Metallocenium Ionic Liquids

正常造血と貧血4学

Composition and Antioxidant Activity of Rice Fermented with Saccharifying Organisms from Asian Countries

In recent years, more effective use of rice has become important because of an annual increase in surplus rice. We fermented rice in pure cultures of eight organisms (Aspergillus oryzae, Monaseus pilosus, Absidia corymbifera, Mucor circinelloides, Mucor racemosus, Rhizopus oligosporus, Rhizopus oryzae, and Saccharomycopsis fibuligera), which were isolated from molded rice and soybean products in Asian countries, and evaluated the composition and antioxidant activity of the products. Rice fermented with the two Rhizopus species had a high methanol extract yield, implying good fermentation properties. High saccharification and increased levels of total amino acids and total polyphenols were also found in Rhizopus-fermented rice samples. Ethyl acetate extracts of rice fermented with Ab. corymbifera and Mu. circinelloides had enhanced antioxidant activity compared to unfermented rice, and some fractions obtained from the extracts by high performance liquid chromatography exhibited high antioxidant activity. Based on these results, Ab. corymbifera, Mu. circinelloides, R. oligosporus, and R. oryzae are promising starter organisms for the development of new fermented rice products.ArticleFOOD SCIENCE AND TECHNOLOGY RESEARCH. 19(5):893-899 (2013)journal articl

Expression, Purification and Properties of Alanine Racemase from Thermus thermophillus HB8

An alanine racemase (EC 5.1.1.1) from an extreme thermophilic bacterium Thermus thermophilus HB8, was purified and characterized, and its gene was cloned. The cloned alanine racemase gene (alr) was expressed in Escherichia coli JM 109. The alr gene is composed of a 1080 bp and encoded a 360 amino acid, and was predicted to have a molecular weight of 38,596. The enzyme was purified by heat shock at 70°C for 10min and DEAE Toyopearl 650M column chromatography. The purified enzyme had an optimum pH9.0∼10.0 and an optimum temperature of 55°C∼60°C. Enzyme activity was retained 100% after incubation of the enzyme at 70°C for 10min. Alanine racemase from Thermus thermophilus HB8 is a monomeric enzyme with a molecular mass of 39 kDa.高度好熱性細菌 Thermus thermophilus HB8 由来アラニンラセマーゼ遺伝子を大腸菌中にクローニングし、発現させた後に、精製及び性質検討を行った。alr遺伝子は1080bpからなり360アミノ酸残基 HB8をコードしていたので、本酵素は38,596Daの分子量であると予想された。alr遺伝子の [ G + C ] 含量は、72%であり、Tm値は98.8℃であった。T.thermophilus HB8由来アラニンセマーゼを中等度好熱性細菌 Geobacillus stearothermophilus 及び赤痢菌 Shigella sonnei 由来アラニンラセマーゼと一次配列の比較をしたところ、G.stearothermophilus 由来のものと33%、赤痢菌由来の酵素を28%の相同性を示した。T.thermophilus HB8 由来アラニンラセマーゼを、70℃で10分間の熱処理後、DEAE-トーヨーパール陰イオン交換カラム等により精製した。精製酵素の最適温度は、d-アラニンからl-アラニンへの反応で55℃、l-アラニンからd-アラニンへの反応では60℃であり、最適pHは、9.0〜10.0であった。また、70℃で30分インキュベーションを行った後にも、活性の低下は見受けられず耐熱性を示した。更に、本酵素は分子量38,000モノマー酵素であると推定され、その反応機構に興味が持たれる

Development of Safety Measures of Bicycle Trafflc by Observation wffh Deep-Leamlng, Drive Recorder Data, Probe Blcycle wlth LIDAR, and Connected Simulators

This research outlines the development of evaluating safety measures for bicycle traffic using state-of-the-art technology, which was started since 2020 as a four-year project. The project is funded by the Commission on Advanced Road Technology in the Ministry of Land, Infrastructure, Transport and Tourism(MLIT). While Japan has a high bicycle modal share of 12% (2010), bicycle-related fatalities are relatively high among other countries in the IRTAD database (2019). Under these circumstances, since 2007, various measures for bicycle traffic measures have been implemented to improve the safe bicycle traffic environment, including the revision of the Road Traffic Act and the formulation of a national plan to promote bicycle use. However, serious accidents involving bicycles are remained in some specific cases. According to the government's traffic accident analysis results (2019), right-hook crash at signalized intersections are one of the most serious types of collision involving bicycles, along with accidents at unsignalized intersections involving vehicles turning left, rear-end collisions, and single vehicle accidents due to off-road deviation. In particular, proactive safety measures are required at signalized intersections along arterial roads, where electric personal mobility vehicles traveling at speeds of up to 20 km/h are expected to share with bicycles in the future. In order to evaluate safety measures for bicycle-vehicle crashes, this project set the following goals. 1) Identify factors influencing near-miss incidents and collisions through analysis of drive recorder data and accident statistical data. 2) Detailed analysis of traffic conditions from the cyclist's perspective using a probe bicycle equipped with a LiDAR sensor. 3) Development of an experimental environment using a connected simulator for evaluation of cooperative driving behavior. 4) Clarification of experimental conditions to evaluate different scenarios and conditions with and without intervention. 5) Proposal of effective interventions to improve crash cases based on experiments

Experimental observations on the optimal layout of orientation blocks for safe road crossing by the visually impaired

For people with visual impairments who face difficulties when crossing the road, in urban areas of Japan the infrastructure designed to provide an indication of crossing direction and the curbstones at sidewalk-roadway boundaries often varies in reliability from one crossing to another. If anything, this promotes stress for users and is an issue for which improvement is urgently needed. The authors have proposed new orientation blocks to be installed at crosswalk entrances as a means of more accurately indicating to people with visual impairments the trajectory to follow when crossing the road, and in prior research have derived desirable specifications for the profile of these blocks and their position relative to tactile walking surface indicators (TWSI). For this paper, in order to examine in greater detail the desirable position of orientation blocks relative to TWSI, the authors conducted an experiment using totally blind subjects to evaluate conditions on a 10 m walk that simulated an actual crossing. The results, based on observations of the trajectories walked by participants in the experiment and interviews eliciting their subjective evaluations, showed that separating orientation blocks and blister tactile blocks by about 8–12 cm is effective in constraining lateral deviation at a point 5 m from the start of crossing and that an 8 cm separation was desirable in order to maintain an effective reduction of mental stress while crossing

Inhibition of IgE-dependent Mouse Triphasic Cutaneous Reaction by a Boiling Water Fraction Separated from Mycelium of Phellinus linteus

Phellinus linteus, a mushroom, contains constituents that exhibit potent antitumor effects through activating immune cells. Recently, anti-inflammatory and anti-allergic properties of P. linteus extracts have also been implicated. In the present study, therefore, we separated the constituents of mycelium of P. linteus into five fractions—chloroform-soluble (CF), ethyl acetate-soluble (EA), methanol-soluble (AE), water-soluble (WA) and boiling water-soluble (BW) fractions—and examined their suppressive effects on the IgE-dependent mouse triphasic cutaneous reaction. The triphasic reaction was induced in the ear of BALB/c mice passively sensitized with anti-dinitrophenol IgE by painting with 2,4-dinitrofluorobenzene 24 h later. Ear swelling appeared triphasically with peak responses at 1 h, 24 h and 8 days after the challenge. ME, WA and BW given orally at a dose of 100 mg kg(−1) significantly inhibited the first and second phase ear swelling, and BW also inhibited the third phase response. CF only inhibited the second phase. The inhibition by BW was the most potent and almost dose-dependent at doses of 30–300 mg kg(−1). BW also inhibited vascular permeability increase caused by passive cutaneous anaphylaxis and histamine, and ear swelling caused by tumor necrosis factor-α. In contrast, BW apparently potentiated the production of interleukin-4 and interferon-γ from anti-CD3-stimulated mouse splenocytes. These results indicate that BW derived from mycelium of P. linteus contains some constituents with anti-allergic as well as immunopotentiating properties

骨欠損再建における培養細胞シートを併用した培養人工骨移植の有用性

AIM: To determine the effects of transplanting osteogenic matrix cell sheets and beta-tricalcium phosphate (TCP) constructs on bone formation in bone defects. METHODS: Osteogenic matrix cell sheets were prepared from bone marrow stromal cells (BMSCs), and a porous TCP ceramic was used as a scaffold. Three experimental groups were prepared, comprised of TCP scaffolds (1) seeded with BMSCs; (2) wrapped with osteogenic matrix cell sheets; or (3) both. Constructs were implanted into a femoral defect model in rats and bone growth was evaluated by radiography, histology, biochemistry, and mechanical testing after 8 wk. RESULTS: In bone defects, constructs implanted with cell sheets showed callus formation with segmental or continuous bone formation at 8 wk, in contrast to TCP seeded with BMSCs, which resulted in bone non-union. Wrapping TCP constructs with osteogenic matrix cell sheets increased their osteogenic potential and resulting bone formation, compared with conventional bone tissue engineering TCP scaffolds seeded with BMSCs. The compressive stiffness (mean ± SD) values were 225.0 ± 95.7, 30.0 ± 11.5, and 26.3 ± 10.6 MPa for BMSC/TCP/Sheet constructs with continuous bone formation, BMSC/TCP/Sheet constructs with segmental bone formation, and BMSC/TCP constructs, respectively. The compressive stiffness of BMSC/TCP/Sheet constructs with continuous bone formation was significantly higher than those with segmental bone formation and BMSC/TCP constructs. CONCLUSION: This technique is an improvement over current methods, such as TCP substitution, and is useful for hard tissue reconstruction and inducing earlier bone union in defects.博士（医学）・乙第1366号・平成27年11月27日Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved. This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license,which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/ licenses/by-nc/4.0

Field Survey of Flank Collapse and Run-up Heights due to 2018 Anak Krakatau Tsunami

Dataset: https://data.4tu.nl/articles/dataset/Bathymetry_data_underlying_the_publication_Field_survey_of_flank_collapse_and_run-up_heights_due_to_the_2018_Anak_Krakatau_Tsunami/1421561

Construction of an All-in-one Double-conditional shRNA Expression Vector

Gene silencing by RNA interference （RNAi） is widely used for assessing gene function. An important advance in the RNAi field was the discovery that plasmid-based RNAi can substitute for synthetic small interfering RNA in vitro and in vivo. However, constitutive and ubiquitous knockdown of gene expression by RNAi in mice can limit the scope of experiments because this process can lead to embryonic lethality, or result in compensatory overexpression of other genes such that no phenotypic abnormalities occur. Either way, analyses of the physiological roles of the gene of interest in adult mice are not possible. To overcome these limitations, we previously constructed a double-conditional short-hairpin RNA （shRNA） expression vector that can regulate shRNA expression in a spatio-temporal manner with a tetracycline-inducible floxed stuffer sequence selectively excised by application of Cre recombinase. In this study, we aimed to modify this vector to create an all-in-one vector that produces double-conditional transgenic mice through a single round of gene transfer to fertilized eggs. We added a coding region for nuclear localizing Cre （NCre） recombinase with a multi-cloning site for a cell-specific promoter into the double-conditional short-hairpin RNA （shRNA） expression vector that we previously constructed. Using Escherichia coli, we confirmed successful construction of the vector. First, we confirmed isopropyl-β-D-thiogalactopyranoside-induced expression of NCre recombinase through the lac operon as a specific promoter by western blotting. Second, we confirmed functional recombination of the floxed sequence of loxP-like TATA-lox by analysing restriction enzyme-digested fragments. This all-in-one double-conditional shRNA expression vector will be useful for reversible in vitro and in vivo knockdown of target gene expression, in target cells via promoter-specific expression of NCre, and at specific times by tetracycline application

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead