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OALib Journal期刊

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Biotinylated-sortase self-cleavage purification (BISOP) method for cell-free produced proteins
Satoko Matsunaga, Kazuhiro Matsuoka, Kouhei Shimizu, Yaeta Endo, Tatsuya Sawasaki
BMC Biotechnology , 2010, DOI: 10.1186/1472-6750-10-42
Abstract: The histidine tag-based self-cleavage method for purifying proteins produced by the wheat cell-free protein synthesis system showed high background, low recovery, and unexpected cleavage between the N-terminally fused sortase and target protein during the protein synthesis. Addition of calcium chelator BAPTA to the cell-free reaction inhibited the cleavage. In order to adapt the sortase-based purification method to the cell-free system, we next used biotin as the affinity tag. The biotinylated sortase self-cleavage purification (BISOP) method provided tag-free, highly purified proteins due to improved recovery of proteins from the resin. The N-terminal sequence analysis of the GFP produced by the BISOP method revealed that the cleavage indeed occurred at the right cleavage site. Using this method, we also successfully purified the E2 heterocomplex of USE2N and USE2v1. The c-terminal src kinase (CSK) obtained by the BISOP method showed high activity in phosphorylating the Src protein. Furthermore, we demonstrated that this method is suitable for automatically synthesizing and purifying proteins using robots.We demonstrated that the newly developed BISOP method is very useful for obtaining high quality, tag-free recombinant proteins, produced using the cell-free system, for biochemical and structural analyses.Technology used for purifying a recombinant protein has a significant impact on its biochemical function, structural properties, and other aspects, such as generating an antibody against the protein. Currently available, established purification methods generally attach an affinity tag to the N-terminus or C-terminus end of the target protein, and then recover the target protein by affinity chromatography [1]. Purification tags used today are classified into peptide-tags and protein-tags based on their nature and form. His-tag, a typical and globally the most used peptide-tag, is highly effective in purifying the tagged protein by using immobilized metal affinity
Association of APOBEC3G genotypes and CD4 decline in Thai and Cambodian HIV-infected children with moderate immune deficiency
Bunupuradah Torsak,Imahashi Mayumi,Iampornsin Thatri,Matsuoka Kazuhiro
AIDS Research and Therapy , 2012, DOI: 10.1186/1742-6405-9-34
Abstract: Introduction Human APOBEC3G is a host defense factor that potently inhibits HIV replication. We hypothesize that HIV-infected children with a genetic variant of APOBEC3G will have a more rapid disease progression. Methods Antiretroviral therapy (ART)-na ve children, aged 1–12 years old with CD4 15-24% and without severe HIV-related symptoms were enrolled. The children had CD4% and absolute CD4 counts every 12 weeks and HIV-RNA every 24 weeks until 144 weeks. ART was started when CD4% declined to < 15% or AIDS-related events developed. APOBEC3G genetic variants were performed by PCR-based restriction fragment length polymorphism techniques from peripheral blood mononuclear cells. Random-effect linear regression analysis was performed to correlate APOBEC3G genotypes and disease progression. Results 147 children, 35% male, with a median (IQR) age of 6.5 (4.3-8.8) years were enrolled. CDC N:A:B were 1:63:36%. Median baseline values were 20% for CD4% 605 cells/mm3 for CD4 count and 4.7 log10copies/mL for HIV-RNA. The frequencies of APOBEC3G genotypes AA (186H/H), AG (186H/R), GG (186R/R) were 86%, 12%, and 2% respectively. The APOBEC3G genotype GG was associated with a significant decline in CD4% -5.1% ( 8.9 to 1.2%), p<0.001, and CD4 counts 226 ( 415 to 34) cells/mm3, p<0.001 by random-effect liner regression analysis. No significant associations of APOBEC3G genotypes with HIV-RNA changes overtime (p=0.16) or progression to CDC B and C (p=0.49) were observed. Conclusions APOBEC3G genotype GG was significantly associated with a more rapid decline in CD4. APOBEC3G’s antiviral effects on HIV disease progression in children should be further explored.
Fabrication of Thermo-Responsive Molecular Layers from Self-Assembling Elastin-Like Oligopeptides Containing Cell-Binding Domain for Tissue Engineering
Tomoyuki Koga,Kazuhiro Nakamoto,Koji Odawara,Tomoo Matsuoka,Nobuyuki Higashi
Polymers , 2015, DOI: 10.3390/polym7010134
Abstract: Novel thermo-responsive elastin-like oligopeptides containing cell-binding epitope (Arg-Gly-Asp-Ser sequence); arginine-glycine-aspartic acid-serine (RGDS)-elastin-like peptides (ELP) and RGDS- deg-ELP; were newly prepared as building blocks of self-assembled molecular layer for artificial extra cellular matrix. A detailed analysis of the conformation of the oligo(ELP)s in water and their self-assembling behavior onto hydrophobic surfaces were performed by using circular dichroism, Fourier transform infrared spectroscopy (FTIR), atomic force microscopy and water contact angle measurements. The experimental results revealed that both oligo(ELP)s self-assembled onto hydrophobic surfaces and formed molecular layers based on their thermo-responsive conformational change from hydrous random coil to dehydrated β-turn structure. Effective cell adhesion and spreading behaviors were observed on these self-assembled oligo(ELP) layers. In addition, attached cells were found to be recovered successfully as a cell-sheet by temperature-induced disassembly of oligo(ELP) layer. This achievement provides an important insight to construct novel oligopeptide-based nano-surfaces for the design of smart artificial extra-cellular matrix.
Spontaneous Rupture of the Omental Artery Treated by Transcatheter Arterial Embolization
Masamichi Takahashi,Yujiro Matsuoka,Tsuyoshi Yasutake,Hiroyuki Abe,Kazuhiro Sugiyama,Kazuyuki Oyama
Case Reports in Radiology , 2012, DOI: 10.1155/2012/273027
Abstract: Intra-abdominal hemorrhage caused by omental artery rupture is a rare condition. There are few reports on the treatment of omental artery rupture with only transcatheter arterial embolization (TAE). A 27-year-old man presented to our emergency room with upper abdominal pain that suddenly occurred during sleep. Abdominal computed tomography (CT) revealed fluid collection in the peritoneal cavity and a left subphrenic hematoma with extravasation. Celiac angiography revealed extravasation from the omental artery, which arose from the proximal left gastroepiploic artery. A microcatheter was advanced into the left gastroepiploic artery and around the culprit artery bifurcation, which was embolized by inserting coils. The postoperative course was uneventful without worsening of anemia or abdominal symptoms. The patient was discharged after the absence of extravasation was confirmed by contrast-enhanced CT. Although surgical therapy has often been performed for omental bleeding, TAE, which is less invasive and has the advantage of simultaneous diagnosis and treatment, should be attempted as the first-choice therapy. 1. Introduction Rupture of the omental artery is a relatively rare condition that can lead to life-threatening intra-abdominal hemorrhage [1]. Causes of omental artery rupture include penetrating or blunt abdominal trauma, neoplasia [2], arterial aneurysm rupture [3], and complications of anticoagulant therapy [4]. However, it rarely develops without any specific triggers [5, 6]. We found only four reports on transcatheter arterial embolization (TAE) for rupture of the omental artery [7–10]. Along with a review of these reports, we herein present a case of omental artery rupture treated by TAE in a young patient who had no history of a related disease or trauma. 2. Case Presentation A 27-year-old man presented to our emergency unit after having been woken from sleep at midnight because of severe abdominal pain. His medical and familial histories were unremarkable. His vital signs and laboratory evaluation results, including the hemoglobin level (15.2？g/dL), showed no abnormalities. He was suspected to have gastritis and was discharged with an H2-blocker. However, the abdominal pain did not improve. It worsened when stretching his back and breathing. He returned to our hospital because of temporary loss of consciousness when rising from his bed the next morning. Contrast-enhanced CT revealed high-density ascites retention that was thought to contain blood. There was a left subphrenic hematoma compressing the stomach and spleen with extravasation
Suzaku Studies of Luminosity-Dependent Changes in the Low-Mass X-ray Binary Aquila X-1
Soki Sakurai,Shunsuke Torii,Hirofumi Noda,Zhongli Zhang,Ko Ono,Kazuhiro Nakazawa,Kazuo Makishima,Hiromitsu Takahashi,Shinya Yamada,Masaru Matsuoka
Physics , 2013, DOI: 10.1093/pasj/pst010
Abstract: The neutron-star Low-Mass X-ray Binary Aquila X-1 was observed by Suzaku for seven times, from 2007 September 28 to October 30. The observations successfully traced an outburst decay in which the source luminosity decreased almost monotonically from $\sim 10^{37}$ erg s$^{-1}$ to $\sim 10^{34}$ erg s$^{-1}$, by $\sim 3$ orders of magnitude. To investigate luminosity-dependent changes in the accretion geometry, five of the seven data sets with a typical exposure of $\sim 18$ ks each were analyzed; the other two were utilized in a previous work \citep{Sakurai2012}. The source was detected up to 100 keV in the 2nd to the 4th observations, to 40 keV in the 5th, and to 10 keV on the last two occasions. All spectra were reproduced successfully by Comptonized blackbody model with relatively high ($\gtrsim 2.0$) optical depths, plus an additional softer optically-thick component. The faintest three spectra were reproduced alternatively by a single Comptonized blackbody model with a relatively low ($\lesssim 0.8$) optical depth. The estimated radius of the blackbody emission, including seed photons for the Comptonization, was $10 \pm 2$ km at a 0.8--100 keV luminosity of $2.4\times 10^{36}$ erg s$^{-1}$ (the 2nd to the 4th observations). In contrast, it decreased to $7 \pm 1$ km and further to $3 \pm 1$ km, at a luminosity of $(4.8-5.2)\times 10^{35}$ erg s$^{-1}$ (the 5th observation) and $\sim 2\times 10^{34}$ erg s$^{-1}$ (the 6th and 7th), respectively, regardless of the above model ambiguity. This can be taken as evidence for the emergence of a weak magnetosphere of the neutron star.
Characterization of a Small Supernumerary Marker Chromosome Derived from Xq28 and 14q11.2 Detected Prenatally
Aikou Okamoto,Akihiro Hasegawa,Hiroaki Aoki,Kazuhiro Kajiwara,Masahisa Kobayashi,Osamu Samura,Taisuke Sato,Tomona Matsuoka,Yuka Inage,Yuki Ito
- , 2018, DOI: 10.1155/2018/2875241
Abstract:
Inventories and Mixed Duopoly with State-Owned and Labor-Managed Firms  [PDF]
Kazuhiro Ohnishi
iBusiness (IB) , 2010, DOI: 10.4236/ib.2010.22014
Abstract: This paper considers a two-period mixed market model in which a state-owned firm and a labor-managed firm are allowed to hold inventories as a strategic device. The paper then shows that the equilibrium in the second period occurs at the Stackelberg point where the state-owned firm is the leader.
Price-Setting Mixed Duopoly Models with Complementary Goods  [PDF]
Kazuhiro Ohnishi
Modern Economy (ME) , 2010, DOI: 10.4236/me.2010.11003
Abstract: This paper considers domestic (resp. international) Bertrand mixed duopoly competition in which a state-owned welfare-maximizing public firm and a domestic (resp. foreign) profit-maximizing private firm produce complementary goods. The main purpose of the paper is to present and to compare the equilibrium outcomes of the two mixed duopoly models.
Partial Privatization in Price-Setting Mixed Duopolies with Complementary Goods  [PDF]
Kazuhiro Ohnishi
Modern Economy (ME) , 2011, DOI: 10.4236/me.2011.21007
Abstract: We consider a domestic (resp. international) mixed duopoly model in which a domestic public firm and a domestic (resp. foreign) private firm produce complementary goods. First, the domestic government chooses the level of privatization to maximize domestic social welfare. Second, observing the level of privatization, the firms simultaneously and independently choose prices. We present the equilibrium outcomes of the two mixed duopoly models and shows that our result is in marked contrast to that of the price-setting mixed du-opoly model with substitute goods.
Labor Investment in a New International Mixed Market  [PDF]
Kazuhiro Ohnishi
Technology and Investment (TI) , 2010, DOI: 10.4236/ti.2010.12017
Abstract: This paper considers a continuous-time dynamic mixed market model of labor investment decisions of a domestic public firm and a foreign private firm. The paper studies the optimal levels of preemptive investment for the long-run structure of the international mixed market. It is then demonstrated that there are no perfect equilibria in which neither firm invests to its steady-state reaction curve.
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