etienne, Doublet, Benoît, Cloeckaert, Axel, and Nishino, Kunihiko
- Abstract
During infection, Salmonella senses and responds to harsh environments within the host. Persistence in a bile-rich environment is important for Salmonella to infect the small intestine or gallbladder and the multidrug efflux system AcrAB-TolC is required for bile resistance. The genes encoding this system are mainly regulated by the ramRA locus, which is composed of the divergently transcribed ramA and ramR genes. The acrAB and tolC genes are transcriptionally activated by RamA, whose encoding gene is itself transcriptionally repressed by RamR. RamR recognizes multiple drugs; however, the identity of the environmental signals to which it responds is unclear. Here, we describe the crystal structures of RamR in complexes with bile components, including cholic acid and chenodeoxycholic acid, determined at resolutions of 2.0 and 1.8 Å, respectively. Both cholic and chenodeoxycholic acids form four hydrogen bonds with Tyr59, Thr85, Ser137 and Asp152 of RamR, instead of π-π interactions with Phe155, a residue that is important for the recognition of multiple compounds including berberine, crystal violet, dequalinium, ethidium bromide and rhodamine 6 G. Binding of these compounds to RamR reduces its DNA-binding affinity, resulting in the increased transcription of ramA and acrAB-tolC. Our results reveal that Salmonella senses bile acid components through RamR and then upregulates the expression of RamA, which can lead to induction of acrAB-tolC expression with resulting tolerance to bile-rich environments. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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20. GABA-Induced GnRH Release Triggers Chordate Metamorphosis.
- Author
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Hozumi, Akiko, Matsunobu, Shohei, Mita, Kaoru, Treen, Nicholas, Sugihara, Takaho, Horie, Takeo, Sakuma, Tetsushi, Yamamoto, Takashi, Shiraishi, Akira, Hamada, Mayuko, Satoh, Noriyuki, Sakurai, Keisuke, Satake, Honoo, and Sasakura, Yasunori
- Subjects
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METAMORPHOSIS , *GONADOTROPIN , *GABA , *ECOLOGICAL niche , *GABA receptors , *FUNCTIONAL analysis - Abstract
Metamorphosis, a widespread life history strategy in metazoans, allows dispersal and use of different ecological niches through a dramatic body change from a larval stage [ 1 , 2 ]. Despite its conservation and importance, the molecular mechanisms underlying its initiation and progression have been characterized in only a few animal models. In this study, through pharmacological and gene functional analyses, we identified neurotransmitters responsible for metamorphosis of the ascidian Ciona. Ciona metamorphosis converts swimming tadpole larvae into vase-like, sessile adults. Here, we show that the neurotransmitter GABA is a key regulator of metamorphosis. We found that gonadotropin-releasing hormone (GnRH) is a downstream neuropeptide of GABA. Although GABA is generally thought of as an inhibitory neurotransmitter, we found that it positively regulates secretion of GnRH through the metabotropic GABA receptor during Ciona metamorphosis. GnRH is necessary for reproductive maturation in vertebrates, and GABA is an important excitatory regulator of GnRH in the hypothalamus during puberty [ 3 , 4 ]. Our findings reveal another role of the GABA-GnRH axis in the regulation of post-embryonic development in chordates. • The neurotransmitter GABA is a key regulator of Ciona metamorphosis • Gonadotropin-releasing hormone (GnRH) is the downstream neuropeptide of GABA • GABA positively regulates secretion of GnRH through the metabotropic GABA receptor Metamorphosis is a dramatic body change in metazoans that allows the use of different ecological niches. Hozumi et al. show that GABA is a key regulator of metamorphosis in the ascidian Ciona. During this process, GABA, generally thought of as an inhibitory neurotransmitter, positively regulates secretion of the neuropeptide GnRH. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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21. Acid-Tolerant Monomeric GFP from Olindias formosa.
- Author
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Shinoda, Hajime, Ma, Yuanqing, Nakashima, Ryosuke, Sakurai, Keisuke, Matsuda, Tomoki, and Nagai, Takeharu
- Subjects
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FLUORESCENT proteins , *PHOTOLUMINESCENCE , *X-ray diffraction , *IMMUNOFLUORESCENCE , *DYES & dyeing - Abstract
Summary The fluorescent protein (FP) color palette has greatly contributed to the visualization of molecular and cellular processes. However, most FPs lose fluorescence at a pH lower than their neutral p K a (∼6), and this has hampered their application in acidic organelles (pH ∼4.5–6.0). Currently, several cyan- and red-colored acid-tolerant FPs are available; however, there are few reports of acid-tolerant green FPs (GFPs) that are practically applicable to bioimaging. Here, we developed the acid-tolerant monomeric GFP “Gamillus” from the jellyfish Olindias formosa , with excellent brightness, maturation speed, and photostability. Results from X-ray crystallography and point mutagenesis suggest that across a broad pH range the acid tolerance is attributed to stabilization of deprotonation in the chromophore phenyl ring by forming a unique trans configuration. We demonstrate that Gamillus can serve as a molecular tag suitable for imaging in acidic organelles through autophagy-mediated molecular tracking to lysosomes. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
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