Identification of Japanese Astraeus, based on morphological and phylogenetic analyses

To clarify the diversity of Astraeus in Japan, 35 collections of Astraeus basidiomata from Japan and Thailand were examined for their morphological characteristics and the nucleotide sequences of the rDNA ITS region and compared with several worldwide Astraeus species. Japanese Astraeus specimens were separated into two groups based on basidiome size, shape of exoperidium, and ornamentation and size of basidiospores. The phylogenetic tree analyses supported the separation, and the morphological groups belonged to different clades. The Japanese Astraeus group 1, morphologically matched to Astraeus hygromerticus var. koreanus, showed the closest phylogenetic relationship with Astraeus hygrometricus from North American and Mediterranean regions, suggesting that the Astraeus group 1 can be indentified as A. hygrometricus var. koreanus. Another Astraeus group, group 2, morphologically matched to A. hygrometricus s.l., showed a distinct monophyletic clade that was separated from A. hygrometricus complexes, indicating an undescribed species.     

Efficient amplification of light and heavy chain variable regions and construction of a non-immune phage scFv library

Non-immune phage scFv library is one of the most attractive resources for therapeutics, diagnostics and basic research. As a matter of fact, quality of the library is limited by inefficient PCR cloning of antibody genes using degenerated primers. PCR using this type of primers is difficult to optimize conditions for efficient amplification, and therefore causes loss of antibody diversities. To overcome this problem, we described a novel two-step amplification of V_κ and V_H genes with newly designed primer sets. Initially, we amplified V_κ and V_H genes from their signal sequences to the joining region to keep antibody diversity as large as possible. Thereafter, highly degenerated primers were used to amplify the V_κ and V_H genes from the framework region 1 to the joining region. The V_κ and V_H genes from the second PCR then were linked by PCR overlapping extension to generate the scFv library. Fifteen clones from the library were randomly picked and sequenced, and the diversity of full-length scFvs was confirmed. Expression capability of clones in the library was 80% after confirmation using colony hybridization. The results demonstrated the efficiency of this strategy and the primer sets for construction of the scFv library.     

Isolation of rhizospheric and roots endophytic actinomycetes from Leguminosae plant and their activities to inhibit soybean pathogen, Xanthomonas campestris pv. glycine

In this study, actinomycetes from roots and rhizospheric soils of leguminous plants were isolated using starch casein agar supplemented with antifungal and antibacterial antibiotics. Three hundred and seventeen actinomycetes were isolated with 77 isolates obtained from plant roots and 240 isolates from rhizospheric soils. Analysis of whole-organism hydrolysates showed that 289 strains were rich in the LL-isomer of diaminopimelic acid, a result consistent with their assignment to the streptomycetes. The remaining 28 strains were assigned to non-streptomycetes based on the presence of meso-isomer of diaminopimelic acid in cell wall. Sixty-four isolates (20.2 %) showed antagonistic activity against soybean pathogen Xanthomonas campestris pv. glycine by agar overlay method. Isolate RM 365 showed the highest activity with an inhibition ratio of 3.79, with no inhibitory activity on the growth of Rhizobium japonicum TISTR 079, Rhizobium sp. TISTR 061 and Rhizobium sp. TISTR 063. The 16S rRNA gene sequence analysis revealed that isolate RM 365 shared 99.28 % similarity to Streptomyces caeruleatus GIMN4^T (GQ329712). In addition, isolates which contained meso-DAP were also identified by 16S rRNA gene sequence analysis. The results showed that they were members of the genus Amycolatopsis, Isoptericola, Micromonospora, Microbispora, Nocardia, Nonomuraea, Promicromonospora and Pseudonocardia.     

Melitid amphipods from the Gulf of Thailand,with a description of Dulichiella pattaniensis,a new species

Two species of melitid amphipod were collected from the Gulf of Thailand. Dulichiella pattaniensis is new to science, and Melitalati latiflagella Ren & Andres, 2012 has not been previously reported from Thai Waters. Dulichiella pattaniensis is characterized by male gnathopod 2 distolateral crown with 4 spines; pleonite/urosomite formula 7-7-7-5-6-2; pereopod 5-7 dactylus with 2 accessory spines. This combination of characters has not been recorded previously in the Dulichiella. The characters of the specimens are described and illustrated. All specimens are deposited in the Princess Maha Chakri Sirindhorn Natural History Museum, Prince of Songkla University, Thailand.     

A DFT calculation on nonenzymatic degradation of isoaspartic residue

The mechanical properties of pristine and defective Si-based hybrid sheets are studied using molecular dynamics calculations for a temperature ranging from 100 to 800 K, in conjunction with a variable strain rate. When increasing temperature, the melting phase of the hybrids occurs from the solid to the liquid phase, while the increase in the strain rate enhances their elastic parameters. The absence of plastic stage reveals that the fracture pattern is brittle in these 2D materials. Under the uniaxial loading, the systems stretch, resulting in the failure of the crystalline skeletons that lose their rigidity with anisotropic behavior observed only for SiC. In defective hybrids, the point defects reduce the values of fracture strength and strain without affecting the brittle behavior of the sheets. The results impart that coupling high temperature to SiC material offers new possibilities for MEMS devices, whereas SiGe is a promising candidate for microelectronic devices.

     

The Awa1 gene is required for the foam-forming phenotype and cell surface hydrophobicity of sake yeast

Sake, a traditional alcoholic beverage in Japan, is brewed with sake yeasts, which are classified as Saccharomyces cerevisiae. Almost all sake yeasts form a thick foam layer on sake mash during the fermentation process because of their cell surface hydrophobicity, which increases the cells' affinity for bubbles. To reduce the amount of foam, nonfoaming mutants were bred from foaming sake yeasts. Nonfoaming mutants have hydrophilic cell surfaces and no affinity for bubbles. We have cloned a gene from a foam-forming sake yeast that confers foaming ability to a nonfoaming mutant. This gene was named AWA1 and structures of the gene and its product were analyzed. The N- and C-terminal regions of Awa1p have the characteristic sequences of a glycosylphosphatidylinositol anchor protein. The entire protein is rich in serine and threonine residues and has a lot of repetitive sequences. These results suggest that Awa1p is localized in the cell wall. This was confirmed by immunofluorescence microscopy and Western blotting analysis using hemagglutinin-tagged Awa1p. Moreover, an awa1 disruptant of sake yeast was hydrophilic and showed a nonfoaming phenotype in sake mash. We conclude that Awa1p is a cell wall protein and is required for the foam-forming phenotype and the cell surface hydrophobicity of sake yeast.     

Two new cycloartane-type triterpenoids and one new flavanone from the leaves of Dasymaschalon dasymaschalum and their biological activity

Two new cycloartane-type triterpenoids, 3β-hydroxy-21-O-acetyl-24-methylenecycloartane (3) and 3β,21-dihydroxy-24,31-epoxy-24-methylenecycloartane (4), one new flavanone, 7-hydroxy-6,8-dimethoxyflavanone (5), two new natural products, 2-hydroxybenzyl benzoate (7) and 2-phenyl-2-acetoxyethyl benzoate (8), and ten known compounds, 3β-hydroxy-24-methylenecycloartane (1), 3β,21-dihydroxy-24-methylenecycloartane (2), desmosdumotin B (6), artabotrene (9), (?)-senepoxide (10), (+)-crotepoxide (11), (?)-1,6-desoxypipoxide (12), rotundol (13), cassipourol (14) and (+)-spathulenol (15) were isolated from the leaves of Dasymaschalon dasymaschalum. The structures of the new compounds were elucidated by spectroscopic analysis and of the known compounds by comparison of their physical, UV, IR, ¹H and ¹³C NMR data with those of published compounds. Antimycobacterial, antiplasmodial and cytotoxic activities of the isolates, except 8 and 10 were evaluated. Compounds 1, 4, 5, 11, 12 and 15 exhibited potent cytotoxic activities against human lung cancer cell lines (NCI-H187) with respective IC₅₀ values of 4.67, 7.82, 1.85, 6.33, 3.07 and 6.68 μg/mL.     

Superior divisional oculomotor nerve palsy caused by midbrain neurocysticercosis

Superior divisional oculomotor nerve palsy caused by intrinsic brainstem disease occurs rarely. We herein report, to our knowledge, the first case of midbrain neurocysticercosis presenting as isolated superior divisional paresis of the oculomotor nerve. A 25-year-old woman presented with acute onset of headache and ptosis-upgaze palsy of the right eye. Results of the CT scan and MRI of the brain were compatible with neurocysticercosis at the left midbrain. She was dramatically responded to albendazole treatment. The diagnosis was confirmed by positive enzyme-linked immunosorbent assay (ELISA) test for cysticercus in her serum.