Four new Pythium species from aquatic environments in Japan.

Four new species of Pythium s.str., namely Pythium utonaiense, Pythium brachiatum, Pythium biforme and Pythium junctum, were described based on their morphology and molecular phylogeny. These species were isolated from freshwater samples collected from aquatic environments (pond and lake) in Japan. They were identified as belonging to the genus Pythium s.str. by their morphology, such as filamentous hyphal inflated structures, and molecular phylogenetic analyses based on the internal transcribed spacer region of the ribosomal RNA gene and mitochondrial cytochrome c oxidase subunit 1 gene. The new species are compared to closely related Pythium species regarding their phylogenetic positions and morphological features.

Nationwide multicenter study aimed at the establishment of common reference intervals for standardized clinical laboratory tests in Japan.

BACKGROUND: The Japanese Association of Medical Technologists (JAMT) sought to establish common reference intervals (RIs) applicable nationwide in Japan for 27 serum constituent analytes for which certified reference materials are available and nine analytes frequently measured in routine tests. More than 100 laboratories certified for metrological traceability collaborated in the recruitment, sampling, and measurement of analytes for the establishment of RIs. No previous attempt has been made to establish RIs by such a large number of laboratories. The allowable limits of trueness and intermediate precision based on the JAMT criteria were applied to the reference values measured by these laboratories, and measured values within the allowance limits were used to establish RIs. METHODS: Reference individuals included 5748 healthy volunteers aged 18-65 years who were engaged in medical care-related work based on the CLSI guidelines. After secondary exclusion of individuals in whom abnormal values were detected in basic routine test items and adjustment for the distribution of age and gender, 3371 reference individuals were chosen in the parametric determination of RIs. Employing the three-level nested ANOVA, between-laboratory, -region, -sex, and -age variations were evaluated. RESULTS: No significant difference was noted in between-region variations in any item. Results of ANOVA revealed between-sex and -age variations in 14 and 15 analytes, respectively. Based on these results of variation, RIs were established with and without partition by sex. CONCLUSIONS: Since no between-region variation was detected in reference values among accuracy-certified core laboratories, RIs applicable nationwide were established.

Molecular phylogenetic analyses based on the nuclear rRNA genes and the intron-exon structures of the nuSSU rRNA gene in Dictyocatenulata alba (anamorphic Ascomycota).

Molecular phylogenies inferred from the nuclear small subunit rRNA gene (nuSSU), nuclear large subunit rRNA gene D1/D2 region (nuLSU), and ITS-5.8S rRNA gene (ITS) indicated that five cultures of the lichenized hyphomycete Dictyocatenulata alba isolated from Japan form a monophyletic clade with high bootstrap support, and a close relationship to the Ostropomycetidae (Lecanoromycetes, Pezizomycotina, Ascomycota). Insertion sequences were found in the nuSSU of all isolates [e.g., nine insertions in the strain JCM 5358 (Japan Collection of Microorganisms)], some of which were group I introns. Five new insertion positions were found among the D. alba isolates. Using BLAST, none of the insertion sequences of D. alba were closely related to those of fungi or other organisms deposited in public DNA databases. Insertion positions were similar in some isolates, and two positions were common to all isolates. Although all phylogenetic analyses based on nuSSU, nuLSU, and ITS revealed the monophyly of D. alba, the isolates were divided into two (in the nuSSU tree) or three (in the nuLSU and ITS trees) groups. Based on the phylogenetic analyses and the intron-exon structures, the five isolates identified as D. alba belong to three cryptic species and therefore D. alba should be considered a species complex. The very slow-growing, tough agar colonies of the isolates, the occurrence of the species on both slightly lichenized and nonlichenized surfaces of trees, or pebbles (rarely on soil), suggest that the members of the D. alba complex may be lichenized. The photobiont was not clearly identified in this study.

Fungus symbionts colonizing the galleries of the ambrosia beetle Platypus quercivorus.

Isolations were made to determine the fungal symbionts colonizing Platypus quercivorus beetle galleries of dead or dying Quercus laurifolia, Castanopsis cuspidata, Quercus serrata, Quercus crispula, and Quercus robur. For these studies, logs from oak wilt-killed trees were collected from Kyoto Prefecture, Japan. Fungi were isolated from the: (1) entrances of beetle galleries, (2) vertical galleries, (3) lateral galleries, and (4) the larval cradle of P. quercivorus in each host tree. Among the fungus colonies which appeared on YM agar plates, 1,219 were isolated as the representative isolates for fungus species inhabiting in the galleries based on their cultural characteristics. The validity of the visual classification of the fungus colonies was checked and if necessary properly corrected using microsatellite-primed PCR fingerprints. The nucleotide sequence of the D1/D2 region of the large subunit nuclear rRNA gene detected 38 fungus species (104 strains) of which three species, i.e., Candida sp. 3, Candida kashinagacola (both yeasts), and the filamentous fungus Raffaelea quercivora were isolated from all the tree species. The two yeasts were most prevalent in the interior of galleries, regardless of host tree species, suggesting their close association with the beetle. A culture-independent method, terminal restriction fragment length polymorphism (T-RFLP) analysis was also used to characterize the fungus flora of beetle galleries. T-RFLP patterns showed that yeast species belonging to the genus Ambrosiozyma frequently occurred on the gallery walls along with the two Candida species. Ours is the first report showing the specific fungi inhabiting the galleries of a platypodid ambrosia beetle.

RK-95113, a new angiogenesis inhibitor produced by Aspergillus fumigatus.

We found that the strain Aspergillus fumigatus var. fumigatus RK95-113 produced a new angiogenesis inhibitor, RK-95113. Spectroscopic analyses identified the structure of RK-95113 as a new fumagillin-related small molecule. RK-95113 preferentially inhibited the growth of human umbilical vein endothelial cells (HUVECs) rather than that of human normal fibroblasts in cell proliferation assays and blocked endothlial cell migration induced by vascular endothelial growth factor (VEGF).