The Functions of Metallothionein and ZIP and ZnT Transporters: An Overview and Perspective

Around 3000 proteins are thought to bind zinc in vivo, which corresponds to ~10% of the human proteome. Zinc plays a pivotal role as a structural, catalytic, and signaling component that functions in numerous physiological processes. It is more widely used as a structural element in proteins than any other transition metal ion, is a catalytic component of many enzymes, and acts as a cellular signaling mediator. Thus, it is expected that zinc metabolism and homeostasis have sophisticated regulation, and elucidating the underlying molecular basis of this is essential to understanding zinc functions in cellular physiology and pathogenesis. In recent decades, an increasing amount of evidence has uncovered critical roles of a number of proteins in zinc metabolism and homeostasis through influxing, chelating, sequestrating, coordinating, releasing, and effluxing zinc. Metallothioneins (MT) and Zrt- and Irt-like proteins (ZIP) and Zn transporters (ZnT) are the proteins primarily involved in these processes, and their malfunction has been implicated in a number of inherited diseases such as acrodermatitis enteropathica. The present review updates our current understanding of the biological functions of MTs and ZIP and ZnT transporters from several new perspectives.     

Mechanical role of reinforcement in seismic behavior of steel-strip reinforced earth wall

In the current design practices of steel-strip reinforced earth walls (SSREWs), the length of the reinforcing material is determined based on the equilibrium between the reinforcement tension and the earth pressure acting on the wall. Here, the resistance of the reinforcing material laid in the active failure zone (AFZ) is not considered. Moreover, the mechanical role of the reinforcing material against the integrity of the SSREW has not been sufficiently verified. Regarding the seismic stability of SSREW, although it is investigated by treating the entire reinforced earth wall as a rigid body, this inspection method is for gravity-retaining walls, and the difference in the seismic behavior between the SSREW and the rigid body is not clear. In this study, therefore, dynamic centrifuge model tests on 6 types of SSREWs were conducted to clarify the following items: (1) the basic earthquake behavior of a SSREW, (2) the mechanical role of the reinforcing material laid in the AFZ and (3) the mechanical role of the reinforcing material against the integrity of the SSREW. The results indicated that the reinforcing material laid in the AFZ can restrain the amount of deformation of the wall during earthquakes. Furthermore, the more stable the AFZ is, the smaller the maximum wall displacement will be.     

Collision response for deformable models based on hertz's contact theory

This paper presents an animation technique of collision response for deformable models defined as a spring-mass-damper system. Our approach is to calculate impulsive forces to prevent penetration of colliding objects by means of Hertz's contact theory. With this theory, contact duration and deformation in collision are obtained according to physical properties such as mass and elasticity. Animators, therefore, can represent the differences between materials such as rubber and steel in the sequence of collision animations with a few intuitive physical parameters. We also describe a deformation mapping technique which reduces the computational time of dynamic analysis and realizes the effect of global deformation.     

Molecular breeding of cellulolytic microbes, plants, and animals for biomass utilization

Cellulosic materials are the major components of fibrous biomass produced as a result of photosynthesis and are considered as a reservoir of solar energy and organic materials. In order to cope with the problems of food and energy shortages expected in the near future, biotechnologists are encouraged to develop new technologies for the more effective utilization of the world's sustainable resources, i.e., biomass. One way is to engineer microorganisms and animals with the capability of digesting and utilizing cellulosic materials, and plants which can be easily degraded by cellulolytic enzymes. In this article, we summarize recent studies on the molecular breeding of cellulolytic organisms for biomass utilization along with some considerations regarding cellulolytic enzymes.     

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.     

Cloning, sequencing and expression of an alpha-L-arabinofuranosidase from Aspergillus sojae

The arabinofuranosidase gene was cloned from the cDNA of Aspergillus sojae. It was found to contain an open reading frame composed of 984 base pairs (bp) and to encode 328 amino acid residues (aa). The cDNA sequence suggested that the mature enzyme is preceded by a 26-aa signal sequence and the molecular mass was predicted to be 32,749 Da. The A. sojae arabinofuranosidase consists of a single catalytic domain; it does not have a specific substrate-binding domain such as the xylan-binding domain reported in an arabinofuranosidase from Streptomyces lividans (Vincent, P. et al.: Biochem. J., 322, 845-852, 1997). The deduced amino acid sequence of the catalytic domain of the mature enzyme exhibits extensive identity with the catalytic domains of Streptomyces coelicolor (74%), Aspergillus niger (75%), S. lividans (74%), and Aspergillus tubingensis (75%), which are enzymes that belong to family 62 of the glycosyl hydrolases. The cloned AFdase gene was expressed in Escherichia coli BL21 (DE3) pLysS as a cellulose-binding domain tag fusion protein. The specific activity of the purified recombinant enzyme was 18.6 units/mg protein, which is one-fourth that of the enzyme purified from a solid-state culture of A. sojae.     

Synergistic Extraction of Am(III) and Eu(III) by Tris(2‐pyridylmethyl)Amine with Various Anions in 1,2‐Dichloroethane

The extraction equilibria of Am(III) and Eu(III) by using a tripodal ligand, tris(2‐pyridylmethyl)amine (tpa), with various lipophilic anions have been investigated. The extractability of both Am(III) and Eu(III) was increased by the combination of tpa and counteranions due to a synergistic effect. The separation factors between Am(III) and Eu(III) were also increased from 7.6 to 49 by the combination of counteranions and organic solvents. The extraction equilibria of Am(III) and Eu(III) with tpa in 1,2‐dichloroethane were determined by slope analysis. It was found that three anions and one molecule of the ligand coordinated to Am(III) and Eu(III) was extracted regardless of the anions.     

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).