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.     

Development of Fluorogenic Probes for Rapid High-Contrast Imaging of Transient Nuclear Localization of Sirtuin 3

Protein labeling using fluorogenic probes enables the facile visualization of proteins of interest. Herein, we report new fluorogenic probes consisting of a rationally designed coumarin ligand for the live-cell fluorogenic labeling of the photoactive yellow protein (PYP)-tag. On the basis of the photochemical mechanisms of coumarin and the probe–tag interactions, we introduced a hydroxy group into an environment-sensitive coumarin ligand to modulate its spectroscopic properties and increase the labeling reaction rate. The resulting probe had a higher labeling reaction rate constant and a greater fluorescence OFF–ON ratio than any previously developed PYP-tag labeling probe. The probe enabled the fluorogenic labeling of intracellular proteins within minutes. Furthermore, we used our probe to investigate the localization of sirtuin 3 (SIRT3), a mitochondrial deacetylase. Although the nuclear localization of SIRT3 has been controversial, this transient nuclear localization was clearly captured by the rapid, high-contrast imaging enabled by our probe.     

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.     

Force control of twisted and coiled polymer actuators via active control of electrical heating and forced convective liquid cooling

For decades, numerous artificial muscles have been proposed in order to implement beneficial features of biological muscles into robotics. Unfortunately, traditional artificial muscles experienced difficulties in imitating properties of the biological muscles due to mechanical and control issues. Recently, twisted and coiled polymer actuators (TCP) have been shown to produce large mechanical power via thermal stimulations and strong linearity. In this paper, a high-performance TCP thermally cycled by electrical heating and forced convective liquid cooling is designed and associated control algorithms are presented. We elaborate the model of the TCP that is simple, yet provides insight into how the electrical heating and the forced convective liquid cooling contribute to the TCP actuation. The proposed model is verified by experimental studies. Based on the proposed model, we design a feedforward–feedback controller and switching laws, which actively control the TCP in both the heating and cooling cycles. Furthermore, we extend our control methodology to agonist–antagonist TCPs. From the experimental studies, the proposed method is shown to be effective in both single TCP and antagonistic TCPs.     

Hydrogen production from methane steam reforming: parametric and gradient based optimization of a Pd-based membrane reactor

In this work three mathematical models for methane steam reforming in membrane reactors were developed. The first one is a steady state, non isothermal, non isobaric and one dimensional model derived from material and energy balances and validated using experimental data from the literature. It is referred as full model. The influence of two different intrinsic kinetics available, as well as, the influence of five important parameters on methane conversion (X_CH4_{\mathrm{CH}_{4}}) and hydrogen recovery (Y_H2_{\mathrm{H}_{2}}) were parametrically evaluated through simulations. The second model, referred as meta-model, was obtained though the response surface technique. This meta-model was included into a constrained optimization problem solved using NPSOL. The third model, referred as a simplified model, takes into account only mass balances from the full model. Using this model, a gradient based method (DIRCOL) was used to perform the optimization of the sum of methane conversion and hydrogen recovery. High methane conversions and hydrogen recoveries were reached through these methodologies.     

Model Test and Consolidation Analysis of Failure of a Loose Sandy Embankment Dam During Seepage

Failures of earth dam embankments and river dikes, which are constructed of sandy soils with low dry densities, have been observed to occur during rising of water levels. In this paper, a large-scale physical model test was conducted in order to investigate the behavior of a small dam embankment as water levels rose. The test results were simulated by use of a consolidation analysis method coupled with an elastoplastic model for unsaturated soils. All parameters used in the simulation were obtained from element tests; oedometer, triaxial compression, soil water retention, and permeability tests. First, in order to verify the parameters identified, direct shear tests were simulated using the consolidation analysis method; results of the simulation and the tests were consistent. From the embankment model test it was found that the crest of the embankment moved upstream at the first stage of impounding and then moved back. After a seepage surface appeared on the downstream slope, tension cracks occurred on the downstream slope, and sliding occurred through the crack and the downstream toe. The results of the simulation were consistent with those of the model test. This consolidation analysis method could be used to simulate the complex deformations induced by saturation collapse and shear strains and even failure behavior. Old embankments constructed with loose densities might have histories in which cracks occurred on the downstream slopes when the reservoir water level rose, and their stabilities might have decreased.     

Supercritical fluid‐assisted electroless metal plating onto aramid films: The influence of thermal treatment

Although the production of electro‐conductive aramid fibers is efficient, the method needs to be modified before it can be applied to aramid films. Whereas impregnation of an aramid film with a metal complex using supercritical CO₂ is achievable, the relatively low adhesion strength of the metal layer applied using electroless copper plating is problematic. To solve this problem, thermal treatment was conducted before, after, or both before and after electroless plating. The rationale for using thermal treatment to improve the adhesiveness of the plated layer was based on the findings that (1) an aramid film contains a significant amount of water (about 3.5 % w.o.f.), which might have a negative impact on adhesion; and (2) because an impregnated metal complex liberates metal catalyst by thermal decomposition during impregnation, a supplementary thermal action might liberate more catalyst and thereby improve adhesion. We found that thermal treatment improved adhesion of the metal layer to the aramid film. Moreover, we discovered that with respect to electroless copper plating, a short time‐lag was crucial to obtaining a thin and homogeneous metal layer with strong adhesion. In addition, we demonstrate the affinity of an aramid film for Pd(acac)₂. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2011