Fabrication of CaCO3-Coated Vesicles by Biomineralization and Their Application as Carriers of Drug Delivery Systems

We fabricated CaCO₃-coated vesicles as drug carriers that release their cargo under a weakly acidic condition. We designed and synthesized a peptide lipid containing the Val-His-Val-Glu-Val-Ser sequence as the hydrophilic part, and with two palmitoyl groups at the N-terminal as the anchor groups of the lipid bilayer membrane. Vesicles embedded with the peptide lipids were prepared. The CaCO₃ coating of the vesicle surface was performed by the mineralization induced by the embedded peptide lipid. The peptide lipid produced the mineral source, CO₃²⁻, for CaCO₃ mineralization through the hydrolysis of urea. We investigated the structure of the obtained CaCO₃-coated vesicles using transmission electron microscopy (TEM). The vesicles retained the spherical shapes, even in vacuo. Furthermore, the vesicles had inner spaces that acted as the drug cargo, as observed by the TEM tomographic analysis. The thickness of the CaCO₃ shell was estimated as ca. 20 nm. CaCO₃-coated vesicles containing hydrophobic or hydrophilic drugs were prepared, and the drug release properties were examined under various pH conditions. The mineralized CaCO₃ shell of the vesicle surface was dissolved under a weakly acidic condition, pH 6.0, such as in the neighborhood of cancer tissues. The degradation of the CaCO₃ shell induced an effective release of the drugs. Such behavior suggests potential of the CaCO₃-coated vesicles as carriers for cancer therapies.     

First nuclear transmutation of 237Np and 241Am by accelerator-driven system at Kyoto University Critical Assembly

This study demonstrates, for the first time, the principle of nuclear transmutation of minor actinide (MA) by the accelerator-driven system (ADS) through the injection of high-energy neutrons into the subcritical core at the Kyoto University Critical Assembly. The main objective of the experiments is to confirm fission reactions of neptunium-237 (²³⁷Np) and americium-241 (²⁴¹Am), and capture reactions of ²³⁷Np. Subcritical irradiation of ²³⁷Np and ²⁴¹Am foils is conducted in a hard spectrum core with the use of the back-to-back fission chamber that obtains simultaneously two signals from specially installed test (²³⁷Np or ²⁴¹Am) and reference (uranium-235) foils. The first nuclear transmutation of ²³⁷Np and ²⁴¹Am by ADS soundly implemented by combining the subcritical core and the 100 MeV proton accelerator, and the use of a lead-bismuth target, is conclusively demonstrated through the experimental results of fission and capture reaction events.     

Immobilization of alkaline phosphatase on magnetic particles by site-specific and covalent cross-linking catalyzed by microbial transglutaminase

Bacterial alkaline phosphatase (BAP) was site-specifically and covalently immobilized on magnetic particles (MPs) using the enzymatic reaction of microbial transglutaminase (MTG). Immobilization efficiency was affected by the chemical surface treatment of MPs and immobilized BAP exhibited more than 90% of the initial activity after 10 rounds of recycling.     

Detection of Leuconostoc strains at a meat processing plant using polymerase chain reaction

To simplify the labor-intensive conventional routine testing of samples to detect Leuconostoc at a meat processing plant, we developed polymerase chain reaction (PCR) primers specific for Leuconostoc from 16S rRNA gene sequences. These primers did not detect other common lactic acid bacteria such as Lactobacillus plantarum, Lact. sake, Lact. fermentum, Lact. acidophilus and Weissella viridescens. PCR with this primer detected all Leuconostoc species tested (Leu. mesenteroides subsp. mesenteroides, Leu. pseudomesenteroides, Leu. carnosum, Leu. lactic, Leu. citreum, Leu. amelibiosum, Leu. gelidum), except for Leu. fallax, and no other lactic acid bacteria on agarose gel electrophoresis. The method could identify areas contaminated with Leuconostoc in a large-scale industrial meat processing plant. Of 69 samples analyzed, 34 were positive for Leuconostoc according to the conventional culture method (isolation of LAB producing dextran) and PCR, whereas 29 were negative according to both. Six samples were culture-negative but positive by PCR. No false negative results were generated by PCR. The method is rapid and simple, is useful for routinely monitoring areas contaminated with Leuconostoc in meat processing plants, and could help to prevent the spoilage of meat products.     

The viscoelastic properties of soybean curd (tofu) as affected by soymilk concentration and type of coagulant

The viscoelastic properties of different types of tofu were investigated. Soymilk concentrations were 5, 6, 7, 8 and 9%. Coagulants used were 30 mm CaSO₄ or 30 mm glucono‐delta‐lactone (GDL). As the concentration of soymilk was increased viscosity and handling difficulties increased. A high concentration of soymilk in tofu gave a high break stress and produced hard tofu. The four‐element Burgers model fitted the creep behaviour and both viscous and elastic parameters could be acquired from model analysis, reflecting changes in elasticity and viscosity of tofu. The constant viscous parameter in the model increased with increasing soymilk concentration. The viscous parameters of viscoelastic materials like tofu gel, obtained from small deformation tests, seemed to correlate, to some extent, with the break stress obtained from large deformation tests. For hard tofu production increasing the soymilk concentration within a certain range and the partial replacement of calcium sulphate coagulant by GDL could be effective options.     

Package Design of Ready-to-Drink Coffee Beverages Based on Food <Emphasis Type="Italic">Kansei</Emphasis> Model—Effects of Straw and Cognition Terms on Consumer’s Pleasantness

To design a consumer-oriented package that complements the taste and aroma of ready-to-drink chilled-cup coffee beverages by using the food kansei model, the effects of the diameter and the color of drinking straws as well as the cognition terms of coffee on consumer sensory characteristics and preferences were investigated. Variance and factor analyses of the sensory scores for chilled-cup coffee with milk and sugar using straws of different diameters, as rated by consumer panelists, extracted two perceived factors (F1, contribution ratio 36.5%, and F2, 28.6%). A two-dimensional plot of the average F1 and F2 scores of 123 panelists showed that the perceived characteristics of the same taste and aroma varied according to the straw diameter. An image investigation of different straw colors and another sensory evaluation using “black,” “brown,” and “green” straws were also performed. A principal component analysis of the image data revealed that the sensory characteristics of coffee with milk and sugar were imaged from the straw color. The second evaluation suggested that the images of straw colors affected the sensory characteristics. In addition, cluster and multiple-comparison analyses of Internet research data from consumers regarding the cognition terms for coffee exhibited three clusters representing the cognitive characteristics of terms by consumers and by developers and the differences of attractiveness degree on the cognition terms due to the consumers’ personal attributes. These studies provide useful information for the development of packages of chilled-cup coffee beverages.     

Cloning and sequence analysis of a gene (aly PG) encoding poly(alpha-L-guluronate)lyase from Corynebacterium sp. strain ALY-1

The aly PG gene, coding for a poly alpha-l-guluronate lyase (PG lyase) of Corynebacterium strain ALY-1, was cloned and sequenced. The gene consists of 768 bp encoding a signal peptide of 32 amino acids and a mature protein of 224 amino acids. Two disulfide bond cross-linkages were found to be formed between Cys-4 and Cys-51 and between Cys-200 and Cys-206 in the native PG lyase molecule. The deduced amino acid sequence of the Corynebacterium sp. aly PG gene exhibited 29% homology toward that of the Klebsiella pneumoniae, subsp. aerogenes aly A gene, with two conserved regions (the amino acid sequences from Y-102 to M-110 and from Y-221 to Q-229).     

Graphitization of carbon fibre/ glassy carbon composites

Carbon fibre/glassy carbon composites were prepared by aligning unidirectionally in furfuryl alcohol condensate the PAN-based carbon fibres treated at different temperatures and with different degrees of stretching. The graphitization of the composites was found to start at the boundary between the fibres and glassy carbon matrix, and to proceed into the matrix. This is considered to be due to the stress accumulation at the boundary caused by a large shrinkage of the matrix. The carbon fibres remain nongraphitized even after a heat-treatment at 2800°C. The composites heat-treated at high temperature (2800°C) are found to show a high overall degree of graphitization, unexpected on basis of the known graphitization behavior of carbon fibres and of glassy carbon, and a high degree of uniaxial preferred orientation of crystallites.     

Mechano-electrochemistry of a passive surface using an in situ micro-indentation test

Depassivation–repassivation of iron surfaces in boric–borate solutions were investigated by using the micro-indentation test. A pair of current peaks due to repair of the passive film following rupture of the film were observed during a series of indenter drives, i.e., loading and unloading of the indenter. The shape of the current peak depended on environmental conditions (conductivity and pH of the solution) and substrate conditions (mechanical processing history, alloyed element) as well as indentation conditions (repetition, maximum depth, and maximum load). Plastic deformation of the surface was accompanied by surface depassivation, while no depassivation occurred during the elastic deformation, indicating that the passive film on iron has a ductile property. The solution conditions did not affect the scale of depassivation but affected the rate of repassivation. Dislocations in the substrate made surface depassivation difficult but enhanced reactivity during the repassivation. The test also revealed that type-312L stainless steel has high corrosion resistance in a concentrated NaCl solution.     

Deposition of Lead Phosphate by Lead-Tolerant Bacteria Isolated from Fresh Water near an Abandoned Mine

Specialist bacteria can synthesize nanoparticles from various metal ions in solution. Metal recovery with high efficiency can be achieved by metal-tolerant microorganisms that proliferate in a concentrated metal solution. In this study, we isolated bacteria (Pseudomonas sp. strain KKY-29) from a bacterial library collected from water near an abandoned mine in Komatsu City, Ishikawa Prefecture, Japan. KKY-29 was maintained in nutrient medium with lead acetate and synthesized hydrocerussite and pyromorphite nanoparticles inside the cell; KKY-29 also survived nanoparticle synthesis. Quantitative PCR analysis of genes related to phosphate metabolism showed that KKY-29 decomposed organic phosphorus to synthesize lead phosphate. KKY-29 also deposited various metal ions and synthesized metal nanoparticles when incubated in various metal salt solutions other than lead. The present study considers the development of biotechnology to recover lead as an economically valuable material.