From Terpenoids to Aliphatic Acids: Further Evidence for Late-Instar Switch in Osmeterial Defense as a Characteristic Trait of Swallowtail Butterflies in the Tribe Papilionini

We compared the chemical compositions of the osmeterial secretions of fourth and fifth (last) instars of eight swallowtail species of the tribe Papilionini. Four species (Papilio demoleus, P. polytes, P. paris, and P. macilentus) are Asian Rutaceae-feeding swallowtails. The other four (Chilasa epicydes, C. agestor, P. troilus, and P. glaucus) represent more distant clades within the Papilionini and species with larval hosts in other plant families. We conducted a quantitative analysis for six species, but only qualitative analysis for P. glaucus and C. agestor. In all eight species, regardless of larval host plant, secretions of the fourth instar principally consisted of mono- and sesquiterpene hydrocarbons, whereas those of the fifth instar comprised aliphatic acids and their esters. Consistent with earlier findings, our results suggest that this “heterogeneous” pattern of osmeterial chemistry, not seen in other tribes, may characterize the Papilionini as a whole. Unlike those of most Papilio species, the fourth and fifth instars of Chilasa species resemble each other in body coloration. Thus, the heterogeneous osmeterial pattern is not necessarily associated with color change in papilionid larvae. The major terpenoids identified in fourth instar larval secretions from the six species were α-pinene, sabinene, β-myrcene, limonene, β-phellandrene, (Z)-β-ocimene, (E)-β-ocimene, p-mentha-1,4(8)-diene, β-elemene, β-caryophyllene, (E)-β-farnesene, (3Z,6E)-α-farnesene, (Z)-α-bisabolene, germacrene-A, (E)-α-bisabolene, and germacrene-B. The profiles for individual species differed both qualitatively and quantitatively from one another, and certain species also secreted methyl 3-hydroxy-n-butyrate and oxygenated sesquiterpenes in relatively large proportions. Secretions from fifth instars were composed of varying proportions of isobutyric, 2-methylbutyric, and acetic acids, and methyl and ethyl (minor) esters of both isobutyric and 2-methylbutyric acids. The heterogeneity of osmeterial chemistry in the tribe Papilionini may represent fine-tuning of chemical defense in response to shifting predation pressures as the larvae age and grow.     

PCR-mediated seamless gene deletion and marker recycling in Saccharomyces cerevisiae

Repeated gene manipulations can be performed in yeast by excision of an introduced marker. Cassette modules containing a marker flanked by two direct repeat sequences of hisG or loxP have often been used for marker recycling, but these leave one copy of the repeats in the chromosome after excision. Genomic copies of a repeat can cause increased mistargeting of constructs containing the same repeats or unexpected chromosomal rearrangements via intra- or interchromosomal recombinations. Here, we describe a novel marker recycling procedure that leaves no scar in the genome, which we have designated seamless gene deletion. A 40 base sequence derived from an adjacent region to the targeted locus was placed in an integrating construct to generate direct repeats after integration. Seamless HIS3 deletion was achieved via a PCR fragment that consisted of a URA3 marker attached to a 40 base repeat-generating sequence flanked by HIS3 targeting sequences at both ends. Transformation of the designed construct resulted in his3 disruption and the generation of 40 base direct repeats on both sides of URA3 in the targeted locus. The resulting his3::URA3 disruptants were plated on 5-fluoroorotic acid medium to select for URA3 loss. All the selected colonies had lost URA3 precisely by recombination between the repeats, resulting in his3 deletion without any extraneous sequences left behind in the chromosome.     

Thermochemical Nonequilibrium 2D Modeling of Nitrogen Inductively Coupled Plasma Flow

Two-dimensional (2D) numerical simulations of thermochemical nonequilibrium in?ductively coupled plasma (ICP) flows inside a 10-kW inductively coupled plasma wind tunnel (ICPWT) were carried out with nitrogen as the working gas. Compressible axisymmetric Navier-Stokes (N-S) equations coupled with magnetic vector potential equations were solved. A four-temperature model including an improved electron-vibration relaxation time was used to model the internal energy exchange between electron and heavy particles. The third-order accuracy elec?tron transport properties (3rd AETP) were applied to the simulations. A hybrid chemical kinetic model was adopted to model the chemical nonequilibrium process. The flow characteristics such as thermal nonequilibrium, inductive discharge, e?ects of Lorentz force were made clear through the present study. It was clarified that the thermal nonequilibrium model played an important role in properly predicting the temperature field. The prediction accuracy can be improved by applying the 3rd AETP to the simulation for this ICPWT.     

Production of cellulose- and xylan-degrading enzymes by a koji mold, aspergillus oryzae, and their contribution to the maceration of rice endosperm cell wall

The production of cellulose- (CEL), xylan- (XYL), and pectin-degrading enzymes (PEC) by a koji mold, Aspergillus oryzae, was studied, and their contributions to the maceration of the rice endosperm cell wall were investigated with regard to the utilization of available rice in the sake mash. The sake koji mold showed higher CEL and XYL productivities, whereas the miso and soy sauce koji molds showed higher PEC productivity. Statistical analyses indicated that CEL and XYL contribute predominantly and synergistically to the maceration of the rice endosperm cell wall. A. oryzae produced at least three kinds of CEL (Cel-1, 2, 3) and two kinds of XYL (Xyl-1, 2) when cultured in a wheat bran medium. In the solid-state culture, the production of Cel-3 and Xyl-2 was markedly stimulated by decreasing the moisture content of the solid substrate, although the production levels of Cel-1 and Xyl-1 were almost the same. These data suggest that the production of Cel-3 and Xyl-2 is strongly influenced by culture conditions, and that water activity is one of the dominant factors in the regulation of their production.     

Rhabdomyolysis after infection and taking a cold medicine in a patient who was susceptible to malignant hyperthermia

OBJECTIVE: To investigate the role of human parvovirus B19 (Parvo B19), cytomegalovirus (CMV) and human papilloma virus (HPV) viruses in the aetiopathogenesis of spontaneous abortions. STUDY DESIGN: Abortion material from 102 cases of women with spontaneous abortions were analysed for the presence of Parvo B19, CMV and HPV DNA using the polymerase chain reaction (PCR) technique. Serological assays were used for the detection of specific IgM and IgG antibodies against Parvo B19 virus and CMV in the maternal sera. RESULTS: Parvo B19 virus genome was detected in two cases of spontaneous abortion, by PCR amplification, while CMV and HPV genomes were not observed. Serological markers were indicative for Parvo B19 virus and CMV infection in ten and four cases, respectively. CONCLUSIONS: PCR is a useful method for investigating the viral contribution to the aetiopathogenesis of spontaneous abortions and for detecting the viral genome in the abortion material. This study of 102 cases of spontaneous abortion does not implicate CMV and HPV in the aetiopathogenesis of spontaneous abortion, although it indicates a possible abortional role for Parvo B19 virus.     

Material design for polymer/azobenzene liquid crystal/metal salt composite films possessing thermo- and photoresponsive ionic conductivity

Various azobenzene derivatives and polymers were attempted as materials for thin composite films containing a polymer, a liquid crystal, and lithium perchlorate which can undergo ionic-conductivity switching induced by light or heat. Poly(vinyl chloride) -based composite films containing enantiotropic azobenzene liquid crystals 1 , exhibited significant ionicconductivity change based on thermo- and photoinduced phase transitions of 1 , the temperature dependence on the ionic conductivity reflecting the enantiotropic phase transition behavior of the liquid crystals. Marked, reversible ionic-conductivity switching on alternating irradiation of UV and visible lights was attained with the composite films containing 1 , especially 1 (n = 12). Incorporation of monotropic liquid crystals 3 to the composite films allowed bistability in the temperature dependence on the ionic conductivity. Employment of a polycarbonate resin and poly(methyl methacrylate) as the polymer material of the composite films gave similar ionic-conductivity changes to the poly(vinyl chloride) composite films, whereas polyethyleneoxide brought about some different results in the photoinduced ionic-conductivity change.     

Cation Deficiency in Bi2(Sr,Ca)3Cu2O8+d

X-ray lattice parameter measurements of the Bi_{2- x} (Sr,Ca)_{3- y} Cu_{2- z} O_{8+ d} system show that a continuously varying solid-solution cation nonstoichiometry exists within the overall composition ranges x < 0.32, -0.2 < y < 0.32, and z < 0.25. A modulated structure along the b axis remains present over these composition ranges. Defects introduced by cation deficiency have been clarified to be vacancies by density measurements. The oxygen content determined by titration decreases proportionately with increasing cation vacancy concentrations. In Bi-deficient compositions, the interstitial oxygen ion concentration decreased and the oxygen vacancy concentration increased with increasing concentration of Bi vacancies. The critical temperature, T _c , shows a maximum value at the stoichiometric cation composition and decreases to a slightly lower (5 to 10 K) constant value across the nonstoichiometric composition region with both increasing alkaline-earth and Cu deficiency.     

Differential Utilization of Pyrrolizidine Alkaloids by Males of a Danaid Butterfly, <Emphasis Type="Italic">Parantica sita</Emphasis>, for the Production of Danaidone in the Alar Scent Organ

Males of the chestnut tiger butterfly, Parantica sita, secrete danaidone as a major component from the alar androconial organ (sex brand). Since danaidone has been postulated to be derived from various pyrrolizidine alkaloids (PAs), which males ingest as adults from PA-containing plants, we conducted oral administration tests of several PAs to examine their availability for danaidone production by P. sita males. Males fed with a mixture of intermedine (80%) and lycopsamine (20%) produced danaidone at an average of 25.7 g per individual, which was comparable to that found in field-caught males. In contrast, a smaller amount of danaidone (5.7 to 7.0 g/male) was formed when males ingested retronecine or heliotrine, and those fed with an HCl salt of monocrotaline or retrorsine produced only traces of danaidone (< 0.5 g/male). In addition, males showed a strong feeding response to intermedine/lycopsamine, whereas the other PAs elicited no positive feeding behavior. These results indicate that, unlike the arctiid moths, P. sita males can only successfully convert limited chemical types of PAs into danaidone, and further suggest that in the field, males selectively ingest particular PAs that are readily transformable into danaidone.     

Electrochemical epitaxial polymerization of single-molecular wires

The fabrication of organized molecular-scale structures is key to realizing high-performance molecular- and conjugated-polymer devices. Here, we demonstrate a unique single-molecular processing technique using electrochemistry, termed 'electrochemical epitaxial polymerization'. This technique is based on step-by-step electropolymerization of the monomer by applying voltage pulses to a monomer-electrolyte solution that also contains iodine. Using this technique, we observe the formation of high-density arrays of single conjugated-polymer wires as long as 75 nm on the surface of a Au(111) electrode. Our findings unveil the mechanism of electropolymerization, showing that the conjugated polymer wires grow from nuclei adsorbed on the iodine-covered Au(111) surface. The results may also open the door to mass-production of molecular-scale devices based on conjugated polymers.