Discrimination of psychoactive fungi (commonly called "magic mushrooms") based on the DNA sequence of the internal transcribed spacer region

'Magic mushrooms' (MMs) are psychoactive fungi containing the hallucinogenic compounds, psilocin (1) and psilocybin (2). Since June 6, 2002, these fungi have been regulated by the Narcotics and Psychotropics Control Law in Japan. Because there are many kinds of MMs and they are sold even as dry powders in local markets, it is very difficult to identify the original species of the MMs by morphological observation. Therefore, we investigated the internal transcribed spacer (ITS) region in the ribosomal RNA gene of MMs obtained in Japanese markets to classify them by a genetic approach. Based on the size and nucleotide sequence of the ITS region amplified by PCR, tested MMs were classified into 6 groups. Furthermore, a comparison of the DNA sequences of the MMs with those of authentic samples or with those found in the databases (GenBank, EMBL and DDBJ) made it possible to identify the species of tested MMs. Analysis by LC revealed that psilocin (1) was contained at the highest level in Panaeolus cyanescens among the MMs, but was absent in the Amanita species.     

DNA and chemical analyses of commercial fly agaric-related products

Since June 6, 2002, psilocin and psilocybin-containing fungi (commonly called "magic mushrooms") have been regulated by the Narcotics and Psychotropics Control Law in Japan. However, various fly agaric-related products are now entering the Japanese market via the internet. In this study, fly agaric-related products available in this way were investigated for raw materials by DNA analysis and for additives by chemical analysis. Nucleotide sequence analysis of the mitochondrial 12S rDNA region suggested that these fly agaric-related products originate from A. muscaria or A. muscaria var. persicina. Furthermore, they were classified into three strains based on the ITS2-LSU nucleotide sequence. Harmine derivatives and/or tryptamine derivatives were detected in some of these products by LC/MS analysis. In accordance with this, the matK gene of Peganum harmala was found in all of the harmine derivative-containing samples.     

Introduction of sulfhydryl groups and disulfide linkage to mungbean 8Sα globulin and effects on physicochemical and functional properties

Sulfhydryl groups and disulfide bond were introduced in mungbean's major storage protein, 8Sα globulin, by protein engineering to improve structural stability and functional properties. Five modified proteins or mutants (F59C, I99C, A213C, one free sulfhydryl group; I99C/A213, one disulfide bridge; F59C/I99C/A213C, one free sulfhydryl group and one disulfide linkage) were expressed in Escherichia coli at a yield similar to that of the unmodified protein or wild type (WT) in soluble form (38%). The number of introduced groups in the mutants was confirmed by Ellman analysis. Mutant and WT proteins exhibited similar elution patterns on gel filtration indicating their trimeric native conformation. Mutants had 2 to 3.8 °C higher T_m values than WT and were digested by chymotrypsin at 52–58% in 60 min but exhibited different digestion patterns. All mutants showed greater hardness of heat-induced gels than WT, especially I99C/A213C and F59C/I99C/A213C. Results indicate the improved structural stability of the modified 8Sα globulin.     

New type of pressurized cultivation method providing oxygen for piezotolerant yeast

For efficient oxygen supply to pressurized culture, we developed a method using a highly pressurized membrane reactor with an air-saturated medium circulation system. The new method increased the cell growth of aerobic yeast approximately 20 folds larger than that in the case of using a conventional method.     

Plasma developable photoresist containing electronic excitation energy quenching system

A dry developable negative working resist composition comprised of poly(methyl isopropenyl ketone) (PMIPK) and 4-methyl-2,6-di(4′-azido-benzylidene) cyclohexanone-1 was examined. The main photochemical product formed in the resist pattern was found to be a secondary amine which crosslinks PMIPK. Post-annealing forms a hydrogen-bonded product which shows a powerful electronic excitation energy quenching effect. The quencher is more powerful than the aromatic compound arising from the azide by post-annealing only. The residual resist thickness of the negative pattern is about 80 percent of the initial thickness of the coating in spite of all the azide compound remaining in the resist coating. The obtained dry developed resist pattern has a high dry etch resistance. Etchings of Si and SiO₂ were performed by plasma and reactive ion etching, respectively.     

Electromechanical Properties of Porous Piezoelectric Ceramics

A theoretical approach is forwarded to predict the electromechanical properties of porous piezoelectric ceramics. The analysis is able to account for the effects of porosity shape and concentration and is applicable to piezoelectric ceramics of arbitrary material symmetry. By coupling the exact solution for a single ellipsoidal pore embedded in an infinite piezoelectric matrix with an effective medium approximation, the theory considers, in an approximate manner, interaction effects at finite porosity concentrations. The theoretical estimates are developed using a matrix formulation which enables all elastic, dielectric, and piezoelectric moduli of the porous solid to be readily computed. Numerical results are presented to illustrate the effects of the shape and concentration of the porosity on the effective electroelastic moduli and transducer parameters of practical importance. Particular attention is devoted to assessing the sensitivity of the effective electromechanical properties to the accuracy of the input data. Finally, theoretical estimates are shown to be in good agreement with existing experimental results for porous piezoelectric ceramics with various microstructural geometries.     

Influence of shearing history on the rheological properties and processability of branched polymers. III. An amorphous long-chain branched polymer

Viscoelastic properties of branched polymers vary with their shearing history; notwithstanding, their primary molecular parameters do not change. According to a recent study, such viscoelastic variation was believed to be observed only with crystalline long-chain branched polymers such as low-density polyethylene or polyacetal. That is, the origin of the viscoelastic variation was attributed to the presence of specific entanglements at the branching points, which was formed during the crystallization process. However, the viscoelastic variation of the long-chain branched polymers is the phenomenon at temperatures well above their melting points, namely, it is considered that whether the long-chain branched polymers are crystalline or not is not essential for the occurrence of the viscoelastic variation. Thus, the influence of the shearing history on the rheological properties of an amorphous long-chain branched polymer was investigated in this paper, and it was found that, irrespective of their crystallinity, the viscoelastic properties of long-chain branched polymers vary according to their shearing history.     

Properties of networks obtained by internal plasticization of epoxy resin with aromatic and aliphatic glycidyl compounds

In order to improve the fracture properties of p, p′-diaminodiphenylmethane-cured epoxy resin, various kinds of aromatic and aliphatic glycidyl compounds were investigated as a modifier at an amount of 30 wt %. Several compounds promoted the fracture toughness. In any glycidyl compounds, however, heat resistance was decreased by the modification. The dynamic mechanical properties of the modified epoxy resins were measured. The crosslinking density ρ was calculated from the theory of rubber elasticity, and the mechanical properties of the resins were discussed in regard to the crosslinking density. Tensile strength was scarcely affected by the crosslinking density. Elongation at break and Izod impact strength increased remarkably with decrease in crosslinking density. The fracture toughness K_Ic- increased with decrease in crosslinking density except at small ρ.     

Plasma polymerized membranes and gas permeability. II

Thin films were deposited onto porous substrates by plasma polymerization using three kinds of organosilicic compounds, tetramethylsilane (TMS), hexamethyldisiloxane (M₂), and octamethylcyclotetrasiloxane (D₄). Those composite membranes showed different characteristics of gas permeability. When D₄ was plasma-deposited onto a porous substrate, the composites membrane showed the highest oxygen permeability and the lowest oxygen-to-nitrogen permeability ratio. The composite membrane prepared from TMS showed the permeability characteristics opposite to the membrane obtained from D₄. Infrared spectrum of the polymer from D₄ resembles that of dimethylpolysiloxane. The plasma polymers from TMS and M₂ showed different profiles in Si? O absorption bands in the range 1100–1000 cm^-1 or in absorption bands of SiCH₃ groups in the range 850–750 cm^-1 from respective monomers. X-ray photoelectron spectroscopic observation indicated that all the plasma polymers contained more than two species of Si atom with different oxidation states. The greater part of Si atoms in plasma polymers took the same oxidation states in corresponding monomer. The gas permeability characteristics were closely related to the oxidation states of Si atom in the plasma polymers.     

Mechanism of Mechanical Strength Increase of Soda–Lime Glass by Aging

Two models have been proposed to explain the mechanical strength increase of abraded or indented soda–lime glasses upon aging, namely, crack tip blunting and the release of residual tensile stress near the crack tip. To clarify the mechanism, the time dependence of the strengthening of an abraded soda–lime glass was investigated. Effects of aging media, such as moist air, distilled water, 1 N HCI and 1 N NaOH solutions, as well as the abrasion flaw depth, were determined. The strength increase rate in water of abraded soda–lime glass was compared with those of borosilicate and high-silica glasses. The effect of stressing during aging was also investigated. It was found that the rate of strength increase was faster with decreasing abrasion flaw depth and with decreasing chemical durability. For a given flaw depth, an acidic solution produced the fastest strengthening. The strengthening rate was found to accelerate because of the "coaxing'effect of stressing during aging. From these observations, it was concluded that the strengthening rates relate to the diffusion process and chemical reactions, especially the alkali–hydrogen (or hydronium) ion-exchange reaction, near the crack tip. The role of the residual tensile stress appears to be similar to that of the applied tensile stress, helping the diffusion process near the crack tip. The observed strength increase of soda–lime glass by aging was thus attributed to the effective blunting of the crack tip geometry by the glass–water reaction.