Contribution of Invariant Natural Killer T Cells to the Clearance of Pseudomonas aeruginosa from Skin Wounds

Chronic infections are considered one of the most severe problems in skin wounds, and bacteria are present in over 90% of chronic wounds. Pseudomonas aeruginosa is frequently isolated from chronic wounds and is thought to be a cause of delayed wound healing. Invariant natural killer T (iNKT) cells, unique lymphocytes with a potent regulatory ability in various inflammatory responses, accelerate the wound healing process. In the present study, we investigated the contribution of iNKT cells in the host defense against P. aeruginosa inoculation at the wound sites. We analyzed the re-epithelialization, bacterial load, accumulation of leukocytes, and production of cytokines and antimicrobial peptides. In iNKT cell–deficient (Jα18KO) mice, re-epithelialization was significantly decreased, and the number of live colonies was significantly increased, when compared with those in wild-type (WT) mice on day 7. IL-17A, and IL-22 production was significantly lower in Jα18KO mice than in WT mice on day 5. Furthermore, the administration of α-galactosylceramide (α-GalCer), a specific activator of iNKT cells, led to enhanced host protection, as shown by reduced bacterial load, and to increased production of IL-22, IL-23, and S100A9 compared that of with WT mice. These results suggest that iNKT cells promote P. aeruginosa clearance during skin wound healing.     

Design,Synthesis, and Biological Evaluation of Lysine Demethylase 5 C Degraders

Lysine demethylase 5 C (KDM5C) controls epigenetic gene expression and is attracting great interest in the field of chemical epigenetics. KDM5C has emerged as a therapeutic target for anti-prostate cancer agents, and recently we identified triazole 1 as an inhibitor of KDM5C. Compound 1 exhibited highly potent KDM5C-inhibitory activity in in vitro enzyme assays, but did not show strong anticancer effects. Therefore, a different approach is needed for the development of anticancer agents targeting KDM5C. Here, we attempted to identify KDM5C degraders by focusing on a protein-knockdown strategy. Compound 3 b , which was designed based on compound 1 , degraded KDM5C and inhibited the growth of prostate cancer PC-3 cells more strongly than compound 1 . These findings suggest that KDM5C degraders are more effective as anticancer agents than compounds that only inhibit the catalytic activity of KDM5C.     

The Fe3O4-formation by the ‘Ferrite Process’: Oxidation of the reactive Fe(OH)2 suspension induced by sucrose

A mechanism of the formation of small particles of iron oxide in the wastewater treatment system ‘Ferrite Process’ is described. Small particles consisting of the Fe₃O₄−γ-Fe₂O₃ solid solution having slightly higher content of γ-Fe₂O₃ were obtained by the aerial oxidation of Fe(OH)₂ suspension in the presence of a weak dispersing reagent, sucrose, at pH 9.0. The increase of γ-Fe₂O₃ content is caused by a further oxidation of the Fe₃O₄ particles in the course of the reaction. The further oxidation was accelerated below 1000 Å of the particle size. When SO²⁻₄ ion coexisted with sucrose in the reaction medium, the further oxidation was reduced and the treatment of the wastewater was improved. At a temperature interval of 40°–65°C, the formation of α-FeOOH was completely inhibited by a small amount of sucrose and only Fe₃O₄ was obtained.     

Construction of Cellulose Binding Domain Fusion FMN-Dependent NADH-Azoreductase and Glucose 1-Dehydrogenase for the Development of Flow Injection Analysis with Fusion Enzymes Immobilized on Cellulose

The cellulose binding domain (CBD) of cellulosome-integrating protein A from Clostridium thermocellum NBRC 103400 was genetically fused to FMN-dependent NADH-azoreductase (AZR) and glucose 1-dehydrogenase (GDH) from Bacillus subtilis. The fusion enzymes, AZR-CBD and CBD-GDH, were expressed in Escherichia coli Rosetta-gami B (DE3). The enzymes were purified from cell-free extracts, and the specific activity of AZR-CBD was 15.1 U/mg and that of CBD-GDH was 22.6 U/mg. AZR-CBD and CBD-GDH bound strongly to 0.5 % swollen cellulose at approximately 95 and 98 % of the initial protein amounts, respectively. After immobilization onto the swollen cellulose, AZR-CBD and CBD-GDH retained their catalytic activity. Both enzymes bound weakly to 0.5 % microcrystalline cellulose, but the addition of a high concentration of microcrystalline cellulose (10 %) improved the binding rate of both enzymes. A reactor for flow injection analysis was filled with microcrystalline cellulose-immobilized AZR-CBD and CBD-GDH. This flow injection analysis system was successfully applied for the determination of glucose, and a linear calibration curve was observed in the range of approximately 0.16–2.5 mM glucose, with a correlation coefficient, r, of 0.998.     

A design and simulation tool for ground source heat pump system using energy piles with large diameter

Calculation of the underground temperature resulting from heat injection/extraction into/from ground heat exchangers (GHEXs) with hourly variation is one of the most noteworthy challenges to address when simulating and designing a ground source heat pump (GSHP). In order to overcome this challenge, the authors introduce a method to calculate the underground temperature, by considering heat injection/extraction into/from GHEXs with hourly variation. The method applies the superposition of the infinite cylindrical source (ICS) solution and the infinite line source (ILS) solution to calculate the temperature change due to heat injection/extraction into/from the considered GHEX and other neighboring GHEXs, respectively. The calculation method also considers heat injection/extraction from GHEXs with different heat injection/extraction rates and is able to accommodate GHEXs with large diameters such as energy piles. The calculation method was evaluated by applying it to calculate the temperature variation of the heat carrier fluid in a GSHP system with energy piles.     

Euphorbia tirucalli β-Amyrin Synthase: Critical Roles of Steric Sizes at Val483 and Met729 and the CH–π Interaction between Val483 and Trp534 for Catalytic Action

The functions of Val483, Trp534, and Met729 in Euphorbia tirucalli β-amyrin synthase were revealed by comparing the enzyme activities of site-directed mutants against that of the wild type. The Gly and Ala variants with a smaller bulk size at position 483 predominantly afforded monocyclic camelliol C, which suggested that the orientation of the (3S)-2,3-oxidosqualene substrate was not appropriately arranged in the reaction cavity as a result of the decreased bulk size, leading to failure of its normal folding into the chair–chair–chair–boat–boat conformation. The Ile variant, with a somewhat larger bulk, afforded β-amyrin as the dominant product. Intriguingly, various variants of Trp534 exhibited significantly decreased enzymatic activities and provided no aberrantly cyclized products, although the aromatic Phe and Tyr residues were incorporated and the steric sizes of the aliphatic residues were altered. Therefore, the Trp534 residue does not stabilize the transient cation through a cation–π interaction. Furthermore, the Trp residue, with the largest steric bulk among all natural amino acids, is essential for high enzymatic activity. Robust CH–π complexation between the Val483 and Trp534 residues is proposed herein. Altering the steric bulk at the Met729 position afforded the pentacyclic skeletons. Thus, Met729 is positioned at the E-ring formation site. More detailed insights into the functions of the Val483, Trp534, and Met729 residues are provided by homology modeling.     

Integrated reproduction of human motion components by motion copying system

Currently, the development of leading‐edge technology for recording and loading human motion on the basis of haptic information is required in the fields of manufacturing and human support. Human movement is an assembly of motion components. Since human movements should be supported by a robot in real time, it is necessary to integrate the motion components that were saved earlier. Once such motion integration is realized, future technology for use in daily human life can be developed. This paper proposes the integrated reproduction of the decomposed components of human motion by using a motion copying system. This system is the key technology for the realization of the acquisition, saving, and reproduction of real‐world haptic information. By using the proposed method, it is possible not only to achieve expert skill acquisition, skill transfer to robots, and power assist for each motion component, but also to open up new areas of applications. © 2012 Wiley Periodicals, Inc. Electr Eng Jpn, 181(1): 28–35, 2012; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/eej.21263     

Wind Force Coefficients for Designing Porous Canopy Roofs

Wind force coefficients for designing porous canopy roofs have been investigated based on a series of wind tunnel experiments. Gable, troughed and mono-sloped roofs were tested. The roof models were made of 0.5 mm thick perforated duralumin plates, the porosity of which was changed from 0 to about 0.4. Overall aerodynamic forces and moments acting on the roof model were measured in a turbulent boundary layer with a six-component force balance for various wind directions. The results indicate that the wind loads on canopy roofs generally decrease with an increase in porosity of the roof. Assuming that the roof is rigid and supported by the four corner columns with no walls, the axial forces induced in the columns are regarded as the most important load effect for discussing the design wind loads. Two loading patterns causing the maximum tension and compression in the columns are considered. Based on a combination of the lift and moment coefficients, the design wind force coefficients on the windward and leeward halves of the roof are presented for the two loading patterns as a function of the roof pitch and porosity. The effect of porosity is taken into account as a reduction factor of the wind loads.