Mutations in the fused in sarcoma (FUS, also known as translated in liposarcoma) gene have been recently discovered to be associated with familial amyotrophic lateral sclerosis (FALS) in African, European and American populations. In a Japanese family with FALS, we found the R521C FUS mutation, which has been reported to be found in various ethnic backgrounds. The family history revealed 23 patients with FALS among 46 family members, suggesting a 100% penetrance rate. They developed muscle weakness at an average age of 35.3 years, followed by dysarthria, dysphagia, spasticity and muscle atrophy. The average age of death was 37.2 years. Neuropathological examination of the index case revealed remarkable atrophy of the brainstem tegmentum characterized by cytoplasmic basophilic inclusion bodies in the neurons of the brainstem. We screened 40 FALS families in Japan and found 4 mutations (S513P, K510E, R514S, H517P) in exon 14 and 15 of FUS. Even in Asian races, FALS with FUS mutations may have the common characteristics of early onset, rapid progress and high penetrance rate, although in patients with the S513P mutation it was late-onset. Degeneration in multiple systems and cytoplasmic basophilic inclusion bodies were found in the autopsied cases.Journal of Human Genetics advance online publication, 12 March 2010; doi:10.1038/jhg.2010.16.

We report two Creutzfeldt-Jakob disease (CJD) patients with rhythmic pupillary and palpebral oscillation who had a mutation of prion protein codon 200 that resulted in the substitution of lysine for glutamate (Glu/Lys). Alternating dilation and constriction of the pupils combined with elevation and descent of the eyelids occurred in correspondence with periodic sharp wave complexes (PSWCs) on the electroencephalogram and with myoclonus of the head, face, and extremities. The onset of pupillary dilation and palpebral elevation coincided with the PSWCs. Initiation of these rhythmic pupillary and palpebral movements may depend on sympathetic activity, but the site of the generator is unclear. Such rhythmic pupillary and palpebral oscillation may be a feature of rapidly progressive CJD with predominant right hemispheric involvement. (c) 2009 Movement Disorder Society.

A semi-automated bacterial spore detection system (BSDS) was developed to detect biological threat agents (e.g., Bacillus anthracis) on-site. The system comprised an aerosol sampler, micro-fluidic chip-A (for spore germination and cell lysis), micro-fluidic chip-B (for extraction and detection of genomic DNA) and an analyzer. An aerosol with bacterial spores was first collected in the collection chamber of chip-A with a velocity of 300 l/min, and the chip-A was taken off from the aerosol sampler and loaded into the analyzer. Reagents packaged in the chip-A were sequentially applied into the chamber. The genomic DNA extract from spore lyzate was manually transferred from chip-A to chip-B and loaded into the analyzer. Genomic DNA in chip-B was first trapped on a glass bead column, washed with various reagents, and eluted to the detection chamber by sequential auto-dispensing. Isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN) with fluorescent measurement was adopted to amplify and detect target DNA. Bacillus subtilis was the stimulant of biological warfare agent in this experiment. Pretreatment conditions were optimized by examining bacterial target DNA recovery in the respective steps (aerosol collection, spore germination, cell lysis, and DNA extraction), by an off-chip experiment using a real-time polymerase chain reaction quantification method. Without the germination step, B. subtilis spores did not demonstrate amplification of target DNA. The detection of 10(4) spores was achieved within 2h throughout the micro-fluidic process.

The purpose of this study was to evaluate the clinical and pathological features in patients with progressive-type familial amyloidotic polyneuropathy (FAP) using autopsy and biopsy specimens. A proband is a 33-year-old man with FAP type I who developed motor, sensory and autonomic impairments with neuropathy, heart failure, and anorexia. Genetic findings of transthyretin (TTR) revealed G to A transition in codon 54 causing a rare mutation of TTR Lys54. He died of pneumonia and severe cardiac failure 4 years after onset. Autopsy showed heavy amyloid deposition in the heart, peripheral nerves, thyroid, skin, fat tissue, prostate and testis, moderate in the sympathetic nerve trunk, vagal nerve, celiac plexus, pelvic plexus, bladder, gastrointestinal tract, tongue, pancreas, lung, pituitary, blood vessel, gall bladder, adrenals and muscles, and free in the central nervous system, liver, kidney and spleen. Sural nerve biopsy in a sibling confirmed TTR amyloidosis immunohistochemically. Electronmicroscopic findings of amyloid fibrils were similar to that of FAP Met30. Immunoelectronmicroscopic findings indicated the relationship between amyloid fibrils or non-fibrillar structure and collagen fibers. The distribution of amyloid deposition, heavy in the heart and lacking in the kidney, is a characteristic feature and reflected severity of FAP with TTR Lys54.

In mammals and birds, most of the skeletal bones develop via endochondral ossification. Chondrocytes in the cartilaginous anlagen undergo processes of maturation such as hypertrophy, calcification and apoptosis. Concomitantly, osteoblasts are recruited to replace the cartilage scaffold gradually with bone matrix and become osteocytes in the trabecular bones. Throughout the successive development of bones, several gene products have been identified as being the components of the molecular mechanism regulating bone development. Transcription factor SOX9 plays essential roles during developmental steps from undifferentiated mesenchymal cells to proliferating chondrocytes, meanwhile, it inhibits transition of proliferating chondrocytes to hypertrophy. Other transcription factors RUNX2 and OSTERIX are critical in osteoblast differentiation, and RUNX2 is also essential for chondrocyte maturation such as hypertrophy and matrix mineralization. GDF5, a protein belonging to the transforming growth factor beta superfamily, is involved in joint formation and chondrogenesis. The limb skeleton of one of the ancestral tetrapod, anuran amphibians also develops through cartilaginous anlagen to bones, but their skeletogenesis has some unique characteristics compared with that of mammals and birds. Anuran amphibians develop and grow with less bone trabeculae and poor epiphyseal growth plates, and its endochondral ossification was found to be a delayed process. In order to address the characteristic skeletal development of anuran amphibians, we cloned Xenopus tropicalis RUNX2 (Xt-runx2), OSTERIX (Xt-osterix) and GDF5 (Xt-gdf5) homologue, and observed expression patterns together with Xt-sox9. In X. tropicalis limbs, histological observation and section in situ hybridization analysis suggest that Xt-SOX9 is involved in chondrogenesis, Xt-RUNX2 and Xt-OSTERIX are involved in osteogenesis, and Xt-GDF5 is involved in joint formation. In the cartilaginous anlagen, Xt-runx2 expression was found in perichondrium and immature chondrocytes as seen in other vertebrates. However, Xt-runx2 expression in enlarged chondrocytes was weak and dissimilar to common hypertrophic chondrocytes. These observations suggest that weak Xt-runx2 expression in maturing chondrocytes affects characteristic bone development in X. tropicalis long bones.

Conjunctival squamous cell carcinoma developed in a 51-year-old man 40 years after he had enucleation of his left eye because of an explosion injury. He had worn several ocular prostheses for more than 40 years. Recently he had noticed an increasing sanguineous conjunctival discharge, a foreign body sensation, and swelling of his left lower eyelid. Incisional biopsies of an underlying conjunctival mass revealed squamous cell carcinoma. His work history did not involve exposure to radiation, chemicals, or the sun. The authors concluded that squamous cell carcinoma in this case was caused by chronic irritation as the result of long-standing use of a poorly fitting ocular prosthesis.

Lateral ray polydactyly is the most common anomaly of the foot. However, when both the fifth and sixth toes are markedly shorter than a normal toe, reconstruction of a normal-length fifth toe has not been performed, because the toe rarely serves a functional purpose. The authors devised a new method whereby the fifth toe could be lengthened using the excess toe as a composite flap. Fifteen patients (15 toes) underwent our procedures and were evaluated in this study. The stepladder island flap is used. The decision as to which toe to use as the flap was made preoperatively based on nail size, and its morphologic and radiographic conditions. Stepladder incision lines were designed on the dorsal and plantar sides of the duplication. Care was taken to preserve the neurovascular bundle with the subcutaneous pedicle and osteotomies were made at the middle phalanx of the flap and the distal or middle phalanx of the recipient in almost all cases. The elevated composite toe flap was advanced and the bone was fixed rigidly for 1-2 months after the operation. During our follow-up periods, the reconstructed toes grew normally, and good aesthetic results had been achieved apart from the bulkiness of the reconstructed toes. There was no deficiency in circulation or function for walking, running, or shoe-fitting in this series.

Carbon monoxide (CO) is produced during the heme catabolism by heme oxygenase. In brain or blood vessels, CO functions as a neurotransmitter or an endothelial-derived relaxing factor. To verify whether crystallographically proposed CO-trapping sites of rat and cyanobacterial heme oxygenase-1 really work, heme catabolism by heme oxygenase-1 from rat and cyanobacterial Synechocystis sp. PCC 6803 has been scrutinized in the presence of 2-propanol. If 2-propanol occupies the trapping sites, formation of CO-bound verdoheme should be enhanced. Although effects of 2-propanol on the rat heme oxygenase-1 reaction were obscure, the reaction of cyanobacterial enzyme in the presence of NADPH/ferredoxin reductase/ferredoxin was apparently affected. Relative amount of CO-verdoheme versus CO-free verdoheme detected by optical absorption spectra increased as the equivalent of 2-propanol increased, thereby supporting indirectly that the hydrophobic cavity in cyanobacterial enzyme traps CO to reduce CO inhibition of verdoheme degradation.

We report a 75-year-old male with anaplastic carcinoma in an extrathyroid area. Thyroid remained unchanged. The patient is alive without incident of tumor recurrence at 3.5 years after total resection and at 5 years after initial symptom. The tumor developed between the sternocleidomastoid muscle and common carotid artery, and was completely separated from the thyroid. The tumor location was consistent with a branchial cyst. The tumor consisted of two parts; an upper solid tumor and a deep cystic tumor. The former showed anaplastic carcinoma with osteoclast-like giant cells. The latter was consistent with thyroid papillary carcinoma. The center was intermingled with these two carcinomas. Anaplastic carcinoma cells were positive for vimentin and papillary carcinoma cells were positive for keratin, thyroglobulin, and thyroid transcription factor-1. These results remain insufficient to find any conclusions concerning the tumor nature; either ectopic thyroid carcinoma arising from a branchial cyst or occult thyroid carcinoma metastasis. This is rare case in which thyroid anaplastic carcinoma transformed from papillary carcinoma in an extrathyroid area.

Cortactin is an F-actin binding protein that contributes to cytoskeleton remodelling. We identified five isoforms of mouse cortactin that differ in the number of tandem 37-amino acid repeats, named cortactin repeats. The transcription of minor isoforms with 4.5, 3.5 or 2.5 cortactin repeats was low in most adult tissues whereas an isoform with 4.5 cortactin repeats was highly transcribed in the adult brain. In accordance with the brain-specific upregulation of a minor isoform, a brain-specific novel 72-kDa cortactin protein was identified. Major isoforms with 6.5 or 5.5 cortactin repeats bound F-actin more robustly than minor isoforms in vitro. All isoforms were concentrated at cell-cell junction sites in epithelial cells. Deletion mutants lacking whole cortactin repeats did not bind F-actin and were not concentrated at cell-cell junction sites. Thus, the F-actin binding ability is mostly correlated with the number of cortactin repeats and is required for the cell-cell junctional localization.

Fenretinide, N-(4-hydroxyphenyl)retinamide (4-HPR), is a synthetic amide of all-trans-retinoic acid (RA), which inhibits cell growth, induces apoptosis, and is an antioxidant, and cancer chemopreventive and antiproliferative agent. These findings led us to investigate which structural component of 4-HPR contributes to these potent activities. Our approach was to examine 4-aminophenol (4-AP), p-methylaminophenol (p-MAP), and p-acetaminophen (p-AAP). It was found that vitamin E, 4-AP and p-MAP scavenge alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) radicals in a 1:2 ratio, in contrast to 4-HPR and p-AAP, for which 1:1 and 1:0.5 ratios were observed relative to DPPH radicals. However, RA was inactive. Lipid peroxidation in rat liver microsomes was reduced by compounds (RA > p-MAP = 4-HPR > 4-AP) in a dose-dependent manner, while p-AAP was inactive. In addition, both p-MAP and 4-HPR are potent inhibitors of cell growth and inducers of apoptosis in HL60 cells. p-MAP exhibits the same level of antiproliferative activity as 4-HPR against HL60R cells, which are a resistant clone against RA, and it inhibits the growth of various cancer cell lines (MCF-7, MCF-7/Adr(R), HepG2, and DU-145) to an extent greater than 4-AP and p-AAP, but is less potent than 4-HPR. Thus, although the antioxidant activity of p-MAP is more potent than that of 4-HPR, p-MAP is less potent than 4-HPR in anticancer activity. These results suggest that both the anticancer and antioxidative activities shown by 4-HPR are due to the structure of p-MAP. The retinoyl residue or long alkyl chain substituent attached to an aminophenol may be significant for anticancer properties.

Bromate, an inorganic oxyhalide disinfection by-product, is known to cause kidney damage, haemolysis and methaemoglobinemia. In potassium bromate (KBrO3)-treated mice (1.2 mmol/kg), elevation of methaemoglobin (MetHb) concentration in blood was observed simultaneously with an elevation of the NO concentration and attenuation of glutathione peroxidase (GPx) activity. Renal oxidative stress and kidney damage were also confirmed in the KBrO3-treated mice. A pre-administered GPx-mimic ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one) dose-dependently diminished the KBrO3-induced changes in MetHb concentration and GPx activity. Renal oxidative stress and kidney damage caused by the KBrO3 administration were also dose-dependently suppressed by ebselen. On the other hand, ebselen did not suppress the KBrO3-induced elevation of the NO concentration. KBrO3-induced methaemoglobinemia, renal oxidative stress and kidney damage, consequently, seemed to result from the attenuation of GPx activity. Besides, the enhancement of NO production was not likely to be a result but a cause for the KBrO3-induced attenuation of GPx activity. In in vitro experiments, oxidation of human oxyhaemoglobin (HbO2) to MetHb was observed in a reaction mixture containing HbO2 and an NO donor, NOC-7 (1-hydroxy-2-oxo-3-(N-methyl-3-aminopropyl)-3-methyl-1-triazene) or SIN-1 (3-(4-morpholinyl)sydnonimine), and this oxidation was inhibited by the NO scavenger carboxy-PTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide). However, no MetHb formation was observed in a reaction mixture containing HbO2 and KBrO3. These results suggest that KBrO3-induced methaemoglobinemia results from the reduction of GPx activity in blood by the KBrO3-induced increases in superoxide, NO and ONOO-.

Not only sulfur-oxidizing bacteria but also an acidophilic iron-oxidizing bacterium (or bacteria) were found in the corroded concrete from several sewerage systems in Japan. The surface pH of concrete test piece exposed to an atmosphere containing hydrogen sulfide of the concentrations more than 600 ppm in the systems was usually below 2 after a month. This was attributable to ability of the sulfur-oxidizing bacteria to grow in the thin water layer which contained hydrogen sulfide and covered the piece even when the surface pH of concrete was 12-13. When the sulfuroxidizing bacteria grew in the surface of concrete and produced sulfuric acid, the pH of the inner parts of concrete was lowered where the bacteria were hardly found. Probably, sulfuric acid formed by the bacteria in the surface parts penetrated into the inner parts. The different species of sulfur-oxidizing bacteria were found in different sewerage systems. The growth of the sulfur-oxidizing and acidophilic iron-oxidizing bacteria was completely inhibited by formates, especially by calcium formate of concentrations more than 50 mM. Calcium formate can protect concrete in sewerage systems from bacterial corrosion.

Human-placenta extract (PLx), has been widely used in clinical and cosmetic fields, and is known to possess marked antioxidant activity. We have attempted to identify the antioxidative components in PLx. Initially, we purified PLx using Sephadex G-10 column. The first eluted peak which had optical density at 280nm and exhibited approximately 20 per cent of antioxidant activity of the applied PLx, did not contain uracil, tyrosine, phenylalanine, or tryptophan which are the previously identified antioxidants from PLx. This fraction was further purified by reverse-phase high performance liquid chromatography. All eluted peaks containing antioxidant activity exhibited optical density at 280nm. Six separate fractions of eluent having antioxidant activity were analyzed by an amino acid sequencer, and each turned out to contain Glycine(G)-XY amino acid repeats, which appear to be derived from collagen. These results suggest that peptides produced from collagen are also antioxidative components of PLx.

An antioxidant was purified from human placenta extract (HPE) by using gel filtration, liquid-liquid extraction, silicagel column chromatography, and HPLC. The purified antioxidant was identified to be L-tryptophan (L-Trp). L-Trp showed higher inhibitory activity than mannitol and DMSO on the Fenton reaction-induced degradation of 2-deoxy-D-ribose. L-Trp also had much higher inhibitory activity on the cytochrome P-450-dependent lipid peroxidation than the previously identified antioxidants of HPE, L-phenylalanine, L-tyrosine and uracil. On the other hand, the inhibitory effect of L-Trp on the Fenton reaction-induced protein oxidation was smaller than that of uracil. These results suggest that L-Trp is a main antioxidant of HPE of which the effect is based on the suppression of lipid peroxidation in the oxidative stress status.

Intramedullary tuberculoma with syringomyelia is rare. We treated a woman with back pain and weakness of the left leg that had slowly progressed for more than 30 years. Radiologic evaluation demonstrated a crescent-shaped calcification at the level of the C6 vertebra, and syringomyelia from C7 to T9. Laminectomy and syringosubdural shunt placement were performed, and a tuberculoma was removed. Back pain resolved after the operation, and mobility was facilitated. We recommend surgery for intrathecal tuberculoma with syringomyelia even when the course has been prolonged with no active tuberculous lesion.

An actin filament binding protein cortactin was initially identified as a major phosphotyrosine-containing protein in v-Src-transformed chicken embryo fibroblast cells. The mouse, human, and Drosophila homologs were independently identified as a signaling molecule involved in a mitogenic response, as a product of a putative oncogene EMS1, and as a molecule interacting with a scaffolding protein ZO-1, respectively. In this report, we describe the cloning of the Drosophila cortactin gene, which consists of four exons and three introns, covering 3 kilobases in length. All exon-intron junctions are well matched with the GT/AG consensus sequence. S1 nuclease mapping revealed one major and several minor transcription start sites. The cytological location of the Drosophila cortactin gene is between chromosome segments 93B3 and 93B7.

Histologic examination of sural nerve of a 32-year-old man with an aggressive polyneuropathy associated with autonomic failure demonstrated amyloid deposition, and familial amyloidotic polyneuropathy (FAP) was diagnosed. Immunohistochemical staining showed transthyretin (TTR) staining of the amyloid deposits in nerve. Sequencing revealed G to A transition in the codon 54 causing TTR Lys 54. This is a new variant TTR associated with aggressive FAP.