Amanita satotamagotake sp. nov., a cryptic species formerly included in Amanita caesareoides.

We evaluated the inclusion of a cryptic species in a Japanese Amanita caesareoides population. We sampled A. caesareoides specimens under various vegetation and climate conditions, and then conducted phylogenetic analyses on sequences from seven loci. The A. caesareoides specimens showed two distinct groups, except when the ITS phylogeny was considered. These two phylogroups showed different distributions: subalpine-cool temperate and temperate-subtropical areas. Although these two phylogroups overlapped in terms of basidiospore size, the latter tended to exhibit smaller basidiospores. In addition, only the former showed mycelial growth on nutrient agar. Based on these phylo-morpho-ecophysiological characteristics, we separated the specimens labeled with the name A. caesareoides into two species. As the lectotype of A. caesareoides showed similarity to the former by DNA analysis, the latter was described as a new species, namely A. satotamagotake. Based on the geographic patterns of the two species, A. satotamagotake may have invaded the natural habit of A. caesareoides because of global warming.

PEG10 counteracts signaling pathways of TGF-ß and BMP to regulate growth, motility and invasion of SW1353 chondrosarcoma cells.

Recently, we reported highly active transforming growth factor (TGF)-ß and bone morphogenetic protein (BMP) signaling in human chondrosarcoma samples and concurrent downregulation of paternally expressed gene 10 (PEG10). PEG10 expression was suppressed by TGF-ß signaling, and PEG10 interfered with the TGF-ß and BMP-SMAD pathways in chondrosarcoma cells. However, the roles of PEG10 in bone tumors, including chondrosarcoma, remain unknown. Here, we report that PEG10 promotes SW1353 chondrosarcoma cell growth by preventing TGF-ß1-mediated suppression. In contrast, PEG10 knockdown augments the TGF-ß1-induced motility of SW1353 cells. Individually, TGF-ß1 and PEG10 siRNA increase AKT phosphorylation, whereas an AKT inhibitor, MK2206, mitigates the effect of PEG10 silencing on cell migration. SW1353 cell invasion was enhanced by BMP-6, which was further increased by PEG10 silencing. The effect of siPEG10 was suppressed by inhibitors of matrix metalloproteinase (MMP). BMP-6 induced expression of MMP-1, -3, and -13, and PEG10 lentivirus or PEG10 siRNA downregulated or further upregulated these MMPs, respectively. PEG10 siRNA increased BMP-6-induced phosphorylation of p38 MAPK and AKT, whereas the p38 inhibitor SB203580 and MK2206 diminished SW1353 cell invasion by PEG10 siRNA. SB203580 and MK2206 impeded the enhancing effect of PEG10 siRNA on the BMP-6-induced expression of MMP-1, -3, and -13. Our findings suggest dual functions for PEG10: accelerating cell growth by suppressing TGF-ß signaling and inhibiting cell motility and invasion by interfering with TGF-ß and BMP signaling via the AKT and p38 pathways, respectively. Thus, PEG10 might be a molecular target for suppressing the aggressive phenotypes of chondrosarcoma cells.

Removal of a migrated acupuncture needle from the cervical spinal canal with removal confirmation by cone-beam computed tomography in a hybrid operating room.

Background: Acupuncture has become one of the most popular alternative medical treatments in the world. However, if the needle is inserted incorrectly into the body, various adverse events can occur and, in such cases, the needle should be removed. Acupuncture needles are very thin and fragile, making them difficult to detect and to confirm breakage or residual needle fragments during surgery. We report a case of a patient's self-placed acupuncture needle migrating into the cervical spinal canal and its surgical removal. We used cone-beam computed tomography in the hybrid operating room to confirm that the needle was removed in its entirety. Case presentation: A 37-year-old man presented with neck pain and gait disturbance.While he was self-acupuncturing, an acupuncture needle accidentally broke, and the remaining part of the needle penetrated and made contact with the cervical spinal cord. Cervical spine radiographs showed a metallic foreign body between the C1 and C2 spinous processes in the direction of the anterior cervical spine. Computed tomography images revealed that the acupuncture needle was penetrating the spinal canal and was in contact with the cervical cord. The acupuncture needle was removed under general anesthesia. The use of cone-beam computed tomography in the hybrid operating room allowed intraoperative confirmation that there was no breakage during needle removal, and no needle fragments were left behind. His symptoms disappeared without any complications after the operation. Conclusion: To the best of our knowledge, this is the first report of the removal of an acupuncture needle that migrated into the cervical spinal canal using cone-beam computed tomography in a hybrid operating room. Intraoperative cone-beam computed tomography is useful in patients with small, fragile foreign bodies for confirmation of the location of the object and to check for the presence of residual fragments.

PCR-Based Method for the Detection of Toxic Mushrooms Causing Food-Poisoning Incidents.

In this study, species-specific identification of five toxic mushrooms, Chlorophyllum molybdites, Gymnopilus junonius, Hypholoma fasciculare, Pleurocybella porrigens, and Tricholoma ustale, which have been involved in food-poisoning incidents in Japan, was investigated. Specific primer pairs targeting internal transcribed spacer (ITS) regions were designed for PCR detection. The specific amplicons were obtained from fresh, cooked, and simulated gastric fluid (SGF)-treated samples. No amplicons were detected from other mushrooms with similar morphology. Our method using one-step extraction of mushrooms allows rapid detection within 2.5 hr. It could be utilized for rapid identification or screening of toxic mushrooms.

Osteoarthritis of the hip joint in elderly patients is most commonly atrophic, with low parameters of acetabular dysplasia and possible involvement of osteoporosis.

As elderly patients with hip osteoarthritis aged, acetabular dysplasia parameters decreased (Sharp's angle, acetabular roof obliquity angle, and acetabular head index) and the incidence of the atrophic type increased. Vertebral body fracture was more frequent in the atrophic type, suggesting the involvement of osteoporosis at the onset of hip osteoarthritis. INTRODUCTION: Osteoarthritis (OA) is associated with increased bone formation at a local site. However, excessive bone resorption has also been found to occur in the early stages of OA. Osteoporosis may be involved in the onset of OA in elderly patients. We conducted a cross-sectional radiographic study of patients with hip OA and examined the association between age and factors of acetabular dysplasia (Sharp's angle, acetabular roof obliquity angle, and acetabular head index) as well as the osteoblastic response to determine the potential involvement of osteoporosis. METHODS: This study included 366 patients (58 men, 308 women) who had undergone total hip arthroplasty for the diagnosis of hip OA. We measured the parameters of acetabular dysplasia using preoperative frontal X-ray images and evaluated each patient according to Bombelli classification of OA (hypertrophic, normotrophic, or atrophic type). RESULTS: As the patients aged, the parameters of acetabular dysplasia decreased. The incidence of the atrophic type of OA was significantly higher in older patients. Vertebral body fractures were more frequent in the atrophic type than in the other types. Additionally, the index of acetabular dysplasia was lower in the atrophic type. By contrast, the hypertrophic type was present in relatively younger patients and was associated with an increased index of acetabular dysplasia. CONCLUSION: In elderly patients with hip OA, the parameters of acetabular dysplasia decreased and the incidence of the atrophic type increased as the patients aged. The frequency of vertebral body fracture was high in patients with the atrophic type, suggesting the involvement of osteoporosis in the onset of hip OA.

TGF-ß signalling and PEG10 are mutually exclusive and inhibitory in chondrosarcoma cells.

Histological distinction between enchondroma and chondrosarcoma is difficult because of a lack of definitive biomarkers. Here, we found highly active transforming growth factor-ß (TGF-ß) and bone morphogenetic protein (BMP) signalling in human chondrosarcomas compared with enchondromas by immunohistochemistry of phosphorylated SMAD3 and SMAD1/5. In contrast, the chondrogenic master regulator SOX9 was dramatically down-regulated in grade 1 chondrosarcoma. Paternally expressed gene 10 (PEG10) was identified by microarray analysis as a gene overexpressed in chondrosarcoma SW1353 and Hs 819.T cells compared with C28/I2 normal chondrocytes, while TGF-ß1 treatment, mimicking higher grade tumour conditions, suppressed PEG10 expression. Enchondroma samples exhibited stronger expression of PEG10 compared with chondrosarcomas, suggesting a negative association of PEG10 with malignant cartilage tumours. In chondrosarcoma cell lines, application of the TGF-ß signalling inhibitor, SB431542, increased the protein level of PEG10. Reporter assays revealed that PEG10 repressed TGF-ß and BMP signalling, which are both SMAD pathways, whereas PEG10 knockdown increased the level of phosphorylated SMAD3 and SMAD1/5/9. Our results indicate that mutually exclusive expression of PEG10 and phosphorylated SMADs in combination with differentially expressed SOX9 is an index to distinguish between enchondroma and chondrosarcoma, while PEG10 and TGF-ß signalling are mutually inhibitory in chondrosarcoma cells.

Discrimination of mushrooms causing food-poisoning incidents by using DNA sequence analysis.

In this study, the identification of mushrooms by using DNA analysis was investigated. Our analysis of internal transcribed spacer (ITS) regions revealed that a DNA-based method could be applicable for samples that are difficult to distinguish in terms of the morphological characteristics. PCR amplification using templates extracted from cooked samples gave sufficient fragments to analyze the sequence. However, treatment with simulated gastric fluid (SGF) for more than 30 min affected the analysis of the ITS region. Application to samples of vomit is also discussed.

Size-selective diffusion in nanoporous but flexible membranes for glucose sensors.

A series of nanoporous membranes prepared from polyethylene-block-polystyrene were applied for size-selective diffusion of glucose and albumin molecules. Millimeter-sized test cells for characterization of such molecular diffusions were designed assuming an implantable glucose sensor. The prepared nanoporous membrane exhibits excellent flexibility and toughness compared to conventional nanoporous membranes of brittle alumina. Pore size of the membranes could be controlled from 5 to 30 nm by varying preparation conditions. All of these nanoporous membranes prepared in this study let glucose pass through, indicating a continuous pore connection through the entire thickness of the membrane in a few tens of micrometers. In contrast, membranes prepared under optimum conditions could perfectly block albumin permeation. This means that these vital molecules having different sizes can be selectively diffused through the nanoporous membranes. Such a successful combination of size selectivity of molecular diffusion in nanoscale and superior mechanical properties in macroscale is also beneficial for other devices requesting down-sized manufacture.

Zinc- and indium-mediated ring-expansion reaction of alpha-halomethyl cyclic beta-keto esters in aqueous alcohol.

Radical ring-expansion reaction of various alpha-halomethyl cyclic beta-keto esters and alpha-halomethyl benzocyclic beta-keto esters and chain-extension reaction of alpha-halomethyl beta-keto esters with zinc powder and indium powder in refluxing aqueous alcohol were carried out to generate the corresponding ring-expansion and chain-extension products. As the results indicate, it was found zinc powder was more effective than indium powder to give the corresponding ring-expanded products in quite good yields. Moreover, the addition of zinc bromide as Lewis acid showed an increase of the yields, depending on the substrates. The present reaction conditions are highly effective, simple, and environmentally friendly.