Nanoimprint with thin and uniform residual layer for various pattern densities

Pattern density variation is uncomfortable for nanoimprint lithography which uses a moldable material supplied as a thin film, because the variation of pattern density causes variations of residual layer thickness reflecting on the local pattern density. To solve the problem, a new type of mold “capacity-equalized mold”, which has constant averaged depth regardless of pattern density, was fabricated and the structure of the mold was inspected. UV nanoimprint was then carried out using the mold and thickness and uniformity of the residual layer were investigated. An average thickness of 33.2 nm with a standard deviation of 3.4 nm was obtained for the mold pattern layout with a pattern density of from 0.25 to 0.75. It was found that a standard deviation of 1.2 nm was achieved for pattern density variation of from 0.33 to 0.67 by excluding artifacts.     

Building maps of local apparent conductivity of the epicardium with a 2-D electrophysiological model of the heart

In this paper, we address the problem of estimating the parameters of an electrophysiological model of the heart from a set of electrical recordings. The chosen model is the reaction-diffusion model on the transmembrane potential proposed by Aliev and Panfilov. For this model of the transmembrane, we estimate a local apparent two-dimensional conductivity from a measured depolarization time distribution. First, we perform an initial adjustment including the choice of initial conditions and of a set of global parameters. We then propose a local estimation by minimizing the quadratic error between the depolarization time computed by the model and the measures. As a first step we address the problem on the epicardial surface in the case of an isotropic version of the Aliev and Panfilov model. The minimization is performed using Brent method without computing the derivative of the error. The feasibility of the approach is demonstrated on synthetic electrophysiological measurements. A proof of concept is obtained on real electrophysiological measures of normal and infarcted canine hearts.     

A Real-coded Genetic Algorithm Applied to Optimum Design of a Low Solidity Vaned Diffuser for Diffuser Pinup

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Influence of side chain length on fluorescence intensity of ROMP-based polymeric nanoparticles and their tumor specificity in in-vivo tumor imaging

In this study, amphiphilic brush-like copolymers conjugated with short alkyl or long polymeric chains of various lengths are synthesized using ring-opening metathesis polymerization (ROMP) of substituted norbornadiene monomers followed by chemical transformations. These amphiphilic copolymers form spherical self-assemblies in aqueous media with diameters of 132-244 nm. The low critical aggregation concentration of these assemblies (2.5 × 10(-3) -1.4 × 10(-5) g/L) indicates that they are quite stable in dilute conditions. An appropriate length of polymer side chain that conjugates the polymer backbone with a hydrophobic ICG (indocyanine green) moiety enhanced the fluorescence intensities of these self-assemblies in aqueous solution as well as in tumor-bearing mice. A longer side chain conjugated with tumor targeting agents could significantly affect the tumor specificity of self-assemblies to a greater extent. The self-assemblies bearing hydrophilic tumor targeting agents, such as a glucosamine molecule and a cyclic RGD (arginine-glycine-asparatic acid) peptide, accumulated in tumor tissues with high selectivity, while those having a hydrophobic targeting agent, such as folate moieties, accumulated in tumor sites with low selectivity. The results demonstrated here unambiguously indicate that the fluorescence intensity and tumor specificity of self-assemblies are strongly affected by the length of side chains that conjugate with dyes and targeting agents.     

Biomimetic assembly of proteins into microcapsules on oil-in-water droplets with structural reinforcement via biomolecular-recognition-based cross-linking of surface peptides

By mimicking the stabilization of bacterial membranes with S-layer proteins, a novel process to fabricate highly stable protein microcapsules is introduced. In this strategy, engineered collagen peptides with site-specific biotinylation are assembled into microcapsules on the oil-in-water droplets, and the resulting microcapsules are reinforced by biomolecular-recognition-based cross-linking with the protein. Furthermore the microcapsules are shown to be versatile scaffolds for developing functionalized hierarchical colloidosomes for important biotechnological applications.     

Enhancement of thermostability of kojibiose phosphorylase from Thermoanaerobacter brockii ATCC35047 by random mutagenesis

Random mutation by error-prone PCR was introduced into kojibiose phosphorylase from Thermoanaerobacter brockii ATCC35047. One thermostable mutant enzyme, D513N, was isolated. The D513N mutant enzyme showed an optimum temperature of 67.5-70 degrees C (the wild type, 65 degrees C), and thermostability up to 67.5 degrees C (the wild type, up to 60 degrees C). The half-lives of D513N were estimated to be 135 h at 60 degrees C, 110 min at 70 degrees C and 6 min at 75 degrees C, respectively. They were about 1.6-fold, 7-fold and 6-fold longer than those of the wild-type enzyme, respectively.     

Reconstruction of cartilage tissue using scaffold-free organoid culture technique

We applied the scaffold-free culture method to chondrocytes and attempted to reconstitute articular cartilage grafts. Primary rat costal chondrocytes were immobilized into hollow fibers by centrifugation at a density of 3 x 10(8) cells/cm(3) to induce the formation of cylindrical-shaped multicellular aggregates (organoids) and cultured for one month. The organoids were evaluated by histological and gene expression analyses. Chondrocytes formed cylindrical organoids in hollow fibers (HFs). Histochemical analysis revealed the accumulation of a cartilage extracellular matrix (collagen and proteoglycan) around cells in the lumen of HFs with culture time, forming a low-cellular-density tissue similar to native cartilage by day 28. Furthermore, in contrast to that in traditional monolayer culture, the organoid maintained the gene expression of the cartilage extracellular matrix (type II collagen, aggrecan) for one month of culture. In conclusion, our organoid formation method was effective in producing a cartilage-like tissue. This result suggests that the technique may be applicable to the development of an articular cartilage graft.     

Utilization of adzuki bean extract as a natural antioxidant in cured and uncured cooked pork sausages

A commercial adzuki bean extract (AE) was evaluated for antioxidant effectiveness in cured and uncured cooked pork sausages. TBARS values, instrumental color evaluation and sensory panel scores were assessed. For uncured sausages, AE at 0.2% was equally effective as 0.1% butylated hydroxytoluene (BHT) in reducing TBARS values. Similarly, AE at 0.2% significantly (P<0.01) reduced the TBARS in cured sausages. Incorporation of 0.2% AE into sausages produced higher (P<0.05) CIE lab color a* value and lower (P<0.05) L* and b* values. Sensory panels did not detect any difference in color, odor, taste, flavor, and overall acceptance in uncured pork sausages with addition of 0.2% AE. However, there were adverse changes in the color and odor of cured sausages, even though the taste, flavor, and overall acceptance were similar. Therefore, the results suggest that AE is a potential antioxidant.     

Decomposition of extremely hard-to-degrade animal proteins by thermophilic bacteria

Hard-to-degrade animal proteins are ubiquitously present throughout animal bodies. Enormous numbers of these proteins generated in the meat industry are converted to industrial wastes, the disposal of which is tremendously difficult. Most hard-to-degrade animal proteins are currently disposed of by incineration; however, this method has ecological disadvantages in terms of an apparent energy loss and the production of a large amount of carbon dioxide. As a result, an innovative solution to these problems has been sought. In this review, we focus on the degradation of three hard-to-degrade animal proteins (extracellular matrix proteins, collagen in particular, keratin, and prion proteins) and discuss the decomposing capability of thermophilic bacteria. These proteins are strongly resistant to proteinases because of their structural features; therefore, new approaches employing bacterial proteases with strong activity and broad specificity are required for practical application.     

Effect of fermented milk prepared with two probiotic strains on Japanese cedar pollinosis in a double-blind placebo-controlled clinical study

There has been much interest in the potential of using probiotic bacteria for treating allergic diseases. A double-blind, placebo-controlled study was conducted to examine the effectiveness of Lactobacillus GG (LGG) and L. gasseri TMC0356 (TMC0356) in alleviating Japanese cedar pollinosis (JCP), a seasonal allergic rhinitis caused by Japanese cedar pollen. Fermented milk prepared with the tested bacteria or placebo yoghurt was administered to 40 subjects with a clinical history of JCP for 10 weeks. Subjective symptoms, self-care measures and blood samples were compared between the two groups. Peripheral blood mononuclear cells (PBMCs) were collected from seven patients with JCP and in vitro cytokine production by the isolated PBMCs was analysed in the presence of heat-killed lactic acid bacteria. Consumption of the fermented milk significantly decreased the mean symptom score for nasal blockage after 9 weeks (P<0.05) and mean symptom-medication scores after 9 and 10 weeks when compared with the placebo group (P<0.01 and P<0.05 respectively). The tested strains of lactic acid bacteria affected cytokine production by isolated PBMCs in vitro in a strain-dependent manner. LGG significantly inhibited IL-4 and IL-5 production by PBMCs stimulated by both Cry j 1 and PHA. TMC0356 only suppressed IL-5 production stimulated by PHA. The fermented milk prepared with LGG and TMC0356 might be beneficial in JCP because of its effect on nasal blockage. The effects of LGG and TMC0356 might arise at least partly from their specific down-regulation of the human Th2 immune response.