Diversity of 16S ribosomal DNA-defined bacterial population in acid rock drainage from Japanese pyrite mine

Four acidophilic bacteria (YARDs1-4) were isolated from an acid rock drainage (ARD) from Yanahara mine, Okayama prefecture, Japan. The physiological and 16S rDNA sequence analyses revealed that YARD1 was closely affiliated with Acidithiobacillus ferrooxidans, YARD2 was an Acidiphilium-like bacterium, and YARD3 and YARD4 were sulfur-oxidizing bacteria with a relatively close relationship to A. ferrooxidans in the phylogenetic analysis. A molecular approach based on the construction of a 16S rDNA clone library was used to investigate the microbial population of the ARD. Small-subunit rRNA genes were PCR amplified, subsequently cloned and screened for variation by a restriction fragment length polymorphism (RFLP) analysis. A total of 284 clones were grouped into 133 operational taxonomic units (OTUs) by the RFLP analysis. Among them, an OTU showing the same RFLP pattern as those of the isolates from the ARD was not detected. The phylogenetic analysis based on the 16S rDNA sequences from 10 major OTUs and their close relatives revealed that 4 OTUs containing 32.1% of the total clones were loosely affiliated with Verrucomicrobia, 2 OTUs containing 6.6% of the total clones were loosely affiliated with Chloribi, and other OTUs were affiliated with Actinobacteria, Nitrospirae, and beta-Proteobacteria.     

Differential reprogramming of somatic cell nuclei after transfer into mouse cleavage stage blastomeres

Mammalian somatic cell cloning requires factors specific to the oocyte for reprogramming to succeed. This does not exclude that reprogramming continues during the zygote and cleavage stages. The capacity or role of zygotic and cleavage stages to reprogram somatic cell nuclei is difficult to assess due to the limited development of somatic cell nuclei transplanted into cytoplasts of these stages. Alternatively, tetraploid embryos have been used to study reprogramming and can be assessed for their contribution to extra-embryonic lineages. When mouse cumulus cell nuclei transgenic for Oct4-green fluorescent protein (GFP) were injected into intact two- and four-cell stage blastomeres, manipulated embryos developed into blastocysts with expression of Oct4-GFP as observed in embryos produced by nuclear transfer into metaphase II oocytes. However, only the latter contributed to extra-embryonic tissues in day 10.5 conceptuses, with the exclusion of the somatic genome in cells originating from transfer into blastomeres already at 5.5 days post conception. Somatic nuclei transferred into cleavage stage blastomeres reinitiated expression of an embyronic-specific transgene, but lacked the extent of reprogramming required for contribution to postimplantation development, even when complemented by an embryonic genome.     

Mating Communication Systems of Four Plusiinae Species Distributed in Japan: Identification of the Sex Pheromones and Field Evaluation

The sex pheromones of four Plusiinae species in the family Noctuidae, Ctenoplusia albostriata (CA), Macdunnoughia purissima (MP), Syngrapha ain (SA), and Diachrysia stenochrysis (DS) were identified by GC-EAD and GC-MS analyses. These were subsequently evaluated by a series of field trials in different ecological habitats. CA females produced (Z)-5-decenyl acetate (I), (Z)-7-dodecenyl acetate (II), and (Z)-7-dodecen-1-ol (III) in a ratio of 2:100:13. While the antennae of CA males responded to all three components, neither I nor III was essential to capture males. The MP females produced II, III, and (Z)-5-dodecenyl acetate (IV) in a ratio of 100:80:20, and this blend captured the most MP males. Compounds II and III were also identified from SA females, and mixtures ranging from 4:1 to 2:3 were equally attractive to males. DS females only produced one active compound, (Z)-7-decenyl acetate (V), and DS is the first Plusiinae species identified as using only the ω3-compound and none of ω5-compounds, such as II and III, which are common components of Plusiinae pheromones. Using previously published chemical and biosynthetic data, together with the finding from this study, we propose a classification of the Plusiinae sex pheromones.     

Formation of refractory ring-expanded triazine intermediates during the photocatalyzed mineralization of the endocrine disruptor amitrole and related triazole derivatives at UV-irradiated TiO2/H2O interfaces

Amitrole (ATz, 3-amino-1H-1,2,4-triazole) is a widely employed herbicide with strong estrogenic activity that can lead to abnormalities of the thyroid gland and can cause mutations. The photocatalytic transformation of ATz was carried out at the UV-irradiated TiO2/H2O interface, along with the triazole derivatives Tz (1H-1,2,4-triazole) and DaTz (3,5-diamino-1H-1,2,4-triazole) to assess the decomposition of these herbicides, to identify intermediates, and to elucidate some mechanistic details of the ATz degradation. Conversion of the nitrogens of these triazoles to NH4+ and/ or NO3- ions occurs competitively and depends on the number of amine functions on the five-membered triazole rings. Photomineralization of the substrates in terms of loss of nitrogen to NH4+/NO3- was rather low (ca. 25-40%) for each of the triazoles, whereas evolution of CO2 (loss of TOC) was more significant (60-70%), indicating considerable retention of nitrogen in the intermediate products. UV-Vis spectroscopy, TOC assays, FT-IR spectroscopy, proton NMR spectrometry, electrospray LC-MS, and molecular orbital calculations were brought to bear in assessing the temporal course of the photocatalyzed process(es). Results show that after cleavage of the triazole ring, the various intermediate fragments recombine to yield ring-expanded six-membered triazine intermediates, which slowly degrade to give the refractory cyanuric acid under the conditions used.     

Malonaldehyde content in some breads and its change during storage

Malonaldehyde (MA) content in plain bread, French bread, and croissants was analyzed. MA was detected in all the analyzed samples. The MA and lipid contents in croissants were higher than those in other breads. Changes in MA content in plain bread, French bread, and croissants stored at 4 degrees C were examined after 0, 2, and 4 days of storage. The MA content in all the bread samples increased during the storage period. The highest increase of MA content was observed in French bread.     

Permeation, diffusion and dissolution of hydrogen isotopes, methane and inert gases through/in a tetrafluoroethylene film

Tetrafluoroethylene (TFE) is widely used for conventional tritium handling systems such as vacuum seals, tubing and so on. We measured the permeation of the three hydrogen isotopes, methane and the inert gases through a TFE film at room temperature by means of the time-lag method in order to establish the physicochemical properties which determine the solubility and diffusivity of those gases. It was found that the diffusion constant of the inert gases changed exponentially with the heat of vaporization and the solubility was an exponential function of the Lennard-Jones force constant of the gases. On the other hand, hydrogen isotopes and methane deviated from these relations. It is concluded that chemical interactions between the solute and the solvent play an important role for the dissolution and the diffusion of these gases in TFE.     

Variation in response to dexamethasone of a patient with Cushing's syndrome

A patient with Cushing's syndrome due to a nonresectable chromophobe adenoma underwent external irradiation of the hypothalamic-pituitary area. The signs of Cushing's syndrome ameliorated subsequently but recurred eventually, necessitating bilateral adrenalectomy 30 months after initial presentation. Response to dexamethasone suppression testing on four occasions varied greatly. These results may reflect alterations in sensitivity of the tumour or the hypothalamic-pituitary axis to feedback inhibition by glucocorticoids.